AsCas12a

LOCUS      AsCas12a              12641 bp ds-DNA     circular SYN 28-SEP-2022
DEFINITION  Expresses FLAG tagged AsCas12a.
ACCESSION   .
VERSION     .
KEYWORDS    AsCas12a
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12641)
  AUTHORS   Escobar M, Li J, Patel A, Liu S, Xu Q, Hilton IB
  TITLE     Quantification of Genome Editing and Transcriptional Control 
            Capabilities Reveals Hierarchies among Diverse CRISPR/Cas Systems in
            Human Cells
  JOURNAL   ACS Synth. Biol. 2022
REFERENCE   2  (bases 1 to 12641)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 28, 2022 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..12641
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..199
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     154..174
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             217..397
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    444..569
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1062..1295
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1480..1524
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    1822..1939
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_RNA        2071..2146
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2208..2419
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             2444..6364
                     /codon_start=1
                     /product="CRISPR-associated protein from Acidaminococcus 
                     sp. BV3L6 (Zetsche et al., 2015)"
                     /label=AsCpf1
                     /translation="MTQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARND
                     HYKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRN
                     AIHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFD
                     KFTTYFSGFYENRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHF
                     ENVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVL
                     NLAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLL
                     RNENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTG
                     KITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTL
                     KKQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKA
                     RNYATKKPYSVEKFKLNFQMPTLASGWDVNKEKNNGAILFVKNGLYYLGIMPKQKGRYK
                     ALSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEP
                     LEITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSID
                     LSSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKG
                     HHGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLK
                     DQKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFF
                     HVPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVIDSTGKILEQR
                     SLNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAV
                     VVLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQ
                     FTSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYD
                     VKTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIEN
                     HRFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQM
                     RNSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKES
                     KDLKLQNGISNQDWLAYIQELRN"
     regulatory      6084..6093
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             6365..6412
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6413..6436
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             6446..6502
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             6503..7099
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     6996..7016
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7115..7703
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7168..7188)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7586..7597)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7775..8008
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(8034..8051)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8040..8264
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8310..8738
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8397..8416)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8607..8628
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8747..8767)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8752..9081
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8932..9067
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8994..9013
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9129..9176
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9195..9569
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    9699..9820
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9736..9755)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9790..9809
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9869..9885)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9869..9885)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9882..9904)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9893..9909
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(9917..9947)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9962..9983
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10100..10117)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10271..10859)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10351..10370)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(11030..11890)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     11653..11672
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11891..11995)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(12070..12089)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        12261..12640
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
       61 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      121 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      181 gggaggtcta tataagcagc gcgttttgcc tgtactgggt ctctctggtt agaccagatc
      241 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
      301 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
      361 ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga
      421 aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca
      481 cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct
      541 agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat
      601 gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat
      661 gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag
      721 gctgtagaca aatactggga cagctacaac catcccttca gacaggatca gaagaactta
      781 gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag
      841 acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac
      901 agcaagcggc cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt
      961 gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca
     1021 aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg
     1081 ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc
     1141 agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg
     1201 caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg
     1261 gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa
     1321 ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag
     1381 atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta
     1441 atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg
     1501 gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat
     1561 ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta
     1621 ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc
     1681 ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac
     1741 agagacagat ccattcgatt agtgaacgga tcggcactgc gtgcgccaat tctgcagaca
     1801 aatggcagta ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg
     1861 ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat
     1921 tacaaaaatt caaaattttc gggtttatta cagggacagc agagatccag tttggttaat
     1981 taaggtaccg atatcactag tgttccatta actgcgctaa aaataatttt taaatctttt
     2041 ttagcttctt gctctttttt gtacgtctct gttttagagc tagaaatagc aagttaaaat
     2101 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttgaattcgc
     2161 tagctaggtc ttgaaaggag tgggaattgg ctccggtgcc cgtcagtggg cagagcgcac
     2221 atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgatccg gtgcctagag
     2281 aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga
     2341 gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg
     2401 gtttgccgcc agaacacagg accggttcta gagcgctgcc accatgacac agttcgaggg
     2461 ctttaccaac ctgtatcagg tgagcaagac actgcggttt gagctgatcc cacagggcaa
     2521 gaccctgaag cacatccagg agcagggctt catcgaggag gacaaggccc gcaatgatca
     2581 ctacaaggag ctgaagccca tcatcgatcg gatctacaag acctatgccg accagtgcct
     2641 gcagctggtg cagctggatt gggagaacct gagcgccgcc atcgactcct atagaaagga
     2701 gaaaaccgag gagacaagga acgccctgat cgaggagcag gccacatatc gcaatgccat
     2761 ccacgactac ttcatcggcc ggacagacaa cctgaccgat gccatcaata agagacacgc
     2821 cgagatctac aagggcctgt tcaaggccga gctgtttaat ggcaaggtgc tgaagcagct
     2881 gggcaccgtg accacaaccg agcacgagaa cgccctgctg cggagcttcg acaagtttac
     2941 aacctacttc tccggctttt atgagaacag gaagaacgtg ttcagcgccg aggatatcag
     3001 cacagccatc ccacaccgca tcgtgcagga caacttcccc aagtttaagg agaattgtca
     3061 catcttcaca cgcctgatca ccgccgtgcc cagcctgcgg gagcactttg agaacgtgaa
     3121 gaaggccatc ggcatcttcg tgagcacctc catcgaggag gtgttttcct tcccttttta
     3181 taaccagctg ctgacacaga cccagatcga cctgtataac cagctgctgg gaggaatctc
     3241 tcgggaggca ggcaccgaga agatcaaggg cctgaacgag gtgctgaatc tggccatcca
     3301 gaagaatgat gagacagccc acatcatcgc ctccctgcca cacagattca tccccctgtt
     3361 taagcagatc ctgtccgata ggaacaccct gtctttcatc ctggaggagt ttaagagcga
     3421 cgaggaagtg atccagtcct tctgcaagta caagacactg ctgagaaacg agaacgtgct
     3481 ggagacagcc gaggccctgt ttaacgagct gaacagcatc gacctgacac acatcttcat
     3541 cagccacaag aagctggaga caatcagcag cgccctgtgc gaccactggg atacactgag
     3601 gaatgccctg tatgagcgga gaatctccga gctgacaggc aagatcacca agtctgccaa
     3661 ggagaaggtg cagcgcagcc tgaagcacga ggatatcaac ctgcaggaga tcatctctgc
     3721 cgcaggcaag gagctgagcg aggccttcaa gcagaaaacc agcgagatcc tgtcccacgc
     3781 acacgccgcc ctggatcagc cactgcctac aaccctgaag aagcaggagg agaaggagat
     3841 cctgaagtct cagctggaca gcctgctggg cctgtaccac ctgctggact ggtttgccgt
     3901 ggatgagtcc aacgaggtgg accccgagtt ctctgcccgg ctgaccggca tcaagctgga
     3961 gatggagcct tctctgagct tctacaacaa ggccagaaat tatgccacca agaagcccta
     4021 ctccgtggag aagttcaagc tgaactttca gatgcctaca ctggcctctg gctgggacgt
     4081 gaataaggag aagaacaatg gcgccatcct gtttgtgaag aacggcctgt actatctggg
     4141 catcatgcca aagcagaagg gcaggtataa ggccctgagc ttcgagccca cagagaaaac
     4201 cagcgagggc tttgataaga tgtactatga ctacttccct gatgccgcca agatgatccc
     4261 aaagtgcagc acccagctga aggccgtgac agcccacttt cagacccaca caacccccat
     4321 cctgctgtcc aacaatttca tcgagcctct ggagatcaca aaggagatct acgacctgaa
     4381 caatcctgag aaggagccaa agaagtttca gacagcctac gccaagaaaa ccggcgacca
     4441 gaagggctac agagaggccc tgtgcaagtg gatcgacttc acaagggatt ttctgtccaa
     4501 gtataccaag acaacctcta tcgatctgtc tagcctgcgg ccatcctctc agtataagga
     4561 cctgggcgag tactatgccg agctgaatcc cctgctgtac cacatcagct tccagagaat
     4621 cgccgagaag gagatcatgg atgccgtgga gacaggcaag ctgtacctgt tccagatcta
     4681 taacaaggac tttgccaagg gccaccacgg caagcctaat ctgcacacac tgtattggac
     4741 cggcctgttt tctccagaga acctggccaa gacaagcatc aagctgaatg gccaggccga
     4801 gctgttctac cgccctaagt ccaggatgaa gaggatggca caccggctgg gagagaagat
     4861 gctgaacaag aagctgaagg atcagaaaac cccaatcccc gacaccctgt accaggagct
     4921 gtacgactat gtgaatcaca gactgtccca cgacctgtct gatgaggcca gggccctgct
     4981 gcccaacgtg atcaccaagg aggtgtctca cgagatcatc aaggataggc gctttaccag
     5041 cgacaagttc tttttccacg tgcctatcac actgaactat caggccgcca attccccatc
     5101 taagttcaac cagagggtga atgcctacct gaaggagcac cccgagacac ctatcatcgg
     5161 catcgatcgg ggcgagagaa acctgatcta tatcacagtg atcgactcca ccggcaagat
     5221 cctggagcag cggagcctga acaccatcca gcagtttgat taccagaaga agctggacaa
     5281 cagggagaag gagagggtgg cagcaaggca ggcctggtct gtggtgggca caatcaagga
     5341 tctgaagcag ggctatctga gccaggtcat ccacgagatc gtggacctga tgatccacta
     5401 ccaggccgtg gtggtgctgg agaacctgaa tttcggcttt aagagcaaga ggaccggcat
     5461 cgccgagaag gccgtgtacc agcagttcga gaagatgctg atcgataagc tgaattgcct
     5521 ggtgctgaag gactatccag cagagaaagt gggaggcgtg ctgaacccat accagctgac
     5581 agaccagttc acctcctttg ccaagatggg cacccagtct ggcttcctgt tttacgtgcc
     5641 tgccccatat acatctaaga tcgatcccct gaccggcttc gtggacccct tcgtgtggaa
     5701 aaccatcaag aatcacgaga gccgcaagca cttcctggag ggcttcgact ttctgcacta
     5761 cgacgtgaaa accggcgact tcatcctgca ctttaagatg aacagaaatc tgtccttcca
     5821 gaggggcctg cccggcttta tgcctgcatg ggatatcgtg ttcgagaaga acgagacaca
     5881 gtttgacgcc aagggcaccc ctttcatcgc cggcaagaga atcgtgccag tgatcgagaa
     5941 tcacagattc accggcagat accgggacct gtatcctgcc aacgagctga tcgccctgct
     6001 ggaggagaag ggcatcgtgt tcagggatgg ctccaacatc ctgccaaagc tgctggagaa
     6061 tgacgattct cacgccatcg acaccatggt ggccctgatc cgcagcgtgc tgcagatgcg
     6121 gaactccaat gccgccacag gcgaggacta tatcaacagc cccgtgcgcg atctgaatgg
     6181 cgtgtgcttc gactcccggt ttcagaaccc agagtggccc atggacgccg atgccaatgg
     6241 cgcctaccac atcgccctga agggccagct gctgctgaat cacctgaagg agagcaagga
     6301 tctgaagctg cagaacggca tctccaatca ggactggctg gcctacatcc aggagctgcg
     6361 caacaagcga cctgccgcca caaagaaggc tggacaggct aagaagaaga aagattacaa
     6421 agacgatgac gataagggat ccggcgcaac aaacttctct ctgctgaaac aagccggaga
     6481 tgtcgaagag aatcctggac cgaccgagta caagcccacg gtgcgcctcg ccacccgcga
     6541 cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg
     6601 ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct
     6661 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc
     6721 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg
     6781 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc
     6841 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggagtct cgcccgacca
     6901 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc
     6961 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct
     7021 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac
     7081 ccgcaagccc ggtgcctgaa cgcgttaagt cgacaatcaa cctctggatt acaaaatttg
     7141 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     7201 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     7261 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     7321 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     7381 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     7441 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     7501 gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg
     7561 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     7621 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     7681 ctccctttgg gccgcctccc cgcgtcgact ttaagaccaa tgacttacaa ggcagctgta
     7741 gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga
     7801 agacaagatc tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg
     7861 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     7921 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     7981 ttttagtcag tgtggaaaat ctctagcagg gcccgtttaa acccgctgat cagcctcgac
     8041 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     8101 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     8161 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     8221 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     8281 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     8341 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     8401 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     8461 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     8521 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     8581 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     8641 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     8701 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     8761 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     8821 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     8881 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     8941 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     9001 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     9061 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     9121 cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg
     9181 tgaggaacta aaccatggcc aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg
     9241 tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg
     9301 acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg gtccaggacc
     9361 aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg cggcctggac gagctgtacg
     9421 ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc ctccgggccg gccatgaccg
     9481 agatcggcga gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg
     9541 tgcacttcgt ggccgaggag caggactgac acgtgctacg agatttcgat tccaccgccg
     9601 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     9661 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     9721 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     9781 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     9841 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     9901 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     9961 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
    10021 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
    10081 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
    10141 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
    10201 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
    10261 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
    10321 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
    10381 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
    10441 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
    10501 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
    10561 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
    10621 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
    10681 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
    10741 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
    10801 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
    10861 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
    10921 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
    10981 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
    11041 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
    11101 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
    11161 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
    11221 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
    11281 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
    11341 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
    11401 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
    11461 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
    11521 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
    11581 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
    11641 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
    11701 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
    11761 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
    11821 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
    11881 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
    11941 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
    12001 ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg
    12061 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg
    12121 tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt
    12181 gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt
    12241 acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac
    12301 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
    12361 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
    12421 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
    12481 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
    12541 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
    12601 ttggcagtac atctacgtat tagtcatcgc tattaccatg g
//