Lenti-AsCpf1-Blast

LOCUS       Lenti-AsCpf1-Blast               12670 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses human codon-optimized AsCpf1 protein and blasticidin 
            resistance from EFS promoter. Lentiviral backbone..
ACCESSION   .
VERSION     .
KEYWORDS    Lenti-AsCpf1-Blast
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12670)
  AUTHORS   Kim HK, Song M, Lee J, Menon AV, Jung S, Kang YM, Choi JW, Woo E, 
            Koh HC, Nam JW, Kim H
  TITLE     In vivo high-throughput profiling of CRISPR-Cpf1 activity.
  JOURNAL   Nat Methods. 2016 Dec 19. doi: 10.1038/nmeth.4104.
  PUBMED    27992409
REFERENCE   2  (bases 1 to 12670)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..12670
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /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    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2650..2861
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             2903..6823
                     /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      6543..6552
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             6824..6871
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6881..6937
                     /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             6938..7336
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    7361..7949
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7414..7434)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7832..7843)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(7952..7968)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7953..7969)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             8474..8654
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(8680..8697)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8686..8910
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8956..9384
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9043..9062)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9253..9274
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9393..9413)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9398..9728
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9580..9714
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9641..9660
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9776..9823
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9842..10216
                     /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    10346..10467
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10383..10402)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10437..10456
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10516..10532)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10516..10532)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10529..10551)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10540..10556
                     /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(10564..10594)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10609..10630
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10747..10764)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10918..11506)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10998..11017)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(11677..12537)
                     /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     12300..12319
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12538..12642)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttaatta gctagctagg tcttgaaagg agtgggaatt ggctccggtg
     2641 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
     2701 gcaattgatc cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
     2761 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
     2821 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggaccggttc tagcgtttaa
     2881 acttaagctt ggtaccgcca ccatgacaca gttcgagggc tttaccaacc tgtatcaggt
     2941 gagcaagaca ctgcggtttg agctgatccc acagggcaag accctgaagc acatccagga
     3001 gcagggcttc atcgaggagg acaaggcccg caatgatcac tacaaggagc tgaagcccat
     3061 catcgatcgg atctacaaga cctatgccga ccagtgcctg cagctggtgc agctggattg
     3121 ggagaacctg agcgccgcca tcgactccta tagaaaggag aaaaccgagg agacaaggaa
     3181 cgccctgatc gaggagcagg ccacatatcg caatgccatc cacgactact tcatcggccg
     3241 gacagacaac ctgaccgatg ccatcaataa gagacacgcc gagatctaca agggcctgtt
     3301 caaggccgag ctgtttaatg gcaaggtgct gaagcagctg ggcaccgtga ccacaaccga
     3361 gcacgagaac gccctgctgc ggagcttcga caagtttaca acctacttct ccggctttta
     3421 tgagaacagg aagaacgtgt tcagcgccga ggatatcagc acagccatcc cacaccgcat
     3481 cgtgcaggac aacttcccca agtttaagga gaattgtcac atcttcacac gcctgatcac
     3541 cgccgtgccc agcctgcggg agcactttga gaacgtgaag aaggccatcg gcatcttcgt
     3601 gagcacctcc atcgaggagg tgttttcctt ccctttttat aaccagctgc tgacacagac
     3661 ccagatcgac ctgtataacc agctgctggg aggaatctct cgggaggcag gcaccgagaa
     3721 gatcaagggc ctgaacgagg tgctgaatct ggccatccag aagaatgatg agacagccca
     3781 catcatcgcc tccctgccac acagattcat ccccctgttt aagcagatcc tgtccgatag
     3841 gaacaccctg tctttcatcc tggaggagtt taagagcgac gaggaagtga tccagtcctt
     3901 ctgcaagtac aagacactgc tgagaaacga gaacgtgctg gagacagccg aggccctgtt
     3961 taacgagctg aacagcatcg acctgacaca catcttcatc agccacaaga agctggagac
     4021 aatcagcagc gccctgtgcg accactggga tacactgagg aatgccctgt atgagcggag
     4081 aatctccgag ctgacaggca agatcaccaa gtctgccaag gagaaggtgc agcgcagcct
     4141 gaagcacgag gatatcaacc tgcaggagat catctctgcc gcaggcaagg agctgagcga
     4201 ggccttcaag cagaaaacca gcgagatcct gtcccacgca cacgccgccc tggatcagcc
     4261 actgcctaca accctgaaga agcaggagga gaaggagatc ctgaagtctc agctggacag
     4321 cctgctgggc ctgtaccacc tgctggactg gtttgccgtg gatgagtcca acgaggtgga
     4381 ccccgagttc tctgcccggc tgaccggcat caagctggag atggagcctt ctctgagctt
     4441 ctacaacaag gccagaaatt atgccaccaa gaagccctac tccgtggaga agttcaagct
     4501 gaactttcag atgcctacac tggcctctgg ctgggacgtg aataaggaga agaacaatgg
     4561 cgccatcctg tttgtgaaga acggcctgta ctatctgggc atcatgccaa agcagaaggg
     4621 caggtataag gccctgagct tcgagcccac agagaaaacc agcgagggct ttgataagat
     4681 gtactatgac tacttccctg atgccgccaa gatgatccca aagtgcagca cccagctgaa
     4741 ggccgtgaca gcccactttc agacccacac aacccccatc ctgctgtcca acaatttcat
     4801 cgagcctctg gagatcacaa aggagatcta cgacctgaac aatcctgaga aggagccaaa
     4861 gaagtttcag acagcctacg ccaagaaaac cggcgaccag aagggctaca gagaggccct
     4921 gtgcaagtgg atcgacttca caagggattt tctgtccaag tataccaaga caacctctat
     4981 cgatctgtct agcctgcggc catcctctca gtataaggac ctgggcgagt actatgccga
     5041 gctgaatccc ctgctgtacc acatcagctt ccagagaatc gccgagaagg agatcatgga
     5101 tgccgtggag acaggcaagc tgtacctgtt ccagatctat aacaaggact ttgccaaggg
     5161 ccaccacggc aagcctaatc tgcacacact gtattggacc ggcctgtttt ctccagagaa
     5221 cctggccaag acaagcatca agctgaatgg ccaggccgag ctgttctacc gccctaagtc
     5281 caggatgaag aggatggcac accggctggg agagaagatg ctgaacaaga agctgaagga
     5341 tcagaaaacc ccaatccccg acaccctgta ccaggagctg tacgactatg tgaatcacag
     5401 actgtcccac gacctgtctg atgaggccag ggccctgctg cccaacgtga tcaccaagga
     5461 ggtgtctcac gagatcatca aggataggcg ctttaccagc gacaagttct ttttccacgt
     5521 gcctatcaca ctgaactatc aggccgccaa ttccccatct aagttcaacc agagggtgaa
     5581 tgcctacctg aaggagcacc ccgagacacc tatcatcggc atcgatcggg gcgagagaaa
     5641 cctgatctat atcacagtga tcgactccac cggcaagatc ctggagcagc ggagcctgaa
     5701 caccatccag cagtttgatt accagaagaa gctggacaac agggagaagg agagggtggc
     5761 agcaaggcag gcctggtctg tggtgggcac aatcaaggat ctgaagcagg gctatctgag
     5821 ccaggtcatc cacgagatcg tggacctgat gatccactac caggccgtgg tggtgctgga
     5881 gaacctgaat ttcggcttta agagcaagag gaccggcatc gccgagaagg ccgtgtacca
     5941 gcagttcgag aagatgctga tcgataagct gaattgcctg gtgctgaagg actatccagc
     6001 agagaaagtg ggaggcgtgc tgaacccata ccagctgaca gaccagttca cctcctttgc
     6061 caagatgggc acccagtctg gcttcctgtt ttacgtgcct gccccatata catctaagat
     6121 cgatcccctg accggcttcg tggacccctt cgtgtggaaa accatcaaga atcacgagag
     6181 ccgcaagcac ttcctggagg gcttcgactt tctgcactac gacgtgaaaa ccggcgactt
     6241 catcctgcac tttaagatga acagaaatct gtccttccag aggggcctgc ccggctttat
     6301 gcctgcatgg gatatcgtgt tcgagaagaa cgagacacag tttgacgcca agggcacccc
     6361 tttcatcgcc ggcaagagaa tcgtgccagt gatcgagaat cacagattca ccggcagata
     6421 ccgggacctg tatcctgcca acgagctgat cgccctgctg gaggagaagg gcatcgtgtt
     6481 cagggatggc tccaacatcc tgccaaagct gctggagaat gacgattctc acgccatcga
     6541 caccatggtg gccctgatcc gcagcgtgct gcagatgcgg aactccaatg ccgccacagg
     6601 cgaggactat atcaacagcc ccgtgcgcga tctgaatggc gtgtgcttcg actcccggtt
     6661 tcagaaccca gagtggccca tggacgccga tgccaatggc gcctaccaca tcgccctgaa
     6721 gggccagctg ctgctgaatc acctgaagga gagcaaggat ctgaagctgc agaacggcat
     6781 ctccaatcag gactggctgg cctacatcca ggagctgcgc aacaaaaggc cggcggccac
     6841 gaaaaaggcc ggccaggcaa aaaagaaaaa gggatccggc gcaacaaact tctctctgct
     6901 gaaacaagcc ggagatgtcg aagagaatcc tggaccgatg gccaagcctt tgtctcaaga
     6961 agaatccacc ctcattgaaa gagcaacggc tacaatcaac agcatcccca tctctgaaga
     7021 ctacagcgtc gccagcgcag ctctctctag cgacggccgc atcttcactg gtgtcaatgt
     7081 atatcatttt actgggggac cttgtgcaga actcgtggtg ctgggcactg ctgctgctgc
     7141 ggcagctggc aacctgactt gtatcgtcgc gatcggaaat gagaacaggg gcatcttgag
     7201 cccctgcgga cggtgccgac aggtgcttct cgatctgcat cctgggatca aagccatagt
     7261 gaaggacagt gatggacagc cgacggcagt tgggattcgt gaattgctgc cctctggtta
     7321 tgtgtgggag ggctaagaat tcgatatcaa gcttatcggt aatcaacctc tggattacaa
     7381 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     7441 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     7501 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     7561 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     7621 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     7681 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     7741 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat
     7801 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     7861 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     7921 tcggatctcc ctttgggccg cctccccgca tcgataccgt cgacctcgag acctagaaaa
     7981 acatggagca atcacaagta gcaatacagc agctaccaat gctgattgtg cctggctaga
     8041 agcacaagag gaggaggagg tgggttttcc agtcacacct caggtacctt taagaccaat
     8101 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg
     8161 gctaattcac tcccaacgaa gacaagatat ccttgatctg tggatctacc acacacaagg
     8221 ctacttccct gattggcaga actacacacc agggccaggg atcagatatc cactgacctt
     8281 tggatggtgc tacaagctag taccagttga gcaagagaag gtagaagaag ccaatgaagg
     8341 agagaacacc cgcttgttac accctgtgag cctgcatggg atggatgacc cggagagaga
     8401 agtattagag tggaggtttg acagccgcct agcatttcat cacatggccc gagagctgca
     8461 tccggactgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     8521 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     8581 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     8641 gaaaatctct agcagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
     8701 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     8761 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     8821 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     8881 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     8941 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     9001 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     9061 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     9121 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     9181 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     9241 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     9301 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     9361 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     9421 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     9481 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     9541 ttagtcagca accatagtcc cgcccctaac tccgcccagt cccgccccta actccgccca
     9601 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     9661 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     9721 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcag cacgtgttga
     9781 caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac
     9841 catggccaag ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt
     9901 cgagttctgg accgaccggc tcgggttctc ccgggacttc gtggaggacg acttcgccgg
     9961 tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc caggaccagg tggtgccgga
    10021 caacaccctg gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga
    10081 ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc atgaccgaga tcggcgagca
    10141 gccgtggggg cgggagttcg ccctgcgcga cccggccggc aactgcgtgc acttcgtggc
    10201 cgaggagcag gactgacacg tgctacgaga tttcgattcc accgccgcct tctatgaaag
    10261 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
    10321 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
    10381 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
    10441 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
    10501 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
    10561 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
    10621 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
    10681 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
    10741 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
    10801 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
    10861 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
    10921 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
    10981 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
    11041 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
    11101 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
    11161 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
    11221 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
    11281 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
    11341 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
    11401 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
    11461 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
    11521 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
    11581 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
    11641 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
    11701 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
    11761 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
    11821 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
    11881 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
    11941 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
    12001 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
    12061 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
    12121 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
    12181 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
    12241 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
    12301 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
    12361 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
    12421 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
    12481 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
    12541 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
    12601 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
    12661 gccacctgac
//