opCas12a-T8N(pZZ34, TAT-6MycNLS-opCas12a-2SV40NLS-sfGFP)

LOCUS       opCas12a-T8N(pZZ34,TAT-6MycNLS-opCas12a-2SV40NLS-sfGFP)              11057 bp ds-DNA     circular SYN 28-APR-2023
DEFINITION  Expression of cell permeable opAsCas12a in E. Coli..
ACCESSION   .
VERSION     .
KEYWORDS    opCas12a-T8N (pZZ34, TAT-6MycNLS-opCas12a-2SV40NLS-sfGFP)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11057)
  AUTHORS   Zhang Z, Baxter AE, Ren D, Qin K, Chen Z, Collins SM, Huang H, Komar
            CA, Bailer PF, Parker JB, Blobel GA, Kohli RM, Wherry EJ, Berger SL,
            Shi J
  TITLE     Efficient engineering of human and mouse primary cells using 
            peptide-assisted genome editing
  JOURNAL   Nature Biotechnology. 2023 April 24
REFERENCE   2  (bases 1 to 11057)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 28, 2023 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..11057
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             complement(1328..2410)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     2372..2391
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2411..2488)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2694..2713
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2797..2816
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2797..2815
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2816..2840
                     /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)."
     RBS             2855..2877
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2896..2913
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2923..2940
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2953..3036
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="WSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     CDS             3037..3327
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
                     RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIG"
     CDS             3379..3405
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             3415..3441
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             3451..3477
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             3487..3513
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             3523..3549
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             3559..3585
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             3613..7530
                     /codon_start=1
                     /product="CRISPR-associated protein from Acidaminococcus 
                     sp. BV3L6 (Zetsche et al., 2015)"
                     /label=AsCpf1
                     /translation="TQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARNDH
                     YKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRNA
                     IHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFDK
                     FTTYFSGFYRNRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHFE
                     NVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVLN
                     LAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLLR
                     NENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTGK
                     ITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTLK
                     KQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKAR
                     NYATKKPYSVEKFKLNFQMPTLARGWDVNKEKNNGAILFVKNGLYYLGIMPKQKGRYKA
                     LSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEPL
                     EITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSIDL
                     SSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKGH
                     HGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLKD
                     QKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFFH
                     VPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVIDSTGKILEQRS
                     LNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAVV
                     VLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQF
                     TSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYDV
                     KTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIENH
                     RFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQMR
                     NSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKESK
                     DLKLQNGISNQDWLAYIQELRN"
     regulatory      7250..7259
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             7537..7557
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7564..7584
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(8354..8372)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      8368..8415
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(8590..8618)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(8645..8663)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8731..8835
                     /gene="bla"
                     /label=AmpR promoter
     CDS             8836..9696
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(9054..9073)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9867..10455
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10356..10375
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10609..10626
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    10641..10780
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(10706..10725)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     10866..10888
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(10882..16)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga
       61 taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt
      121 tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa
      181 tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc
      241 agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt
      301 ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg
      361 ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag
      421 taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc
      481 gtggccagga cccaacgctg cccgagatgc gccgcgtgcg gctgctggag atggcggacg
      541 cgatggatat gttctgccaa gggttggttt gcgcattcac agttctccgc aagaattgat
      601 tggctccaat tcttggagtg gtgaatccgt tagcgaggtg ccgccggctt ccattcaggt
      661 cgaggtggcc cggctccatg caccgcgacg caacgcgggg aggcagacaa ggtatagggc
      721 ggcgcctaca atccatgcca acccgttcca tgtgctcgcc gaggcggcat aaatcgccgt
      781 gacgatcagc ggtccaatga tcgaagttag gctggtaaga gccgcgagcg atccttgaag
      841 ctgtccctga tggtcgtcat ctacctgcct ggacagcatg gcctgcaacg cgggcatccc
      901 gatgccgccg gaagcgagaa gaatcataat ggggaaggcc atccagcctc gcgtcgcgaa
      961 cgccagcaag acgtagccca gcgcgtcggc cgccatgccg gcgataatgg cctgcttctc
     1021 gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc
     1081 gaataccgca agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa
     1141 aatgacccag agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat
     1201 aagtgcggcg acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc
     1261 tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc
     1321 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     1381 cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca
     1441 ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca
     1501 agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg
     1561 ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagata tccgcaccaa
     1621 cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa
     1681 ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg
     1741 acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat
     1801 atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca
     1861 gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt
     1921 catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg
     1981 gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa
     2041 tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga
     2101 cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt
     2161 taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa
     2221 tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct
     2281 ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca
     2341 ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta
     2401 ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc
     2461 gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc
     2521 tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat
     2581 ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc
     2641 acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg
     2701 tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt
     2761 ccggcgtaga ggatcgagat ctcgatcccg cgaaattaat acgactcact ataggggaat
     2821 tgtgagcgga taacaattcc cctctagaaa taattttgtt taactttaag aaggagatat
     2881 accatgggca gcagccatca tcatcatcat cacagcagcg gcctggtgcc gcgcggcagc
     2941 catatggcta gctggagcca tccgcagttt gaaaaaggtg gtggtagcgg tggtggttca
     3001 ggtggtagtg catggtcaca ccctcagttt gagaaaatgt cggactcaga agtcaatcaa
     3061 gaagctaagc cagaggtcaa gccagaagtc aagcctgaga ctcacatcaa tttaaaggtg
     3121 tccgatggat cttcagagat cttcttcaag atcaaaaaga ccactccttt aagaaggctg
     3181 atggaagcgt tcgctaaaag acagggtaag gaaatggact ccttaagatt cttgtacgac
     3241 ggtattagaa tccaagctga tcagacccct gaagatttgg acatggagga taacgatatt
     3301 attgaggctc acagagaaca gattggtgga tccggccgta agaaacgccg ccagcgtcgc
     3361 cgtccgcaag gcggtagccc ggctgcaaag cgcgtcaaat tggatggtgg gtcacctgcg
     3421 gctaagcgag tcaaattaga cggaggatcg cccgccgcga agcgtgtaaa gctggatggg
     3481 ggtagcccag cggcaaaaag ggtgaaacta gacggcggct ccccggcagc caagagagtc
     3541 aaactcgacg ggggtagccc tgctgctaaa cgtgttaagc ttgatggtat ccacggagtc
     3601 ccagcagcta ccacacagtt cgagggcttt accaacctgt atcaggtgag caagacactg
     3661 cggtttgagc tgatcccaca gggcaagacc ctgaagcaca tccaggagca gggcttcatc
     3721 gaggaggaca aggcccgcaa tgatcactac aaggagctga agcccatcat cgatcggatc
     3781 tacaagacct atgccgacca gtgcctgcag ctggtgcagc tggattggga gaacctgagc
     3841 gccgccatcg actcctatag aaaggagaaa accgaggaga caaggaacgc cctgatcgag
     3901 gagcaggcca catatcgcaa tgccatccac gactacttca tcggccggac agacaacctg
     3961 accgatgcca tcaataagag acacgccgag atctacaagg gcctgttcaa ggccgagctg
     4021 tttaatggca aggtgctgaa gcagctgggc accgtgacca caaccgagca cgagaacgcc
     4081 ctgctgcgga gcttcgacaa gtttacaacc tacttctccg gcttttatcg gaacaggaag
     4141 aacgtgttca gcgccgagga tatcagcaca gccatcccac accgcatcgt gcaggacaac
     4201 ttccccaagt ttaaggagaa ttgtcacatc ttcacacgcc tgatcaccgc cgtgcccagc
     4261 ctgcgggagc actttgagaa cgtgaagaag gccatcggca tcttcgtgag cacctccatc
     4321 gaggaggtgt tttccttccc tttttataac cagctgctga cacagaccca gatcgacctg
     4381 tataaccagc tgctgggagg aatctctcgg gaggcaggca ccgagaagat caagggcctg
     4441 aacgaggtgc tgaatctggc catccagaag aatgatgaga cagcccacat catcgcctcc
     4501 ctgccacaca gattcatccc cctgtttaag cagatcctgt ccgataggaa caccctgtct
     4561 ttcatcctgg aggagtttaa gagcgacgag gaagtgatcc agtccttctg caagtacaag
     4621 acactgctga gaaacgagaa cgtgctggag acagccgagg ccctgtttaa cgagctgaac
     4681 agcatcgacc tgacacacat cttcatcagc cacaagaagc tggagacaat cagcagcgcc
     4741 ctgtgcgacc actgggatac actgaggaat gccctgtatg agcggagaat ctccgagctg
     4801 acaggcaaga tcaccaagtc tgccaaggag aaggtgcagc gcagcctgaa gcacgaggat
     4861 atcaacctgc aggagatcat ctctgccgca ggcaaggagc tgagcgaggc cttcaagcag
     4921 aaaaccagcg agatcctgtc ccacgcacac gccgccctgg atcagccact gcctacaacc
     4981 ctgaagaagc aggaggagaa ggagatcctg aagtctcagc tggacagcct gctgggcctg
     5041 taccacctgc tggactggtt tgccgtggat gagtccaacg aggtggaccc cgagttctct
     5101 gcccggctga ccggcatcaa gctggagatg gagccttctc tgagcttcta caacaaggcc
     5161 agaaattatg ccaccaagaa gccctactcc gtggagaagt tcaagctgaa ctttcagatg
     5221 cctacactgg cccgtggctg ggacgtgaat aaggagaaga acaatggcgc catcctgttt
     5281 gtgaagaacg gcctgtacta tctgggcatc atgccaaagc agaagggcag gtataaggcc
     5341 ctgagcttcg agcccacaga gaaaaccagc gagggctttg ataagatgta ctatgactac
     5401 ttccctgatg ccgccaagat gatcccaaag tgcagcaccc agctgaaggc cgtgacagcc
     5461 cactttcaga cccacacaac ccccatcctg ctgtccaaca atttcatcga gcctctggag
     5521 atcacaaagg agatctacga cctgaacaat cctgagaagg agccaaagaa gtttcagaca
     5581 gcctacgcca agaaaaccgg cgaccagaag ggctacagag aggccctgtg caagtggatc
     5641 gacttcacaa gggattttct gtccaagtat accaagacaa cctctatcga tctgtctagc
     5701 ctgcggccat cctctcagta taaggacctg ggcgagtact atgccgagct gaatcccctg
     5761 ctgtaccaca tcagcttcca gagaatcgcc gagaaggaga tcatggatgc cgtggagaca
     5821 ggcaagctgt acctgttcca gatctataac aaggactttg ccaagggcca ccacggcaag
     5881 cctaatctgc acacactgta ttggaccggc ctgttttctc cagagaacct ggccaagaca
     5941 agcatcaagc tgaatggcca ggccgagctg ttctaccgcc ctaagtccag gatgaagagg
     6001 atggcacacc ggctgggaga gaagatgctg aacaagaagc tgaaggatca gaaaacccca
     6061 atccccgaca ccctgtacca ggagctgtac gactatgtga atcacagact gtcccacgac
     6121 ctgtctgatg aggccagggc cctgctgccc aacgtgatca ccaaggaggt gtctcacgag
     6181 atcatcaagg ataggcgctt taccagcgac aagttctttt tccacgtgcc tatcacactg
     6241 aactatcagg ccgccaattc cccatctaag ttcaaccaga gggtgaatgc ctacctgaag
     6301 gagcaccccg agacacctat catcggcatc gatcggggcg agagaaacct gatctatatc
     6361 acagtgatcg actccaccgg caagatcctg gagcagcgga gcctgaacac catccagcag
     6421 tttgattacc agaagaagct ggacaacagg gagaaggaga gggtggcagc aaggcaggcc
     6481 tggtctgtgg tgggcacaat caaggatctg aagcagggct atctgagcca ggtcatccac
     6541 gagatcgtgg acctgatgat ccactaccag gccgtggtgg tgctggagaa cctgaatttc
     6601 ggctttaaga gcaagaggac cggcatcgcc gagaaggccg tgtaccagca gttcgagaag
     6661 atgctgatcg ataagctgaa ttgcctggtg ctgaaggact atccagcaga gaaagtggga
     6721 ggcgtgctga acccatacca gctgacagac cagttcacct cctttgccaa gatgggcacc
     6781 cagtctggct tcctgtttta cgtgcctgcc ccatatacat ctaagatcga tcccctgacc
     6841 ggcttcgtgg accccttcgt gtggaaaacc atcaagaatc acgagagccg caagcacttc
     6901 ctggagggct tcgactttct gcactacgac gtgaaaaccg gcgacttcat cctgcacttt
     6961 aagatgaaca gaaatctgtc cttccagagg ggcctgcccg gctttatgcc tgcatgggat
     7021 atcgtgttcg agaagaacga gacacagttt gacgccaagg gcaccccttt catcgccggc
     7081 aagagaatcg tgccagtgat cgagaatcac agattcaccg gcagataccg ggacctgtat
     7141 cctgccaacg agctgatcgc cctgctggag gagaagggca tcgtgttcag ggatggctcc
     7201 aacatcctgc caaagctgct ggagaatgac gattctcacg ccatcgacac catggtggcc
     7261 ctgatccgca gcgtgctgca gatgcggaac tccaatgccg ccacaggcga ggactatatc
     7321 aacagccccg tgcgcgatct gaatggcgtg tgcttcgact cccggtttca gaacccagag
     7381 tggcccatgg acgccgatgc caatggcgcc taccacatcg ccctgaaggg ccagctgctg
     7441 ctgaatcacc tgaaggagag caaggatctg aagctgcaga acggcatctc caatcaggac
     7501 tggctggcct acatccagga gctgcgcaac gggtcaccta agaaaaaacg aaaagttgag
     7561 gatcctaaaa agaaacgaaa agttgatacc ggttctaaag gtgaagaact gttcaccggt
     7621 gttgttccga tcctggttga actggatggt gatgttaacg gccacaaatt ctctgttcgt
     7681 ggtgaaggtg aaggtgatgc aaccaacggt aaactgaccc tgaaattcat ctgcactacc
     7741 ggtaaactgc cggttccatg gccgactctg gtgactaccc tgacctatgg tgttcagtgt
     7801 ttttctcgtt acccggatca catgaagcag catgatttct tcaaatctgc aatgccggaa
     7861 ggttatgtac aggagcgcac catttctttc aaagacgatg gcacctacaa aacccgtgca
     7921 gaggttaaat ttgaaggtga tactctggtg aaccgtattg aactgaaagg cattgatttc
     7981 aaagaggacg gcaacatcct gggccacaaa ctggaatata acttcaactc ccataacgtt
     8041 tacatcaccg cagacaaaca gaagaacggt atcaaagcta acttcaaaat tcgccataac
     8101 gttgaagacg gtagcgtaca gctggcggac cactaccagc agaacactcc gatcggtgat
     8161 ggtccggttc tgctgccgga taaccactac ctgtccaccc agtctgttct gtccaaagac
     8221 ccgaacgaaa agcgcgacca catggtgctg ctggagttcg ttactgcagc aggtatcacg
     8281 cacggcatgg atgagctcta caaataagcg gccgcactcg aggcccgaaa ggaagctgag
     8341 ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc
     8401 ttgaggggtt ttttgctgaa aggaggaact atatccggat atcccgcaag aggcccggca
     8461 gtaccggcat aaccaagcct atgcctacag catccagggt gacggtgccg aggatgacga
     8521 tgagcgcatt gttagatttc atacacggtg cctgactgcg ttagcaattt aactgtgata
     8581 aactaccgca ttaaagctta tcgatgataa gctgtcaaac atgagaattc ttgaagacga
     8641 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     8701 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     8761 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     8821 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     8881 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     8941 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     9001 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     9061 ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat
     9121 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     9181 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     9241 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     9301 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     9361 cgtgacacca cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa
     9421 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     9481 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     9541 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     9601 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     9661 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     9721 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     9781 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     9841 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     9901 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     9961 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
    10021 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
    10081 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
    10141 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
    10201 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
    10261 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
    10321 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
    10381 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
    10441 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
    10501 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
    10561 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
    10621 agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
    10681 tcacaccgca atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
    10741 atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc
    10801 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
    10861 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca
    10921 gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc
    10981 cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc
    11041 catgttaagg gcggttt
//