LOCUS       sequence-71838-e        8813 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Mammalian S. thermophilus #1 Cas9 expression, human optimized.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8813)
  AUTHORS   Esvelt KM, Mali P, Braff JL, Moosburner M, Yaung SJ, Church GM
  TITLE     Orthogonal Cas9 proteins for RNA-guided gene regulation and editing.
  JOURNAL   Nat Methods. 2013 Sep 29. doi: 10.1038/nmeth.2681.
  PUBMED    24076762
REFERENCE   2  (bases 1 to 8813)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Methods. 2013 Sep 29. doi: 10.1038/nmeth.2681."
COMMENT     Sequence Label: M-ST1cas
FEATURES             Location/Qualifiers
     source          1..8813
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        50..429
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        430..633
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     584..604
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     630..654
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             748..4110
                     /codon_start=1
                     /product="Cas9 endonuclease from the Streptococcus 
                     thermophilus Type II CRISPR/Cas system (Esvelt et al., 
                     2013)"
                     /label=St1Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MSDLVLGLDIGIGSVGVGILNKVTGEIIHKNSRIFPAAQAENNLV
                     RRTNRQGRRLARRKKHRRVRLNRLFEESGLITDFTKISINLNPYQLRVKGLTDELSNEE
                     LFIALKNMVKHRGISYLDDASDDGNSSVGDYAQIVKENSKQLETKTPGQIQLERYQTYG
                     QLRGDFTVEKDGKKHRLINVFPTSAYRSEALRILQTQQEFNPQITDEFINRYLEILTGK
                     RKYYHGPGNEKSRTDYGRYRTSGETLDNIFGILIGKCTFYPDEFRAAKASYTAQEFNLL
                     NDLNNLTVPTETKKLSKEQKNQIINYVKNEKAMGPAKLFKYIAKLLSCDVADIKGYRID
                     KSGKAEIHTFEAYRKMKTLETLDIEQMDRETLDKLAYVLTLNTEREGIQEALEHEFADG
                     SFSQKQVDELVQFRKANSSIFGKGWHNFSVKLMMELIPELYETSEEQMTILTRLGKQKT
                     TSSSNKTKYIDEKLLTEEIYNPVVAKSVRQAIKIVNAAIKEYGDFDNIVIEMARETNED
                     DEKKAIQKIQKANKDEKDAAMLKAANQYNGKAELPHSVFHGHKQLATKIRLWHQQGERC
                     LYTGKTISIHDLINNSNQFEVDHILPLSITFDDSLANKVLVYATANQEKGQRTPYQALD
                     SMDDAWSFRELKAFVRESKTLSNKKKEYLLTEEDISKFDVRKKFIERNLVDTLYASRVV
                     LNALQEHFRAHKIDTKVSVVRGQFTSQLRRHWGIEKTRDTYHHHAVDALIIAASSQLNL
                     WKKQKNTLVSYSEDQLLDIETGELISDDEYKESVFKAPYQHFVDTLKSKEFEDSILFSY
                     QVDSKFNRKISDATIYATRQAKVGKDKADETYVLGKIKDIYTQDGYDAFMKIYKKDKSK
                     FLMYRHDPQTFEKVIEPILENYPNKQINDKGKEVPCNPFLKYKEEHGYIRKYSKKGNGP
                     EIKSLKYYDSKLGNHIDITPKDSNNKVVLQSVSPWRADVYFNKTTGKYEILGLKYADLQ
                     FDKGTGTYKISQEKYNDIKKKEGVDSDSEFKFTLYKNDLLLVKDTETKEQQLFRFLSRT
                     MPKQKHYVELKPYDKQKFEGGEALIKVLGNVANSGQCKKGLGKSNISIYKVRTDVLGNQ
                     HIIKNEGDKPKLDF"
     CDS             4123..4143
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4199..4218)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4243..4291
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4493..4921
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4580..4599)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4790..4811
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4930..4950)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4935..5264
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5115..5250
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5177..5196
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5331..6125
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5385..5404)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5995..6014
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6301..6422
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6338..6357)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6392..6411
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6471..6487)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6471..6487)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6484..6506)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6495..6511
                     /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(join(6519..6525,6526..6543,6544..6549))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6564..6585
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6702..6719)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6873..7461)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6953..6972)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7632..8423,8424..8492))
                     /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     8255..8274
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8493..8597)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8672..8691)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg
      661 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg
      721 actctagagg atcgaaccct tgccaccatg agcgacctgg tgctgggcct ggacatcggc
      781 atcggcagcg tgggcgtggg catcctgaac aaggtgaccg gcgagatcat ccacaagaac
      841 agtcgcatct tccctgctgc tcaggctgag aacaacctgg tgcgccgcac caaccgccag
      901 ggtcgccggc ttgctcgccg caagaagcac cggcgcgtgc gcctgaaccg cctgttcgag
      961 gagagcggcc tgatcaccga cttcaccaag atcagcatca acctgaaccc ctaccagctg
     1021 cgcgtgaagg gcctgaccga cgagctgagc aacgaggagc tgttcatcgc cctgaagaac
     1081 atggtgaagc accgcggcat cagctacctg gacgacgcca gcgacgacgg caacagcagc
     1141 gtgggcgact acgcccagat cgtgaaggag aacagcaagc agctggagac caagaccccc
     1201 ggccagatcc agctggagcg ctaccagacc tacggccagc tgcgcggcga cttcaccgtg
     1261 gagaaggacg gcaagaagca ccgcctgatc aacgtgttcc ccaccagcgc ctaccgcagc
     1321 gaggccctgc gcatcctgca gacccagcag gagttcaacc cccagatcac cgacgagttc
     1381 atcaaccgct acctggagat cctgaccggc aagcgcaagt actaccacgg ccccggcaac
     1441 gagaagagcc gcaccgacta cggccgctac cgcaccagcg gcgagaccct ggacaacatc
     1501 ttcggcatcc tgatcggcaa gtgcaccttc taccccgacg agttccgcgc cgccaaggcc
     1561 agctacaccg cccaggagtt caacctgctg aacgacctga acaacctgac cgtgcccacc
     1621 gagaccaaga agctgagcaa ggagcagaag aaccagatca tcaactacgt gaagaacgag
     1681 aaggccatgg gccccgccaa gctgttcaag tacatcgcca agctgctgag ctgcgacgtg
     1741 gccgacatca agggctaccg catcgacaag agcggcaagg ccgagatcca caccttcgag
     1801 gcctaccgca agatgaagac cctggagacc ctggacatcg agcagatgga ccgcgagacc
     1861 ctggacaagc tggcctacgt gctgaccctg aacaccgagc gcgagggcat ccaggaggcc
     1921 ctggagcacg agttcgccga cggcagcttc agccagaagc aggtggacga gctggtgcag
     1981 ttccgcaagg ccaacagcag catcttcggc aagggctggc acaacttcag cgtgaagctg
     2041 atgatggagc tgatccccga gctgtacgag accagcgagg agcagatgac catcctgacc
     2101 cgcctgggca agcagaagac caccagcagc agcaacaaga ccaagtacat cgacgagaag
     2161 ctgctgaccg aggagatcta caaccccgtg gtggccaaga gcgtgcgcca ggccatcaag
     2221 atcgtgaacg ccgccatcaa ggagtacggc gacttcgaca acatcgtgat cgagatggcc
     2281 cgcgagacca acgaggacga cgagaagaag gccatccaga agatccagaa ggccaacaag
     2341 gacgagaagg acgccgccat gctgaaggcc gccaaccagt acaacggcaa ggccgagctg
     2401 ccccacagcg tgttccacgg ccacaagcag ctggccacca agatccgcct gtggcaccag
     2461 cagggcgagc gctgcctgta caccggcaag accatcagca tccacgacct gatcaacaac
     2521 agcaaccagt tcgaggtgga ccacatcctg cccctgagca tcaccttcga cgacagcctg
     2581 gccaacaagg tgctggtgta cgccaccgcc aaccaggaga agggccagcg caccccctac
     2641 caggccctgg acagcatgga cgacgcctgg agcttccgcg agctgaaggc cttcgtgcgc
     2701 gagagcaaga ccctgagcaa caagaagaag gagtacctgc tgaccgagga ggacatcagc
     2761 aagttcgacg tgcgcaagaa gttcatcgag cgcaacctgg tggacaccct ctacgccagc
     2821 cgcgtggtgc tgaacgccct gcaggagcac ttccgcgccc acaagatcga caccaaggtg
     2881 agcgtggtgc gcggccagtt caccagccag ctgcgccgcc actggggcat cgagaagacc
     2941 cgcgacacct accaccacca cgccgtggac gccctgatca ttgcggcttc tagccagctg
     3001 aacctgtgga agaagcagaa gaacaccctg gtgagctaca gcgaggacca gctgctggac
     3061 atcgagaccg gcgagctgat cagcgacgac gagtacaagg agagcgtgtt caaggccccc
     3121 taccagcact tcgtggacac cctgaagagc aaggagttcg aggacagcat cctgttcagc
     3181 taccaggtgg acagcaagtt caaccgcaag atcagcgacg ccaccatcta cgccacccgc
     3241 caggccaagg tgggcaagga caaggccgac gagacctacg tgctgggcaa gatcaaggac
     3301 atctacaccc aggacggcta cgacgccttc atgaagatct acaagaagga caagagcaag
     3361 ttcctgatgt accgccacga cccccagacc ttcgagaagg tgatcgagcc catcctggag
     3421 aactacccca acaagcagat caacgataaa ggcaaggagg tgccctgcaa ccccttcctg
     3481 aagtacaagg aggagcacgg ctacatccgc aagtacagca agaagggcaa cggccccgag
     3541 atcaagagcc tgaagtacta cgacagcaag ctgggcaacc acatcgacat cacccccaag
     3601 gacagcaaca acaaggtggt gctgcagagc gtgagcccct ggcgcgccga cgtgtacttc
     3661 aacaagacca ccggcaagta cgagatcctg ggcctgaagt acgccgacct gcagtttgat
     3721 aagggcaccg gcacctacaa gatcagccag gagaagtaca acgacatcaa gaagaaggag
     3781 ggcgtggaca gcgacagcga gttcaagttc accctgtaca agaacgacct tctgctggtg
     3841 aaggacaccg agaccaagga gcaacagctg ttccgcttcc tgagccgcac catgcccaag
     3901 cagaagcact acgtggagct gaagccctac gacaagcaga agttcgaggg cggcgaggcc
     3961 ctgatcaagg tgctgggcaa cgtggccaac agcggccagt gcaagaaggg cctgggcaag
     4021 agcaacatca gcatctacaa ggtgcgcacc gacgtgctgg gcaaccagca catcatcaag
     4081 aacgagggcg acaagcccaa gttggacttc agcagggctg accccaagaa gaagaggaag
     4141 gtgtgagcag ggttcgatcc ctaccggtta gtaatgagtt taaacggggg aggctaactg
     4201 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     4261 ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg
     4321 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt
     4381 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg
     4441 gcagaccctg ccatagcaga tctgcgcagc tggggctcta gggggtatcc ccacgcgccc
     4501 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     4561 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     4621 ggctttcccc gtcaagctct aaatcggggc atccctttag ggttccgatt tagtgcttta
     4681 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     4741 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     4801 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     4861 ttggggattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     4921 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     4981 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     5041 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     5101 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     5161 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     5221 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     5281 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     5341 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     5401 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     5461 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     5521 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     5581 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     5641 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     5701 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     5761 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     5821 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     5881 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     5941 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     6001 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     6061 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     6121 tctgagcggg actctggggt tcgcgaaatg accgaccaag cgacgcccaa cctgccatca
     6181 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     6241 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     6301 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     6361 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     6421 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     6481 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     6541 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     6601 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     6661 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     6721 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     6781 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     6841 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     6901 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     6961 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     7021 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt
     7081 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     7141 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     7201 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     7261 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     7321 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     7381 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     7441 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     7501 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     7561 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     7621 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     7681 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     7741 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     7801 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     7861 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     7921 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     7981 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     8041 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     8101 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     8161 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     8221 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     8281 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     8341 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     8401 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     8461 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     8521 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     8581 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
     8641 gatctcccga tcccctatgg tcgactctca gtacaatctg ctctgatgcc gcatagttaa
     8701 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
     8761 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agg
//
