LOCUS       sequence-217478-       10440 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Knockout eGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10440)
  AUTHORS   Sinha H, Quach ABV, Vo PQN, Shih SCC
  TITLE     An automated microfluidic gene-editing platform for deciphering 
            cancer genes.
  JOURNAL   Lab Chip. 2018 Jul 24;18(15):2300-2312. doi: 10.1039/c8lc00470f.
  PUBMED    29989627
REFERENCE   2  (bases 1 to 10440)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10"; journalName: 
            "Lab Chip"; date: "2018-07-24- 24"; volume: "18"; issue: "15"; 
            pages: "2300-2312"
COMMENT     Sequence Label: pCRISPR_eGFP_497
FEATURES             Location/Qualifiers
     source          1..10440
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     121..138
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    255..276
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(291..296,297..314,315..321)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    329..345
                     /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)."
     primer_bind     353..369
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        389..407
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     389..406
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        419..437
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        568..808
                     /label=U6 promoter
                     /note="Truncated RNA polymerase III promoter for human U6 
                     snRNA used in pSIREN vectors such as 
                     pSIREN-RetroQ-ZsGreen1. Expected transcription start site 
                     is 8bp downstream of this feature."
     primer_bind     568..588
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     739..758
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        837..912
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        1000..1379
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1380..1583
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1534..1554
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        1628..1646
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1660..1669
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1666..5769
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             1666..5769
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             5785..5805
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5806..5832
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             5806..5832
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(5849..5869)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5854..6183
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6034..6169
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6096..6115
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     regulatory      6241..6250
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6247..6957
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(6392..6410)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     6660..6679
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    join(6972..7546,7547..7549,7550..7558)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7139..7156)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             7581..8384
                     /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(7644..7663)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8254..8273
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    8516..8637
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8553..8572)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8607..8626
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8686..8702)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    8710..8726
                     /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)."
     CDS             join(8788..8856,8857..9648)
                     /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     complement(9006..9025)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9819..10407
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10308..10327
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
       61 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
      121 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
      181 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
      241 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
      301 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
      361 agctatgaca tgattacgaa ttgcaacgat ttaggtgaca ctatagaaga gaaggaatta
      421 atacgactca ctatagggag agagagagaa ttaccctcac taaagggagg agaagcatga
      481 attccccagt ggaaagacgc gcaggcaaaa cgcaccacgt gacggagcgt gaccgcgcgc
      541 cgagcgcgcg ccaaggtcgg gcaggaagag ggcctatttc ccatgattcc ttcatatttg
      601 catatacgat acaaggctgt tagagagata attagaatta atttgactgt aaacacaaag
      661 atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt tgcagtttta
      721 aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta tttcgatttc
      781 ttgggtttat atatcttgtg gaaaggacga ggatctcaag atccgccaca acatcggttt
      841 tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca
      901 ccgagtcggt gcttttttct agacacaatt gcatgaagaa tctgcttagg gttaggcgtt
      961 ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt
     1021 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     1081 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     1141 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     1201 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     1261 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     1321 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
     1381 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     1441 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     1501 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     1561 gggaggtcta tataagcaga gctctctggc taactagaga acccactgct tactggctta
     1621 tcgaaattaa tacgactcac tatagggaga cccaagcttg ccaccatgga caagaagtac
     1681 agcatcggcc tggacatcgg taccaacagc gtgggctggg ccgtgatcac cgacgagtac
     1741 aaggtgccca gcaagaagtt caaggtgctg ggcaacaccg accgccacag catcaagaag
     1801 aacctgatcg gcgccctgct gttcgacagc ggcgagaccg ccgaggccac ccgcctgaag
     1861 cgcaccgccc gccgccgcta cacccgccgc aagaaccgca tctgctacct gcaggagatc
     1921 ttcagcaacg agatggccaa ggtggacgac agcttcttcc accgcctgga ggagagcttc
     1981 ctggtggagg aggacaagaa gcacgagcgc caccccatct tcggcaacat cgtggacgag
     2041 gtggcctacc acgagaagta ccccaccatc taccacctgc gcaagaagct ggtggacagc
     2101 accgacaagg ccgacctgcg cctgatctac ctggccctgg cccacatgat caagttccgc
     2161 ggccacttcc tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga caagctgttc
     2221 atccagctgg tgcagaccta caaccagctg ttcgaggaga accccatcaa cgccagcggc
     2281 gtggacgcca aggccatcct gagcgcccgc ctgagcaaga gccgccgcct ggagaacctg
     2341 atcgcccagc tgcccggcga gaagaagaac ggcctgttcg gcaacctgat cgccctgagc
     2401 ctgggcctga cccccaactt caagagcaac ttcgacctgg ccgaggacgc caagctgcag
     2461 ctgagcaagg acacctacga cgacgacctg gacaacctgc tggcccagat cggcgaccag
     2521 tacgccgacc tgttcctggc cgccaagaac ctgagcgacg ccatcctgct gagcgacatc
     2581 ctgcgcgtga acaccgagat caccaaggcc cccctgagcg ccagcatgat caagcgctac
     2641 gacgagcacc accaggacct gaccctgctg aaggccctgg tgcgccagca gctgcccgag
     2701 aagtacaagg agatcttctt cgaccagagc aagaacggct acgccggcta catcgacggc
     2761 ggcgccagcc aggaggagtt ctacaagttc atcaagccca tcctggagaa gatggacggc
     2821 accgaggagc tgctggtgaa gctgaaccgc gaggacctgc tgcgcaagca gcgcaccttc
     2881 gacaacggca gcatccccca ccagatccac ctgggcgagc tgcacgccat cctgcgccgc
     2941 caggaggact tctacccctt cctgaaggac aaccgcgaga agatcgagaa gatcctgacc
     3001 ttccgcatcc cctactacgt gggccccctg gcccgcggca acagccgctt cgcctggatg
     3061 acccgcaaga gcgaggagac catcaccccc tggaacttcg aggaggtggt ggacaagggc
     3121 gccagcgccc agagcttcat cgagcgcatg accaacttcg acaagaacct gcccaacgag
     3181 aaggtgctgc ccaagcacag cctgctgtac gagtacttca ccgtgtacaa cgagctgacc
     3241 aaggtgaagt acgtgaccga gggcatgcgc aagcccgcct tcctgagcgg cgagcagaag
     3301 aaggccatcg tggacctgct gttcaagacc aaccgcaagg tgaccgtgaa gcagctgaag
     3361 gaggactact tcaagaagat cgagtgcttc gacagcgtgg agatcagcgg cgtggaggac
     3421 cgcttcaacg ccagcctggg cacctaccac gacctgctga agatcatcaa ggacaaggac
     3481 ttcctggaca acgaggagaa cgaggacatc ctggaggaca tcgtgctgac cctgaccctg
     3541 ttcgaggacc gcgagatgat cgaggagcgc ctgaagacct acgcccacct gttcgacgac
     3601 aaggtgatga agcagctgaa gcgccgccgc tacaccggct ggggccgcct gagccgcaag
     3661 cttatcaacg gcatccgcga caagcagagc ggcaagacca tcctggactt cctgaagagc
     3721 gacggcttcg ccaaccgcaa cttcatgcag ctgatccacg acgacagcct gaccttcaag
     3781 gaggacatcc agaaggccca ggtgagcggc cagggcgaca gcctgcacga gcacatcgcc
     3841 aacctggccg gcagccccgc catcaagaag ggcatcctgc agaccgtgaa ggtggtggac
     3901 gagctggtga aggtgatggg ccgccacaag cccgagaaca tcgtgatcga gatggcccgc
     3961 gagaaccaga ccacccagaa gggccagaag aacagccgcg agcgcatgaa gcgcatcgag
     4021 gagggcatca aggagctggg cagccagatc ctgaaggagc accccgtgga gaacacccag
     4081 ctgcagaacg agaagctgta cctgtactac ctgcagaacg gccgcgacat gtacgtggac
     4141 caggagctgg acatcaaccg cctgagcgac tacgacgtgg accacatcgt gccccagagc
     4201 ttcctgaagg acgacagcat cgacaacaag gtgctgaccc gcagcgacaa gaaccgcggc
     4261 aagagcgaca acgtgcccag cgaggaggtg gtgaagaaga tgaagaacta ctggcgccag
     4321 ctgctgaacg ccaagctgat cacccagcgc aagttcgaca acctgaccaa ggccgagcgc
     4381 ggcggcctga gcgagctgga caaggccggc ttcatcaagc gccagctggt ggagacccgc
     4441 cagatcacca agcacgtggc ccagatcctg gacagccgca tgaacaccaa gtacgacgag
     4501 aacgacaagc tgatccgcga ggtgaaggtg atcaccctga agagcaagct ggtgagcgac
     4561 ttccgcaagg acttccagtt ctacaaggtg cgcgagatca acaactacca ccacgcccac
     4621 gacgcctacc tgaacgccgt ggtgggcacc gccctgatca agaagtaccc caagctggag
     4681 agcgagttcg tgtacggcga ctacaaggtg tacgacgtgc gcaagatgat cgccaagagc
     4741 gagcaggaga tcggcaaggc caccgccaag tacttcttct acagcaacat catgaacttc
     4801 ttcaagaccg agatcaccct ggccaacggc gagatccgca agcgccccct gatcgagacc
     4861 aacggcgaga ccggcgagat cgtgtgggac aagggccgcg acttcgccac cgtgcgcaag
     4921 gtgctgagca tgccccaggt gaacatcgtg aagaagaccg aggtgcagac cggcggcttc
     4981 agcaaggaga gcatcctgcc caagcgcaac agcgacaagc tgatcgcccg caagaaggac
     5041 tgggacccca agaagtacgg cggcttcgac agccccaccg tggcctacag cgtgctggtg
     5101 gtggccaagg tggagaaggg caagagcaag aagctgaaga gcgtgaagga gctgctgggc
     5161 atcaccatca tggagcgcag cagcttcgag aagaacccca tcgacttcct ggaggccaag
     5221 ggctacaagg aggtgaagaa ggacctgatc atcaagctgc ccaagtacag cctgttcgag
     5281 ctggagaacg gccgcaagcg catgctggcc agcgccggcg agctgcagaa gggcaacgag
     5341 ctggccctgc ccagcaagta cgtgaacttc ctgtacctgg ccagccacta cgagaagctg
     5401 aagggcagcc ccgaggacaa cgagcagaag cagctgttcg tggagcagca caagcactac
     5461 ctggacgaga tcatcgagca gatcagcgag ttcagcaagc gcgtgatcct ggccgacgcc
     5521 aacctggaca aggtgctgag cgcctacaac aagcaccgcg acaagcccat ccgcgagcag
     5581 gccgagaaca tcatccacct gttcaccctg accaacctgg gcgcccccgc cgccttcaag
     5641 tacttcgaca ccaccatcga ccgcaagcgc tacaccagca ccaaggaggt gctggacgcc
     5701 accctgatcc accagagcat caccggtctg tacgagaccc gcatcgacct gagccagctg
     5761 ggcggcgacg gcggctccgg acctccaaag aaaaagagaa aagtataccc ctacgacgtg
     5821 cccgactacg cctaataact cgagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     5881 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     5941 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     6001 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     6061 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     6121 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     6181 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcggcgcgc cataccggtc
     6241 gccaccatgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc
     6301 ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
     6361 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc
     6421 ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg
     6481 aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg
     6541 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg
     6601 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc
     6661 cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag
     6721 gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac
     6781 gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac
     6841 aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag
     6901 tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtagcgg
     6961 ccgcgatccc gcccctctcc ctcccccccc cctaacgtta ctggccgaag ccgcttggaa
     7021 taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat
     7081 gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct
     7141 ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct
     7201 tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc
     7261 gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa
     7321 ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct cacctcaagc
     7381 gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg
     7441 gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc
     7501 ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataagcttg ccacaacccg
     7561 cgataattcc tgcagccaat atgggatcgg ccattgaaca agatggattg cacgcaggtt
     7621 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     7681 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     7741 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     7801 ccacgacggg cgttccttgc gcagcagtgc tcgacgttgt cactgaagcg ggaagggact
     7861 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     7921 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     7981 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     8041 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     8101 tcgccaggct caaggcgcgt atgcccgacg gcgatgatct cgtcgtgact catggcgatg
     8161 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     8221 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     8281 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     8341 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgaacgcgg tgctacgaga
     8401 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     8461 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     8521 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     8581 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     8641 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     8701 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaaggt
     8761 aagcctgaat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     8821 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     8881 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     8941 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     9001 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     9061 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     9121 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     9181 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     9241 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     9301 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     9361 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     9421 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     9481 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     9541 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     9601 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     9661 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     9721 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     9781 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     9841 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttt tttgccggat
     9901 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     9961 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
    10021 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
    10081 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
    10141 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
    10201 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
    10261 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
    10321 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
    10381 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
//
