LOCUS       sequence-376613-       12707 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Cas9-P2A-Eco1RT in human cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12707)
  AUTHORS   Lopez SC, Crawford KD, Lear SK, Bhattarai-Kline S, Shipman SL
  TITLE     Precise genome editing across kingdoms of life using retron-derived 
            DNA.
  JOURNAL   Nat Chem Biol. 2022 Feb;18(2):199-206. doi: 
            10.1038/s41589-021-00927-y. Epub 2021 Dec 23.
  PUBMED    34949838
REFERENCE   2  (bases 1 to 12707)
  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: 
            "10.1038/s41589-021-00927-y"; journalName: "Nat Chem Biol"; date: 
            "2022-02"; volume: "18"; issue: "2"; pages: "199-206"
COMMENT     Sequence Label: pSCL.139
FEATURES             Location/Qualifiers
     source          1..12707
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..105)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(131..586)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(268..289)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     480..499
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     713..735
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     727..744
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     728..744
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   874..936
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        1385..1763
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    1393..1411
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1429..1447
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1465..1483
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1501..1519
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1537..1555
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1573..1591
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1609..1627
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1672..1696
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1798..1818
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1819..5919
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             1819..5919
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             1819..5919
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5920..5976
                     /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             6937..6957
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(6998..7017)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7042..7090
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        7321..7532
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             7545..7813
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             7847..8590
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     CDS             8591..8644
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             8645..9244
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(8645..8664)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     9141..9161
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    9277..9398
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9314..9333)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9368..9387
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   10161..10195
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(10686..10702)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10686..10702)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10699..10721)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10710..10726
                     /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(10734..10740,10741..10758,10759..10764))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10779..10800
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10917..10934)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11088..11676)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11168..11187)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11847..12638,12639..12707))
                     /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     12470..12489
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
      181 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      241 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
      301 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
      361 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
      421 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
      481 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
      541 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
      601 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
      661 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
      721 cgacgttgta aaacgacggc cagtgagcgc gcctcgttca ttcacgtttt tgaacccgtg
      781 gaggacgggc agactcgcgg tgcaaatgtg ttttacagcg tgatggagca gatgaagatg
      841 ctcgacacgc tgcagaacac gcagctagat taaccctaga aagataatca tattgtgacg
      901 tacgttaaag ataatcatgt gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag
      961 gtttatttat taatttgaat agatattaag ttttattata tttacactta catactaata
     1021 ataaattcaa caaacaattt atttatgttt atttatttat taaaaaaaac aaaaactcaa
     1081 aatttcttct ataaagtaac aaaactttta tgagggacag ccccccccca aagcccccag
     1141 ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg agccgcccgg ggctccgctc
     1201 cggtccggcg ctccccccgc atccccgagc cggcagcgtg cggggacagc ccgggcacgg
     1261 ggaaggtggc acgggatcgc tttcctctga acgcttctcg ctgctctttg agcctgcaga
     1321 cacctggggg gatacgggga aaaggcctcc acggccacta gtctttcgtc ttcaagaatt
     1381 cctcgagttt actccctatc agtgatagag aacgtatgaa gagtttactc cctatcagtg
     1441 atagagaacg tatgcagact ttactcccta tcagtgatag agaacgtata aggagtttac
     1501 tccctatcag tgatagagaa cgtatgacca gtttactccc tatcagtgat agagaacgta
     1561 tctacagttt actccctatc agtgatagag aacgtatatc cagtttactc cctatcagtg
     1621 atagagaacg tataagcttt aggcgtgtac ggtgggcgcc tataaaagca gagctcgttt
     1681 agtgaaccgt cagatcgcct ggagcaattc cacaacactt ttgtcttata ccaactttcc
     1741 gtaccacttc ctaccctcgt aaaggtctag agctaggcca ccatgagcag ggctgacccc
     1801 aagaagaaga ggaaggtgga caagaagtac tccattgggc tcgatatcgg cacaaacagc
     1861 gtcggttggg ccgtcattac ggacgagtac aaggtgccga gcaaaaaatt caaagttctg
     1921 ggcaataccg atcgccacag cataaagaag aacctcattg gcgccctcct gttcgactcc
     1981 ggggagacgg ccgaagccac gcggctcaaa agaacagcac ggcgcagata tacccgcaga
     2041 aagaatcgga tctgctacct gcaggagatc tttagtaatg agatggctaa ggtggatgac
     2101 tctttcttcc ataggctgga ggagtccttt ttggtggagg aggataaaaa gcacgagcgc
     2161 cacccaatct ttggcaatat cgtggacgag gtggcgtacc atgaaaagta cccaaccata
     2221 tatcatctga ggaagaagct tgtagacagt actgataagg ctgacttgcg gttgatctat
     2281 ctcgcgctgg cgcatatgat caaatttcgg ggacacttcc tcatcgaggg ggacctgaac
     2341 ccagacaaca gcgatgtcga caaactcttt atccaactgg ttcagactta caatcagctt
     2401 ttcgaagaga acccgatcaa cgcatccgga gttgacgcca aagcaatcct gagcgctagg
     2461 ctgtccaaat cccggcggct cgaaaacctc atcgcacagc tccctgggga gaagaagaac
     2521 ggcctgtttg gtaatcttat cgccctgtca ctcgggctga cccccaactt taaatctaac
     2581 ttcgacctgg ccgaagatgc caagcttcaa ctgagcaaag acacctacga tgatgatctc
     2641 gacaatctgc tggcccagat cggcgaccag tacgcagacc tttttttggc ggcaaagaac
     2701 ctgtcagacg ccattctgct gagtgatatt ctgcgagtga acacggagat caccaaagct
     2761 ccgctgagcg ctagtatgat caagcgctat gatgagcacc accaagactt gactttgctg
     2821 aaggcccttg tcagacagca actgcctgag aagtacaagg aaattttctt cgatcagtct
     2881 aaaaatggct acgccggata cattgacggc ggagcaagcc aggaggaatt ttacaaattt
     2941 attaagccca tcttggaaaa aatggacggc accgaggagc tgctggtaaa gcttaacaga
     3001 gaagatctgt tgcgcaaaca gcgcactttc gacaatggaa gcatccccca ccagattcac
     3061 ctgggcgaac tgcacgctat cctcaggcgg caagaggatt tctacccctt tttgaaagat
     3121 aacagggaaa agattgagaa aatcctcaca tttcggatac cctactatgt aggccccctc
     3181 gcccggggaa attccagatt cgcgtggatg actcgcaaat cagaagagac catcactccc
     3241 tggaacttcg aggaagtcgt ggataagggg gcctctgccc agtccttcat cgaaaggatg
     3301 actaactttg ataaaaatct gcctaacgaa aaggtgcttc ctaaacactc tctgctgtac
     3361 gagtacttca cagtttataa cgagctcacc aaggtcaaat acgtcacaga agggatgaga
     3421 aagccagcat tcctgtctgg agagcagaag aaagctatcg tggacctcct cttcaagacg
     3481 aaccggaaag ttaccgtgaa acagctcaaa gaagactatt tcaaaaagat tgaatgtttc
     3541 gactctgttg aaatcagcgg agtggaggat cgcttcaacg catccctggg aacgtatcac
     3601 gatctcctga aaatcattaa agacaaggac ttcctggaca atgaggagaa cgaggacatt
     3661 cttgaggaca ttgtcctcac ccttacgttg tttgaagata gggagatgat tgaagaacgc
     3721 ttgaaaactt acgctcatct cttcgacgac aaagtcatga aacagctcaa gaggcgccga
     3781 tatacaggat gggggcggct gtcaagaaaa ctgatcaatg ggatccgaga caagcagagt
     3841 ggaaagacaa tcctggattt tcttaagtcc gatggatttg ccaaccggaa cttcatgcag
     3901 ttgatccatg atgactctct cacctttaag gaggacatcc agaaagcaca agtttctggc
     3961 cagggggaca gtcttcacga gcacatcgct aatcttgcag gtagcccagc tatcaaaaag
     4021 ggaatactgc agaccgttaa ggtcgtggat gaactcgtca aagtaatggg aaggcataag
     4081 cccgagaata tcgttatcga gatggcccga gagaaccaaa ctacccagaa gggacagaag
     4141 aacagtaggg aaaggatgaa gaggattgaa gagggtataa aagaactggg gtcccaaatc
     4201 cttaaggaac acccagttga aaacacccag cttcagaatg agaagctcta cctgtactac
     4261 ctgcagaacg gcagggacat gtacgtggat caggaactgg acatcaatcg gctctccgac
     4321 tacgacgtgg atcatatcgt gccccagtct tttctcaaag atgattctat tgataataaa
     4381 gtgttgacaa gatccgataa aaatagaggg aagagtgata acgtcccctc agaagaagtt
     4441 gtcaagaaaa tgaaaaatta ttggcggcag ctgctgaacg ccaaactgat cacacaacgg
     4501 aagttcgata atctgactaa ggctgaacga ggtggcctgt ctgagttgga taaagccggc
     4561 ttcatcaaaa ggcagcttgt tgagacacgc cagatcacca agcacgtggc ccaaattctc
     4621 gattcacgca tgaacaccaa gtacgatgaa aatgacaaac tgattcgaga ggtgaaagtt
     4681 attactctga agtctaagct ggtctcagat ttcagaaagg actttcagtt ttataaggtg
     4741 agagagatca acaattacca ccatgcgcat gatgcctacc tgaatgcagt ggtaggcact
     4801 gcacttatca aaaaatatcc caagcttgaa tctgaatttg tttacggaga ctataaagtg
     4861 tacgatgtta ggaaaatgat cgcaaagtct gagcaggaaa taggcaaggc caccgctaag
     4921 tacttctttt acagcaatat tatgaatttt ttcaagaccg agattacact ggccaatgga
     4981 gagattcgga agcgaccact tatcgaaaca aacggagaaa caggagaaat cgtgtgggac
     5041 aagggtaggg atttcgcgac agtccggaag gtcctgtcca tgccgcaggt gaacatcgtt
     5101 aaaaagaccg aagtacagac cggaggcttc tccaaggaaa gtatcctccc gaaaaggaac
     5161 agcgacaagc tgatcgcacg caaaaaagat tgggacccca agaaatacgg cggattcgat
     5221 tctcctacag tcgcttacag tgtactggtt gtggccaaag tggagaaagg gaagtctaaa
     5281 aaactcaaaa gcgtcaagga actgctgggc atcacaatca tggagcgatc aagcttcgaa
     5341 aaaaacccca tcgactttct cgaggcgaaa ggatataaag aggtcaaaaa agacctcatc
     5401 attaagcttc ccaagtactc tctctttgag cttgaaaacg gccggaaacg aatgctcgct
     5461 agtgcgggcg agctgcagaa aggtaacgag ctggcactgc cctctaaata cgttaatttc
     5521 ttgtatctgg ccagccacta tgaaaagctc aaagggtctc ccgaagataa tgagcagaag
     5581 cagctgttcg tggaacaaca caaacactac cttgatgaga tcatcgagca aataagcgaa
     5641 ttctccaaaa gagtgatcct cgccgacgct aacctcgata aggtgctttc tgcttacaat
     5701 aagcacaggg ataagcccat cagggagcag gcagaaaaca ttatccactt gtttactctg
     5761 accaacttgg gcgcgcctgc agccttcaag tacttcgaca ccaccataga cagaaagcgg
     5821 tacacctcta caaaggaggt cctggacgcc acactgattc atcagtcaat tacggggctc
     5881 tatgaaacaa gaatcgacct ctctcagctc ggtggagacg caacaaactt ctctctgctg
     5941 aaacaagccg gagatgtcga agagaatcct ggaccgaagt ccgctgaata cttgaacacc
     6001 ttcagattga gaaatttggg tttgccagtc atgaacaact tgcacgatat gtctaaggct
     6061 accagaattt ctgttgaaac cttgagatta ttgatctaca ccgccgattt cagatacaga
     6121 atctacactg ttgaaaagaa gggtccagaa aagagaatga gaactatcta ccaaccatcc
     6181 agagaattga aggctttaca aggttgggtt ttgagaaaca tcttggacaa gttatcctcc
     6241 tccccatttt ctatcggttt tgaaaagcac caatccatct tgaacaatgc cactccacat
     6301 attggtgcca acttcatttt gaacatcgac ttggaagatt tcttcccatc tttgactgct
     6361 aacaaggttt tcggtgtttt ccattctttg ggttacaaca gattgatctc ctctgttttg
     6421 accaagattt gctgctacaa gaacttgttg ccacaaggtg ctccatcttc tccaaaattg
     6481 gctaacttga tttgctccaa gttggactac agaattcaag gttatgctgg ttctagaggt
     6541 ttgatataca ctagatacgc tgatgacttg accttgtctg ctcaatctat gaagaaagtt
     6601 gttaaggcca gagacttctt gttctccatt attccatcag aaggtttggt catcaactcc
     6661 aaaaagacct gtatttctgg tccaagatcc caaagaaaag ttaccggttt ggttatctcc
     6721 caagaaaagg ttggtattgg tagagaaaag tacaaagaaa tcagagccaa gatccaccat
     6781 attttctgcg gtaaatcctc cgaaatcgaa catgttagag gttggttgtc tttcatcttg
     6841 tccgttgatt ctaagtccca cagaagattg attacctaca tctctaagtt ggaaaagaag
     6901 tacggtaaga acccattgaa caaggctaag actccaccaa agaagaaaag aaaggtctga
     6961 tcccggttag taatgagttt aaacggggga ggctaactga aacacggaag gagacaatac
     7021 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac gggtgttggg
     7081 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     7141 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     7201 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcagat
     7261 ctgcgaaatc ggatccagct gaatctaagt ggatctgcga tcgctccggt gcccgtcagt
     7321 gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa
     7381 cgggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc
     7441 gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc
     7501 tttttcgcaa cgggtttgcc gccagaacac agctgaagct tcgaggggct cgcatctctc
     7561 cttcacgcgc ccgccgccct acctgaggcc gccatccacg ccggttgagt cgcgttctgc
     7621 cgcctcccgc ctgtggtgcc tcctgaactg cgtccgccgt ctaggtaagt ttaaagctca
     7681 ggtcgagacc gggcctttgt ccggcgctcc cttggagcct acctagactc agccggctct
     7741 ccacgctttg cctgaccctg cttgctcaac tctacgtctt tgtttcgttt tctgttctgc
     7801 gccgttacag atctaagctg tgaccggcgc ctactctaga gccgccatgt ctagactgga
     7861 caagagcaaa gtcataaact ctgctctgga attactcaat ggagtcggta tcgaaggcct
     7921 gacgacaagg aaactcgctc aaaagctggg agttgagcag cctaccctgt actggcacgt
     7981 gaagaacaag cgggccctgc tcgatgccct gccaatcgag atgctggaca ggcatcatac
     8041 ccactcctgc cccctggaag gcgagtcatg gcaagacttt ctgcggaaca acgccaagtc
     8101 ataccgctgt gctctcctct cacatcgcga cggggctaaa gtgcatctcg gcacccgccc
     8161 aacagagaaa cagtacgaaa ccctggaaaa tcagctcgcg ttcctgtgtc agcaaggctt
     8221 ctccctggag aacgcactgt acgctctgtc cgccgtgggc cactttacac tgggctgcgt
     8281 attggaggaa caggagcatc aagtagcaaa agaggaaaga gagacaccta ccaccgattc
     8341 tatgccccca cttctgaaac aagcaattga gctgttcgac cggcagggag ccgaacctgc
     8401 cttccttttc ggcctggaac taatcatatg tggcctggag aaacagctaa agtgcgaaag
     8461 cggcgggccg accgacgccc ttgacgattt tgacttagac atgctcccag ccgatgccct
     8521 tgacgacttt gaccttgata tgctgcctgc tgacgctctt gacgattttg accttgacat
     8581 gctccccggg gagggcagag gaagtcttct aacatgcggt gacgtggagg agaatcccgg
     8641 ccctatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc
     8701 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc
     8761 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct
     8821 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc
     8881 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag
     8941 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa
     9001 ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct
     9061 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt
     9121 cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac
     9181 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc
     9241 ctgaaatcaa cctctggatt acaaaatttg tgggtaaact tgtttattgc agcttataat
     9301 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     9361 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggaa ttgactcaaa
     9421 tgatgtcaat tagtctatca gaagctcatc tggtctccct tccgggggac aagacatccc
     9481 tgtttaatat ttaaacagca gtgttcccaa actgggttct tatatccctt gctctggtca
     9541 accaggttgc agggtttcct gtcctcacag gaacgaagtc cctaaagaaa cagtggcagc
     9601 caggtttagc cccggaattg actggattcc ttttttaggg cccattggta tggctttttc
     9661 cccgtatccc cccaggtgtc tgcaggctca aagagcagcg agaagcgttc agaggaaagc
     9721 gatcccgtgc caccttcccc gtgcccgggc tgtccccgca cgctgccggc tcggggatgc
     9781 ggggggagcg ccggaccgga gcggagcccc gggcggctcg ctgctgcccc ctagcggggg
     9841 agggacgtaa ttacatccct gggggctttg ggggggggct gtccctgata tctataacaa
     9901 gaaaatatat atataataag ttatcacgta agtagaacat gaaataacaa tataattatc
     9961 gtatgagtta aatcttaaaa gtcacgtaaa agataatcat gcgtcatttt gactcacgcg
    10021 gtcgttatag ttcaaaatca gtgacactta ccgcattgac aagcacgcct cacgggagct
    10081 ccaagcggcg actgagatgt cctaaatgca cagcgacgga ttcgcgctat ttagaaagag
    10141 agagcaatat ttcaagaatg catgcgtcaa ttttacgcag actatctttc tagggttaat
    10201 ctagctgcat caggatcata tcgtcgggtc ttttttccgg ctcagtcatc gcccaagctg
    10261 gcgctatctg ggcatcgggg aggaagaagc ccgtgccttt tcccgcgagg ttgaagcggc
    10321 atggaaagag tttgccgagg atgactgctg ctgcattgac gttgagcgaa aacgcacgtt
    10381 taccatgatg attcgggaag gtgtggccat gcacgccttt aacggtgaac tgttcgttca
    10441 ggccacctgg gataccagtt cgtcgcggct tttccggaca cagttccgga tggtcagccc
    10501 gaagcgcatc agcaacccga acaataccgg cgacagccgg aactgccgtg ccggtgtgca
    10561 gattaatgac agcggtgcgg cgctgggata ttacgtcagc gaggacgggt atcctggctg
    10621 gatgccgcag aaatggacat ggataccccg tgagttaccc ggcgggcgcg cttggcgtaa
    10681 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
    10741 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
    10801 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
    10861 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
    10921 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
    10981 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
    11041 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
    11101 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
    11161 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
    11221 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
    11281 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
    11341 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
    11401 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
    11461 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
    11521 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
    11581 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
    11641 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
    11701 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
    11761 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
    11821 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
    11881 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
    11941 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
    12001 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
    12061 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
    12121 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
    12181 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
    12241 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
    12301 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
    12361 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
    12421 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
    12481 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
    12541 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
    12601 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
    12661 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcat
//
