LOCUS       sequence-173280-       10896 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for genomic targeting of a Tetracycline-inducible Cas9 
            cassette to the human AAVS1/PPP1R12C locus with hygromycin 
            selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10896)
  AUTHORS   Shi ZD, Lee K, Yang D, Amin S, Verma N, Li QV, Zhu Z, Soh CL, Kumar 
            R, Evans T, Chen S, Huangfu D
  TITLE     Genome Editing in hPSCs Reveals GATA6 Haploinsufficiency and a 
            Genetic Interaction with GATA4 in Human Pancreatic Development.
  JOURNAL   Cell Stem Cell. 2017 Feb 8. pii: S1934-5909(17)30001-2. doi: 
            10.1016/j.stem.2017.01.001.
  PUBMED    28196600
REFERENCE   2  (bases 1 to 10896)
  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"; journalName: "Cell Stem 
            Cell. 2017 Feb 8. pii: S1934-5909(17)30001-2. doi: 
            10.1016/j.stem.2017.01.001."
COMMENT     Sequence Label: Hygro-Cas9 donor
FEATURES             Location/Qualifiers
     source          1..10896
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        24..128
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(129..197,198..989)
                     /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(347..366)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1160..1748
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1649..1668
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1902..1919
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2036..2057
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2072..2077,2078..2095,2096..2102)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2110..2126
                     /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     2115..2137
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2134..2150
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2134..2150
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2169..2189
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2171..2189
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    2208..3044
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     promoter        3075..3389
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    3083..3101
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3119..3137
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3154..3172
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3190..3208
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3226..3244
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3261..3279
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3297..3315
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3362..3386
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    3406..3430
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      3455..3464
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3464..3529
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3536..3556
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3581..7681
                     /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             3581..7681
                     /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             7682..7729
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     protein_bind    complement(7755..7779)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    7941..8022
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7978..7997)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    8029..8253
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     8242..8259
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(8291..9316)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(8291..9316)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(9326..9379)
                     /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"
     misc_feature    9403..9428
                     /label=SA
                     /note="splice acceptor site"
     misc_feature    9435..10238
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     primer_bind     complement(10252..10271)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(10253..10271)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(10281..10298)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(10281..10297)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10290..10312)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      10439..10894
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10526..10545)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10736..10757
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
       61 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
      121 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
      181 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
      241 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
      301 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
      361 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
      421 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
      481 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
      541 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
      601 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
      661 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
      721 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
      781 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
      841 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
      901 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
      961 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     1021 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     1081 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     1141 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     1201 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     1261 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     1321 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     1381 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     1441 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     1501 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     1561 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     1621 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     1681 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     1741 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     1801 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     1861 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     1921 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     1981 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     2041 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     2101 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     2161 ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggagctcaga gcagagccag
     2221 gaacccctgt agggaagggg caggagagcc aggggcatga gatggtggac gaggaagggg
     2281 gacagggaag cctgagcgcc tctcctgggc ttgccaagga ctcaaaccca gaagcccaga
     2341 gcagggcctt agggaagcgg gaccctgctc tgggcggagg aatatgtccc agatagcact
     2401 ggggactctt taaggaaaga aggatggaga aagagaaagg gagtagaggc ggccacgacc
     2461 tggtgaacac ctaggacgca ccattctcac aaagggagtt ttccacacgg acacccccct
     2521 cctcaccaca gccctgccag gacggggctg gctactggcc ttatctcaca ggtaaaactg
     2581 acgcacggag gaacaatata aattggggac tagaaaggtg aagagccaaa gttagaactc
     2641 aggaccaact tattctgatt ttgtttttcc aaactgcttc tcctcttggg aagtgtaagg
     2701 aagctgcagc accaggatca gtgaaacgca ccagacagcc gcgtcagagc agctcaggtt
     2761 ctgggagagg gtagcgcagg gtggccactg agaaccgggc aggtcacgca tccccccctt
     2821 ccctcccacc ccctgccaag ctctccctcc caggatcctc tctggctcca tcgtaagcaa
     2881 accttagagg ttctggcaag gagagagatg gctccaggaa atgggggtgt gtcaccagat
     2941 aaggaatctg cctaacagga ggtgggggtt agacccaata tcaggagact aggaaggagg
     3001 aggcctaagg atggggcttt tctgtcacca atcctgtccc tagtttaatt aaccctctag
     3061 agcttagtac tgtcgagttt actccctatc agtgatagag aacgtatgtc gagtttactc
     3121 cctatcagtg atagagaacg atgtcgagtt tactccctat cagtgataga gaacgtatgt
     3181 cgagtttact ccctatcagt gatagagaac gtatgtcgag tttactccct atcagtgata
     3241 gagaacgtat gtcgagttta tccctatcag tgatagagaa cgtatgtcga gtttactccc
     3301 tatcagtgat agagaacgta tgtcgaggta ggcgtgtacg gtgggaggcc tatataagca
     3361 gagctcgttt agtgaaccgt cagatcgcct ggagaattcc gatcaacaag tttgtacaaa
     3421 aaagcaggct ccgcggtttt caggttggac cggtgccacc atggactata aggaccacga
     3481 cggagactac aaggatcatg atattgatta caaagacgat gacgataaga tggccccaaa
     3541 gaagaagcgg aaggtcggta tccacggagt cccagcagcc gacaagaagt acagcatcgg
     3601 cctggacatc ggcaccaact ctgtgggctg ggccgtgatc accgacgagt acaaggtgcc
     3661 cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac agcatcaaga agaacctgat
     3721 cggagccctg ctgttcgaca gcggcgaaac agccgaggcc acccggctga agagaaccgc
     3781 cagaagaaga tacaccagac ggaagaaccg gatctgctat ctgcaagaga tcttcagcaa
     3841 cgagatggcc aaggtggacg acagcttctt ccacagactg gaagagtcct tcctggtgga
     3901 agaggataag aagcacgagc ggcaccccat cttcggcaac atcgtggacg aggtggccta
     3961 ccacgagaag taccccacca tctaccacct gagaaagaaa ctggtggaca gcaccgacaa
     4021 ggccgacctg cggctgatct atctggccct ggcccacatg atcaagttcc ggggccactt
     4081 cctgatcgag ggcgacctga accccgacaa cagcgacgtg gacaagctgt tcatccagct
     4141 ggtgcagacc tacaaccagc tgttcgagga aaaccccatc aacgccagcg gcgtggacgc
     4201 caaggccatc ctgtctgcca gactgagcaa gagcagacgg ctggaaaatc tgatcgccca
     4261 gctgcccggc gagaagaaga atggcctgtt cggaaacctg attgccctga gcctgggcct
     4321 gacccccaac ttcaagagca acttcgacct ggccgaggat gccaaactgc agctgagcaa
     4381 ggacacctac gacgacgacc tggacaacct gctggcccag atcggcgacc agtacgccga
     4441 cctgtttctg gccgccaaga acctgtccga cgccatcctg ctgagcgaca tcctgagagt
     4501 gaacaccgag atcaccaagg cccccctgag cgcctctatg atcaagagat acgacgagca
     4561 ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag cagctgcctg agaagtacaa
     4621 agagattttc ttcgaccaga gcaagaacgg ctacgccggc tacattgacg gcggagccag
     4681 ccaggaagag ttctacaagt tcatcaagcc catcctggaa aagatggacg gcaccgagga
     4741 actgctcgtg aagctgaaca gagaggacct gctgcggaag cagcggacct tcgacaacgg
     4801 cagcatcccc caccagatcc acctgggaga gctgcacgcc attctgcggc ggcaggaaga
     4861 tttttaccca ttcctgaagg acaaccggga aaagatcgag aagatcctga ccttccgcat
     4921 cccctactac gtgggccctc tggccagggg aaacagcaga ttcgcctgga tgaccagaaa
     4981 gagcgaggaa accatcaccc cctggaactt cgaggaagtg gtggacaagg gcgcttccgc
     5041 ccagagcttc atcgagcgga tgaccaactt cgataagaac ctgcccaacg agaaggtgct
     5101 gcccaagcac agcctgctgt acgagtactt caccgtgtat aacgagctga ccaaagtgaa
     5161 atacgtgacc gagggaatga gaaagcccgc cttcctgagc ggcgagcaga aaaaggccat
     5221 cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg aagcagctga aagaggacta
     5281 cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc ggcgtggaag atcggttcaa
     5341 cgcctccctg ggcacatacc acgatctgct gaaaattatc aaggacaagg acttcctgga
     5401 caatgaggaa aacgaggaca ttctggaaga tatcgtgctg accctgacac tgtttgagga
     5461 cagagagatg atcgaggaac ggctgaaaac ctatgcccac ctgttcgacg acaaagtgat
     5521 gaagcagctg aagcggcgga gatacaccgg ctggggcagg ctgagccgga agctgatcaa
     5581 cggcatccgg gacaagcagt ccggcaagac aatcctggat ttcctgaagt ccgacggctt
     5641 cgccaacaga aacttcatgc agctgatcca cgacgacagc ctgaccttta aagaggacat
     5701 ccagaaagcc caggtgtccg gccagggcga tagcctgcac gagcacattg ccaatctggc
     5761 cggcagcccc gccattaaga agggcatcct gcagacagtg aaggtggtgg acgagctcgt
     5821 gaaagtgatg ggccggcaca agcccgagaa catcgtgatc gaaatggcca gagagaacca
     5881 gaccacccag aagggacaga agaacagccg cgagagaatg aagcggatcg aagagggcat
     5941 caaagagctg ggcagccaga tcctgaaaga acaccccgtg gaaaacaccc agctgcagaa
     6001 cgagaagctg tacctgtact acctgcagaa tgggcgggat atgtacgtgg accaggaact
     6061 ggacatcaac cggctgtccg actacgatgt ggaccatatc gtgcctcaga gctttctgaa
     6121 ggacgactcc atcgacaaca aggtgctgac cagaagcgac aagaaccggg gcaagagcga
     6181 caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac tactggcggc agctgctgaa
     6241 cgccaagctg attacccaga gaaagttcga caatctgacc aaggccgaga gaggcggcct
     6301 gagcgaactg gataaggccg gcttcatcaa gagacagctg gtggaaaccc ggcagatcac
     6361 aaagcacgtg gcacagatcc tggactcccg gatgaacact aagtacgacg agaatgacaa
     6421 gctgatccgg gaagtgaaag tgatcaccct gaagtccaag ctggtgtccg atttccggaa
     6481 ggatttccag ttttacaaag tgcgcgagat caacaactac caccacgccc acgacgccta
     6541 cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac cctaagctgg aaagcgagtt
     6601 cgtgtacggc gactacaagg tgtacgacgt gcggaagatg atcgccaaga gcgagcagga
     6661 aatcggcaag gctaccgcca agtacttctt ctacagcaac atcatgaact ttttcaagac
     6721 cgagattacc ctggccaacg gcgagatccg gaagcggcct ctgatcgaga caaacggcga
     6781 aaccggggag atcgtgtggg ataagggccg ggattttgcc accgtgcgga aagtgctgag
     6841 catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag acaggcggct tcagcaaaga
     6901 gtctatcctg cccaagagga acagcgataa gctgatcgcc agaaagaagg actgggaccc
     6961 taagaagtac ggcggcttcg acagccccac cgtggcctat tctgtgctgg tggtggccaa
     7021 agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa gagctgctgg ggatcaccat
     7081 catggaaaga agcagcttcg agaagaatcc catcgacttt ctggaagcca agggctacaa
     7141 agaagtgaaa aaggacctga tcatcaagct gcctaagtac tccctgttcg agctggaaaa
     7201 cggccggaag agaatgctgg cctctgccgg cgaactgcag aagggaaacg aactggccct
     7261 gccctccaaa tatgtgaact tcctgtacct ggccagccac tatgagaagc tgaagggctc
     7321 ccccgaggat aatgagcaga aacagctgtt tgtggaacag cacaagcact acctggacga
     7381 gatcatcgag cagatcagcg agttctccaa gagagtgatc ctggccgacg ctaatctgga
     7441 caaagtgctg tccgcctaca acaagcaccg ggataagccc atcagagagc aggccgagaa
     7501 tatcatccac ctgtttaccc tgaccaatct gggagcccct gccgccttca agtactttga
     7561 caccaccatc gaccggaaga ggtacaccag caccaaagag gtgctggacg ccaccctgat
     7621 ccaccagagc atcaccggcc tgtacgagac acggatcgac ctgtctcagc tgggaggcga
     7681 caaaaggccg gcggccacga aaaaggccgg ccaggcaaaa aagaaaaagt aagaattcaa
     7741 gggtgggcgc gccgacccag ctttcttgta caaagtggtt gatccgggga tcctctagtc
     7801 agcgtttatc tgcagaattc gcccttgttt aaacgatcat aatcagccat accacatttg
     7861 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     7921 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     7981 atagcatcac aaatttcaca aataaagcat ttttttcact gcctcgaccc atagagccca
     8041 ccgcatcccc agcatgcctg ctattgtctt cccaatcctc ccccttgctg tcctgcccca
     8101 ccccaccccc cagaatagaa tgacacctac tcagacaatg cgatgcaatt tcctcatttt
     8161 attaggaaag gacagtggga gtggcacctt ccagggtcaa ggaaggcacg ggggaggggc
     8221 aaacaacaga tggctggcaa ctagaaggca cagtcgaggc tgatcagcgg gtttaaacgg
     8281 gccctctaga ctattccttt gccctcggac gagtgctggg gcgtcggttt ccactatcgg
     8341 cgagtacttc tacacagcca tcggtccaga cggccgcgct tctgcgggcg atttgtgtac
     8401 gcccgacagt cccggctccg gatcggacga ttgcgtcgca tcgaccctgc gcccaagctg
     8461 catcatcgaa attgccgtca accaagctct gatagagttg gtcaagacca atgcggagca
     8521 tatacgcccg gagccgcggc gatcctgcaa gctccggatg cctccgctcg aagtagcgcg
     8581 tctgctgctc catacaagcc aaccacggcc tccagaagaa gatgttggcg acctcgtatt
     8641 gggaatcccc gaacatcgcc tcgctccagt caatgaccgc tgttatgcgg ccattgtccg
     8701 tcaggacatt gttggagccg aaatccgcgt gcacgaggtg ccggacttcg gggcagtcct
     8761 cggcccaaag catcagctca tcgagagcct gcgcgacgga cgcactgacg gtgtcgtcca
     8821 tcacagtttg ccagtgatac acatggggat cagcaatcgc gcatatgaaa tcacgccatg
     8881 tagtgtattg accgattcct tgcggtccga atgggccgaa cccgctcgtc tggctaagat
     8941 cggccgcagc gatcgcatcc atggcctccg cgaccggctg cagaacagcg ggcagttcgg
     9001 tttcaggcag gtcttgcaac gtgacaccct gtgcacggcg ggagatgcaa taggtcaggc
     9061 tctcgctgaa ttccccaatg tcaagcactt ccggaatcgg gagcgcggcc gatgcaaagt
     9121 gccgataaac ataacgatct ttgtagaaac catcggcgca gctatttacc cgcaggacat
     9181 atccacgccc tcctacatcg aagctgaaag cacgagattc ttcgccctcc gagagctgca
     9241 tcaggtcgga gacgctgtcg aacttttcga tcagaaactt ctcgacagac gtcgcggtga
     9301 gttcaggctt tttcatctgc agcctagggc cgggattctc ctccacgtca ccgcatgtta
     9361 gaagacttcc tctgccctct ccgctgccag atctctcgag gccctgtggg aggaagagaa
     9421 gaggtcaggg atctgcccca ctgtggggtg gaggggacag ataaaagtac ccagaaccag
     9481 agccacatta accggccctg ggaatataag gtggtcccag ctcggggaca caggatccct
     9541 ggaggcagca aacatgctgt cctgaagtgg acataggggc ccgggttgga ggaagaagac
     9601 tagctgagct ctcggacccc tggaagatgc catgacaggg ggctggaaga gctagcacag
     9661 actagagagg taaggggggt aggggagctg cccaaatgaa aggagtgaga ggtgacccga
     9721 atccacagga gaacggggtg tccaggcaaa gaaagcaaga ggatggagag gtggctaaag
     9781 ccagggagac ggggtacttt ggggttgtcc agaaaaacgg tgatgatgca ggcctacaag
     9841 aaggggaggc gggacgcaag ggagacatcc gtcggagaag gccatcctaa gaaacgagag
     9901 atggcacagg ccccagaagg agaaggaaaa gggaacccag cgagtgaaga cggcatgggg
     9961 ttgggtgagg gaggagagat gcccggagag gacccagaca cggggaggat ccgctcagag
    10021 gacatcacgt ggtgcagcgc cgagaaggaa gtgctccgga aagagcatcc ttgggcagca
    10081 acacagcaga gagcaagggg aagagggagt ggaggaagac ggaacctgaa ggaggcggca
    10141 gggaaggatc tgggccagcc gtagaggtga cccaggccac aagctgcaga cagaaagcgg
    10201 cacaggccca ggggagagaa tgctggtcag agaaagcagg tacccaattc gccctatagt
    10261 gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
    10321 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
    10381 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac
    10441 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
    10501 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
    10561 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
    10621 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
    10681 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
    10741 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
    10801 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
    10861 gcgaatttta acaaaatatt aacgcttaca atttag
//
