LOCUS       sequence-374352-       11708 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid containing Cas9 and ANKRD1 sgRNA2 , sgRNA sequence: 
            cggtcagcttatatagct..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11708)
  AUTHORS   Meng Q, Stoyko D, Andrews CM, Konstantinidou P, Genzor P, O T, 
            Elchert AR, Benner L, Sobti S, Katz EY, Haase AD
  TITLE     Functional editing of endogenous genes through rapid selection of 
            cell pools (Rapid generation of endogenously tagged genes in 
            Drosophila ovarian somatic sheath cells).
  JOURNAL   Nucleic Acids Res. 2022 May 27. pii: 6594084. doi: 
            10.1093/nar/gkac448.
  PUBMED    35639929
REFERENCE   2  (bases 1 to 11708)
  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: "Nucleic 
            Acids Res. 2022 May 27. pii: 6594084. doi: 10.1093/nar/gkac448."
COMMENT     Sequence Label: Cas9_ANKRD1_sgRNA2
FEATURES             Location/Qualifiers
     source          1..11708
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     331..350
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     584..601
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    718..739
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(754..759,760..777,778..784)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    792..808
                     /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     797..819
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     816..832
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     816..832
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     851..871
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        853..871
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        899..1125
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             1126..1306
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1353..1478
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1971..2204
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2389..2433
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     promoter        2611..2859
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2611..2631
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2782..2801
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2879..2954
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    3002..3119
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3188..3399
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      3419..3428
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3428..3451
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3458..3478
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3503..7603
                     /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             3503..7603
                     /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             7604..7651
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7667..7723
                     /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             join(7724..7726,7727..7729,7730..8440)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(8030..8049)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8377..8398
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    8459..9047
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8512..8532)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8930..8941)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9119..9352
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    9430..9551
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9467..9486)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9521..9540
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      9591..9726
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(9746..9765)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9747..9765)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9775..9792)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9775..9791)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9784..9806)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9933..10388
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10020..10039)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10230..10251
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        10414..10518
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10519..10587,10588..11379)
                     /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(10737..10756)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(11550..11708,1..430)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
       61 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
      121 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
      181 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
      241 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
      301 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
      361 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
      421 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
      481 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
      541 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
      601 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
      661 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
      721 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
      781 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
      841 cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctgc aagcttaatg
      901 tagtcttatg caatactctt gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc
      961 ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt
     1021 gccttattag gaaggcaaca gacgggtctg acatggattg gacgaaccac tgaattgccg
     1081 cattgcagag atattgtatt taagtgccta gctcgataca taaacgggtc tctctggtta
     1141 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     1201 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     1261 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca
     1321 gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg
     1381 aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta
     1441 gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta
     1501 gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac
     1561 atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa
     1621 catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag
     1681 aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag
     1741 agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga
     1801 ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat
     1861 tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc
     1921 accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg
     1981 ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg
     2041 gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct
     2101 attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca
     2161 agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc
     2221 tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct
     2281 ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac
     2341 acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa
     2401 gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg
     2461 ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt
     2521 tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag
     2581 acccacctcc caaccccgag gggacccaga gagggcctat ttcccatgat tccttcatat
     2641 ttgcatatac gatacaaggc tgttagagag ataattagaa ttaatttgac tgtaaacaca
     2701 aagatattag tacaaaatac gtgacgtaga aagtaataat ttcttgggta gtttgcagtt
     2761 ttaaaattat gttttaaaat ggactatcat atgcttaccg taacttgaaa gtatttcgat
     2821 ttcttggctt tatatatctt gtggaaagga cgaaacaccg cggtcagctt atatagctgt
     2881 tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg
     2941 caccgagtcg gtgctttttt gaattctaga tcttgagaca aatggcagta ttcatccaca
     3001 attttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa
     3061 tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc
     3121 gggtttatta cagggacagc agagatccac tttggcgccg gctcgagtgg ctccggtgcc
     3181 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc
     3241 aattgaaccg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac
     3301 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg
     3361 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg tgtcgtgacg cgggatccgc
     3421 caccatggat tacaaagacg atgacgataa gatggcccca aagaagaagc ggaaggtcgg
     3481 tatccacgga gtcccagcag ccgacaagaa gtacagcatc ggcctggaca tcggcaccaa
     3541 ctctgtgggc tgggccgtga tcaccgacga gtacaaggtg cccagcaaga aattcaaggt
     3601 gctgggcaac accgaccggc acagcatcaa gaagaacctg atcggagccc tgctgttcga
     3661 cagcggcgaa acagccgagg ccacccggct gaagagaacc gccagaagaa gatacaccag
     3721 acggaagaac cggatctgct atctgcaaga gatcttcagc aacgagatgg ccaaggtgga
     3781 cgacagcttc ttccacagac tggaagagtc cttcctggtg gaagaggata agaagcacga
     3841 gcggcacccc atcttcggca acatcgtgga cgaggtggcc taccacgaga agtaccccac
     3901 catctaccac ctgagaaaga aactggtgga cagcaccgac aaggccgacc tgcggctgat
     3961 ctatctggcc ctggcccaca tgatcaagtt ccggggccac ttcctgatcg agggcgacct
     4021 gaaccccgac aacagcgacg tggacaagct gttcatccag ctggtgcaga cctacaacca
     4081 gctgttcgag gaaaacccca tcaacgccag cggcgtggac gccaaggcca tcctgtctgc
     4141 cagactgagc aagagcagac ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa
     4201 gaatggcctg ttcggaaacc tgattgccct gagcctgggc ctgaccccca acttcaagag
     4261 caacttcgac ctggccgagg atgccaaact gcagctgagc aaggacacct acgacgacga
     4321 cctggacaac ctgctggccc agatcggcga ccagtacgcc gacctgtttc tggccgccaa
     4381 gaacctgtcc gacgccatcc tgctgagcga catcctgaga gtgaacaccg agatcaccaa
     4441 ggcccccctg agcgcctcta tgatcaagag atacgacgag caccaccagg acctgaccct
     4501 gctgaaagct ctcgtgcggc agcagctgcc tgagaagtac aaagagattt tcttcgacca
     4561 gagcaagaac ggctacgccg gctacattga cggcggagcc agccaggaag agttctacaa
     4621 gttcatcaag cccatcctgg aaaagatgga cggcaccgag gaactgctcg tgaagctgaa
     4681 cagagaggac ctgctgcgga agcagcggac cttcgacaac ggcagcatcc cccaccagat
     4741 ccacctggga gagctgcacg ccattctgcg gcggcaggaa gatttttacc cattcctgaa
     4801 ggacaaccgg gaaaagatcg agaagatcct gaccttccgc atcccctact acgtgggccc
     4861 tctggccagg ggaaacagca gattcgcctg gatgaccaga aagagcgagg aaaccatcac
     4921 cccctggaac ttcgaggaag tggtggacaa gggcgcttcc gcccagagct tcatcgagcg
     4981 gatgaccaac ttcgataaga acctgcccaa cgagaaggtg ctgcccaagc acagcctgct
     5041 gtacgagtac ttcaccgtgt ataacgagct gaccaaagtg aaatacgtga ccgagggaat
     5101 gagaaagccc gccttcctga gcggcgagca gaaaaaggcc atcgtggacc tgctgttcaa
     5161 gaccaaccgg aaagtgaccg tgaagcagct gaaagaggac tacttcaaga aaatcgagtg
     5221 cttcgactcc gtggaaatct ccggcgtgga agatcggttc aacgcctccc tgggcacata
     5281 ccacgatctg ctgaaaatta tcaaggacaa ggacttcctg gacaatgagg aaaacgagga
     5341 cattctggaa gatatcgtgc tgaccctgac actgtttgag gacagagaga tgatcgagga
     5401 acggctgaaa acctatgccc acctgttcga cgacaaagtg atgaagcagc tgaagcggcg
     5461 gagatacacc ggctggggca ggctgagccg gaagctgatc aacggcatcc gggacaagca
     5521 gtccggcaag acaatcctgg atttcctgaa gtccgacggc ttcgccaaca gaaacttcat
     5581 gcagctgatc cacgacgaca gcctgacctt taaagaggac atccagaaag cccaggtgtc
     5641 cggccagggc gatagcctgc acgagcacat tgccaatctg gccggcagcc ccgccattaa
     5701 gaagggcatc ctgcagacag tgaaggtggt ggacgagctc gtgaaagtga tgggccggca
     5761 caagcccgag aacatcgtga tcgaaatggc cagagagaac cagaccaccc agaagggaca
     5821 gaagaacagc cgcgagagaa tgaagcggat cgaagagggc atcaaagagc tgggcagcca
     5881 gatcctgaaa gaacaccccg tggaaaacac ccagctgcag aacgagaagc tgtacctgta
     5941 ctacctgcag aatgggcggg atatgtacgt ggaccaggaa ctggacatca accggctgtc
     6001 cgactacgat gtggaccata tcgtgcctca gagctttctg aaggacgact ccatcgacaa
     6061 caaggtgctg accagaagcg acaagaaccg gggcaagagc gacaacgtgc cctccgaaga
     6121 ggtcgtgaag aagatgaaga actactggcg gcagctgctg aacgccaagc tgattaccca
     6181 gagaaagttc gacaatctga ccaaggccga gagaggcggc ctgagcgaac tggataaggc
     6241 cggcttcatc aagagacagc tggtggaaac ccggcagatc acaaagcacg tggcacagat
     6301 cctggactcc cggatgaaca ctaagtacga cgagaatgac aagctgatcc gggaagtgaa
     6361 agtgatcacc ctgaagtcca agctggtgtc cgatttccgg aaggatttcc agttttacaa
     6421 agtgcgcgag atcaacaact accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg
     6481 aaccgccctg atcaaaaagt accctaagct ggaaagcgag ttcgtgtacg gcgactacaa
     6541 ggtgtacgac gtgcggaaga tgatcgccaa gagcgagcag gaaatcggca aggctaccgc
     6601 caagtacttc ttctacagca acatcatgaa ctttttcaag accgagatta ccctggccaa
     6661 cggcgagatc cggaagcggc ctctgatcga gacaaacggc gaaaccgggg agatcgtgtg
     6721 ggataagggc cgggattttg ccaccgtgcg gaaagtgctg agcatgcccc aagtgaatat
     6781 cgtgaaaaag accgaggtgc agacaggcgg cttcagcaaa gagtctatcc tgcccaagag
     6841 gaacagcgat aagctgatcg ccagaaagaa ggactgggac cctaagaagt acggcggctt
     6901 cgacagcccc accgtggcct attctgtgct ggtggtggcc aaagtggaaa agggcaagtc
     6961 caagaaactg aagagtgtga aagagctgct ggggatcacc atcatggaaa gaagcagctt
     7021 cgagaagaat cccatcgact ttctggaagc caagggctac aaagaagtga aaaaggacct
     7081 gatcatcaag ctgcctaagt actccctgtt cgagctggaa aacggccgga agagaatgct
     7141 ggcctctgcc ggcgaactgc agaagggaaa cgaactggcc ctgccctcca aatatgtgaa
     7201 cttcctgtac ctggccagcc actatgagaa gctgaagggc tcccccgagg ataatgagca
     7261 gaaacagctg tttgtggaac agcacaagca ctacctggac gagatcatcg agcagatcag
     7321 cgagttctcc aagagagtga tcctggccga cgctaatctg gacaaagtgc tgtccgccta
     7381 caacaagcac cgggataagc ccatcagaga gcaggccgag aatatcatcc acctgtttac
     7441 cctgaccaat ctgggagccc ctgccgcctt caagtacttt gacaccacca tcgaccggaa
     7501 gaggtacacc agcaccaaag aggtgctgga cgccaccctg atccaccaga gcatcaccgg
     7561 cctgtacgag acacggatcg acctgtctca gctgggaggc gacaagcgtc ctgctgctac
     7621 taagaaagct ggtcaagcta agaaaaagaa agctagcggc agcggcgcca ccaacttcag
     7681 cctgctgaag caggccggcg acgtggagga gaaccccggc cccatggtgt ctaagggcga
     7741 agagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     7801 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     7861 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     7921 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     7981 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     8041 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     8101 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     8161 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
     8221 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     8281 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     8341 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     8401 cgccgccggg atcactctcg gcatggacga gctgtacaag taaacgcgtt aagtcgacaa
     8461 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     8521 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     8581 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     8641 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     8701 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     8761 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     8821 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
     8881 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     8941 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     9001 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcgtc gactttaaga
     9061 ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg
     9121 gaagggctaa ttcactccca acgaagacaa gatctgcttt ttgcttgtac tgggtctctc
     9181 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
     9241 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
     9301 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtacgtat
     9361 agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag
     9421 agtgagagga acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     9481 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     9541 aatgtatctt atcatgtctg gctctagcta tcccgcccct aactccgccc atcccgcccc
     9601 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     9661 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     9721 gaggcctagg gacgtaccca attcgcccta tagtgagtcg tattacgcgc gctcactggc
     9781 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     9841 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     9901 ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc
     9961 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
    10021 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
    10081 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
    10141 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
    10201 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
    10261 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
    10321 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
    10381 tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
    10441 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
    10501 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
    10561 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
    10621 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
    10681 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
    10741 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
    10801 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
    10861 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
    10921 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
    10981 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
    11041 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
    11101 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
    11161 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
    11221 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
    11281 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
    11341 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
    11401 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
    11461 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
    11521 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
    11581 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
    11641 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
    11701 ctagtgta
//
