LOCUS       sequence-420786-       13790 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Guide RNA targeting SERPINB3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13790)
  AUTHORS   Ohara Y, Tang W, Liu H, Yang S, Dorsey TH, Cawley H, Moreno P, Chari
            R, Guest MR, Azizian A, Gaedcke J, Ghadimi M, Hanna N, Ambs S, 
            Hussain SP
  TITLE     SERPINB3-MYC axis induces the basal-like/squamous subtype and 
            enhances disease progression in pancreatic cancer.
  JOURNAL   Cell Rep. 2023 Nov 18;42(12):113434. doi: 
            10.1016/j.celrep.2023.113434.
  PUBMED    37980563
REFERENCE   2  (bases 1 to 13790)
  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 Rep.";
            date: "2023-11-18"; volume: "42(12):113434. doi"; pages: " 
            10.1016/j.celrep.2023.113434"
COMMENT     Sequence Label: pGMC00036 (aka pMC0223)
FEATURES             Location/Qualifiers
     source          1..13790
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        34..232
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     187..207
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             250..430
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    477..602
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1095..1328
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1513..1557
                     /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"
     misc_feature    1855..1972
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2023..2271
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2023..2043
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2194..2213
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2293..2368
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2430..2641
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2660..2669
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2666..6769
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             2666..6769
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             6770..6817
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6818..6841
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             6851..6904
                     /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             join(6905..6907,6908..7618)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(6905..6907,6908..7618)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(7208..7227)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7555..7576
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             7628..7684
                     /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             7685..8281
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     8178..8198
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8297..8885
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8350..8370)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8768..8779)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8957..9190
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9216..9233)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9222..9446
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9492..9920
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9579..9598)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9789..9810
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9929..9949)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9934..10263
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10114..10249
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10176..10195
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10311..10358
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10377..10751
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    10881..11002
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10918..10937)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10972..10991
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11051..11067)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11051..11067)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11064..11086)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11075..11091
                     /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(11099..11105,11106..11123,11124..11129))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11144..11165
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11282..11299)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11453..12041)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11533..11552)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12212..13003,13004..13072))
                     /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     12835..12854
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13073..13177)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(13252..13271)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(13443..13790,1..32)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg
       61 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
      121 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
      181 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agcgcgtttt
      241 gcctgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
      301 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
      361 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
      421 atctctagca gtggcgcccg aacagggact tgaaagcgaa agggaaacca gaggagctct
      481 ctcgacgcag gactcggctt gctgaagcgc gcacggcaag aggcgagggg cggcgactgg
      541 tgagtacgcc aaaaattttg actagcggag gctagaagga gagagatggg tgcgagagcg
      601 tcagtattaa gcgggggaga attagatcgc gatgggaaaa aattcggtta aggccagggg
      661 gaaagaaaaa atataaatta aaacatatag tatgggcaag cagggagcta gaacgattcg
      721 cagttaatcc tggcctgtta gaaacatcag aaggctgtag acaaatactg ggacagctac
      781 aaccatccct tcagacagga tcagaagaac ttagatcatt atataataca gtagcaaccc
      841 tctattgtgt gcatcaaagg atagagataa aagacaccaa ggaagcttta gacaagatag
      901 aggaagagca aaacaaaagt aagaccaccg cacagcaagc ggccgctgat cttcagacct
      961 ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa
     1021 attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa
     1081 agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg
     1141 ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag
     1201 cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc
     1261 tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa
     1321 cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg
     1381 aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag
     1441 tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa
     1501 aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg
     1561 aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga
     1621 ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag
     1681 ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc
     1741 gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac
     1801 ggatcggcac tgcgtgcgcc aattctgcag acaaatggca gtattcatcc acaattttaa
     1861 aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac
     1921 agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta
     1981 ttacagggac agcagagatc cagtttggtt aattaaggta ccgagggcct atttcccatg
     2041 attccttcat atttgcatat acgatacaag gctgttagag agataattag aattaatttg
     2101 actgtaaaca caaagatatt agtacaaaat acgtgacgta gaaagtaata atttcttggg
     2161 tagtttgcag ttttaaaatt atgttttaaa atggactatc atatgcttac cgtaacttga
     2221 aagtatttcg atttcttggc tttatatatc ttgtggaaag gacgaaacac cgaaaatcaa
     2281 cagattccac acgttttaga gctagaaata gcaagttaaa ataaggctag tccgttatca
     2341 acttgaaaaa gtggcaccga gtcggtgctt ttttgaattc gctagctagg tcttgaaagg
     2401 agtgggaatt ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga
     2461 gaagttgggg ggaggggtcg gcaattgatc cggtgcctag agaaggtggc gcggggtaaa
     2521 ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta
     2581 tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca
     2641 ggaccggttc tagagcgctg ccaccatgga caagaagtac agcatcggcc tggacatcgg
     2701 caccaactct gtgggctggg ccgtgatcac cgacgagtac aaggtgccca gcaagaaatt
     2761 caaggtgctg ggcaacaccg accggcacag catcaagaag aacctgatcg gagccctgct
     2821 gttcgacagc ggcgaaacag ccgaggccac ccggctgaag agaaccgcca gaagaagata
     2881 caccagacgg aagaaccgga tctgctatct gcaagagatc ttcagcaacg agatggccaa
     2941 ggtggacgac agcttcttcc acagactgga agagtccttc ctggtggaag aggataagaa
     3001 gcacgagcgg caccccatct tcggcaacat cgtggacgag gtggcctacc acgagaagta
     3061 ccccaccatc taccacctga gaaagaaact ggtggacagc accgacaagg ccgacctgcg
     3121 gctgatctat ctggccctgg cccacatgat caagttccgg ggccacttcc tgatcgaggg
     3181 cgacctgaac cccgacaaca gcgacgtgga caagctgttc atccagctgg tgcagaccta
     3241 caaccagctg ttcgaggaaa accccatcaa cgccagcggc gtggacgcca aggccatcct
     3301 gtctgccaga ctgagcaaga gcagacggct ggaaaatctg atcgcccagc tgcccggcga
     3361 gaagaagaat ggcctgttcg gaaacctgat tgccctgagc ctgggcctga cccccaactt
     3421 caagagcaac ttcgacctgg ccgaggatgc caaactgcag ctgagcaagg acacctacga
     3481 cgacgacctg gacaacctgc tggcccagat cggcgaccag tacgccgacc tgtttctggc
     3541 cgccaagaac ctgtccgacg ccatcctgct gagcgacatc ctgagagtga acaccgagat
     3601 caccaaggcc cccctgagcg cctctatgat caagagatac gacgagcacc accaggacct
     3661 gaccctgctg aaagctctcg tgcggcagca gctgcctgag aagtacaaag agattttctt
     3721 cgaccagagc aagaacggct acgccggcta cattgacggc ggagccagcc aggaagagtt
     3781 ctacaagttc atcaagccca tcctggaaaa gatggacggc accgaggaac tgctcgtgaa
     3841 gctgaacaga gaggacctgc tgcggaagca gcggaccttc gacaacggca gcatccccca
     3901 ccagatccac ctgggagagc tgcacgccat tctgcggcgg caggaagatt tttacccatt
     3961 cctgaaggac aaccgggaaa agatcgagaa gatcctgacc ttccgcatcc cctactacgt
     4021 gggccctctg gccaggggaa acagcagatt cgcctggatg accagaaaga gcgaggaaac
     4081 catcaccccc tggaacttcg aggaagtggt ggacaagggc gcttccgccc agagcttcat
     4141 cgagcggatg accaacttcg ataagaacct gcccaacgag aaggtgctgc ccaagcacag
     4201 cctgctgtac gagtacttca ccgtgtataa cgagctgacc aaagtgaaat acgtgaccga
     4261 gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa aaggccatcg tggacctgct
     4321 gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa gaggactact tcaagaaaat
     4381 cgagtgcttc gactccgtgg aaatctccgg cgtggaagat cggttcaacg cctccctggg
     4441 cacataccac gatctgctga aaattatcaa ggacaaggac ttcctggaca atgaggaaaa
     4501 cgaggacatt ctggaagata tcgtgctgac cctgacactg tttgaggaca gagagatgat
     4561 cgaggaacgg ctgaaaacct atgcccacct gttcgacgac aaagtgatga agcagctgaa
     4621 gcggcggaga tacaccggct ggggcaggct gagccggaag ctgatcaacg gcatccggga
     4681 caagcagtcc ggcaagacaa tcctggattt cctgaagtcc gacggcttcg ccaacagaaa
     4741 cttcatgcag ctgatccacg acgacagcct gacctttaaa gaggacatcc agaaagccca
     4801 ggtgtccggc cagggcgata gcctgcacga gcacattgcc aatctggccg gcagccccgc
     4861 cattaagaag ggcatcctgc agacagtgaa ggtggtggac gagctcgtga aagtgatggg
     4921 ccggcacaag cccgagaaca tcgtgatcga aatggccaga gagaaccaga ccacccagaa
     4981 gggacagaag aacagccgcg agagaatgaa gcggatcgaa gagggcatca aagagctggg
     5041 cagccagatc ctgaaagaac accccgtgga aaacacccag ctgcagaacg agaagctgta
     5101 cctgtactac ctgcagaatg ggcgggatat gtacgtggac caggaactgg acatcaaccg
     5161 gctgtccgac tacgatgtgg accatatcgt gcctcagagc tttctgaagg acgactccat
     5221 cgacaacaag gtgctgacca gaagcgacaa gaaccggggc aagagcgaca acgtgccctc
     5281 cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag ctgctgaacg ccaagctgat
     5341 tacccagaga aagttcgaca atctgaccaa ggccgagaga ggcggcctga gcgaactgga
     5401 taaggccggc ttcatcaaga gacagctggt ggaaacccgg cagatcacaa agcacgtggc
     5461 acagatcctg gactcccgga tgaacactaa gtacgacgag aatgacaagc tgatccggga
     5521 agtgaaagtg atcaccctga agtccaagct ggtgtccgat ttccggaagg atttccagtt
     5581 ttacaaagtg cgcgagatca acaactacca ccacgcccac gacgcctacc tgaacgccgt
     5641 cgtgggaacc gccctgatca aaaagtaccc taagctggaa agcgagttcg tgtacggcga
     5701 ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa tcggcaaggc
     5761 taccgccaag tacttcttct acagcaacat catgaacttt ttcaagaccg agattaccct
     5821 ggccaacggc gagatccgga agcggcctct gatcgagaca aacggcgaaa ccggggagat
     5881 cgtgtgggat aagggccggg attttgccac cgtgcggaaa gtgctgagca tgccccaagt
     5941 gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc agcaaagagt ctatcctgcc
     6001 caagaggaac agcgataagc tgatcgccag aaagaaggac tgggacccta agaagtacgg
     6061 cggcttcgac agccccaccg tggcctattc tgtgctggtg gtggccaaag tggaaaaggg
     6121 caagtccaag aaactgaaga gtgtgaaaga gctgctgggg atcaccatca tggaaagaag
     6181 cagcttcgag aagaatccca tcgactttct ggaagccaag ggctacaaag aagtgaaaaa
     6241 ggacctgatc atcaagctgc ctaagtactc cctgttcgag ctggaaaacg gccggaagag
     6301 aatgctggcc tctgccggcg aactgcagaa gggaaacgaa ctggccctgc cctccaaata
     6361 tgtgaacttc ctgtacctgg ccagccacta tgagaagctg aagggctccc ccgaggataa
     6421 tgagcagaaa cagctgtttg tggaacagca caagcactac ctggacgaga tcatcgagca
     6481 gatcagcgag ttctccaaga gagtgatcct ggccgacgct aatctggaca aagtgctgtc
     6541 cgcctacaac aagcaccggg ataagcccat cagagagcag gccgagaata tcatccacct
     6601 gtttaccctg accaatctgg gagcccctgc cgccttcaag tactttgaca ccaccatcga
     6661 ccggaagagg tacaccagca ccaaagaggt gctggacgcc accctgatcc accagagcat
     6721 caccggcctg tacgagacac ggatcgacct gtctcagctg ggaggcgaca agcgacctgc
     6781 cgccacaaag aaggctggac aggctaagaa gaagaaagat tacaaagacg atgacgataa
     6841 gggcagtgga gagggcagag gaagtctgct aacatgcggt gacgtcgagg agaatcctgg
     6901 cccagtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     6961 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     7021 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     7081 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     7141 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     7201 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     7261 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     7321 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     7381 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     7441 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     7501 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     7561 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaaggg
     7621 atccggcgca acaaacttct ctctgctgaa acaagccgga gatgtcgaag agaatcctgg
     7681 accgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt
     7741 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga
     7801 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga
     7861 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga
     7921 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg
     7981 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga
     8041 gcccgcgtgg ttcctggcca ccgtcggagt ctcgcccgac caccagggca agggtctggg
     8101 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct
     8161 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc
     8221 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg
     8281 aacgcgttaa gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     8341 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     8401 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     8461 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     8521 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     8581 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     8641 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     8701 ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     8761 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     8821 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     8881 cccgcgtcga ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
     8941 aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt
     9001 gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
     9061 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
     9121 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
     9181 atctctagca gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     9241 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     9301 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     9361 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     9421 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     9481 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     9541 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     9601 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     9661 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     9721 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     9781 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     9841 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     9901 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     9961 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
    10021 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
    10081 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
    10141 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
    10201 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
    10261 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcagcacgt gttgacaatt
    10321 aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatgg
    10381 ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt
    10441 tctggaccga ccggctcggg ttctcccggg acttcgtgga ggacgacttc gccggtgtgg
    10501 tccgggacga cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg ccggacaaca
    10561 ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg
    10621 tgtccacgaa cttccgggac gcctccgggc cggccatgac cgagatcggc gagcagccgt
    10681 gggggcggga gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg
    10741 agcaggactg acacgtgcta cgagatttcg attccaccgc cgccttctat gaaaggttgg
    10801 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
    10861 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
    10921 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
    10981 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
    11041 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
    11101 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
    11161 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
    11221 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
    11281 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
    11341 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
    11401 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
    11461 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
    11521 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
    11581 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
    11641 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
    11701 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
    11761 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
    11821 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
    11881 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
    11941 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
    12001 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
    12061 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
    12121 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
    12181 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
    12241 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
    12301 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
    12361 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
    12421 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
    12481 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
    12541 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
    12601 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
    12661 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
    12721 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
    12781 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
    12841 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
    12901 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
    12961 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
    13021 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
    13081 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
    13141 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
    13201 ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg
    13261 ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga
    13321 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa
    13381 gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg
    13441 ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
    13501 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
    13561 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
    13621 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
    13681 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
    13741 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
//
