LOCUS       sequence-339459-       13124 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  non targeting sgRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13124)
  AUTHORS   Ruf B, Catania VV, Wabitsch S, Ma C, Diggs LP, Zhang Q, Heinrich B, 
            Subramanyam V, Cui LL, Pouzolles M, Evans CN, Chari R, Sakai S, Oh 
            S, Barry CE III, Barber DL, Greten TF
  TITLE     Activating Mucosal-Associated Invariant T Cells Induces a Broad 
            Antitumor Response.
  JOURNAL   Cancer Immunol Res. 2021 Jun 30. pii: 2326-6066.CIR-20-0925. doi: 
            10.1158/2326-6066.CIR-20-0925.
  PUBMED    34193462
REFERENCE   2  (bases 1 to 13124)
  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: "Cancer 
            Immunol Res. 2021 Jun 30. pii: 2326-6066.CIR-20-0925. doi: 
            10.1158/2326-6066.CIR-20-0925."
COMMENT     Sequence Label: pGMC00014
FEATURES             Location/Qualifiers
     source          1..13124
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(52..71)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        243..622
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        624..822
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     777..797
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             840..1020
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1067..1192
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1685..1918
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2103..2147
                     /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    2445..2562
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2613..2861
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2613..2633
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2784..2803
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2883..2958
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        3020..3231
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      3250..3259
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3256..7359
                     /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             3256..7359
                     /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             7360..7407
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7408..7431
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             7441..7497
                     /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             7498..8205
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(7640..7658)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     7908..7927
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    8221..8809
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8274..8294)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8692..8703)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8881..9114
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9140..9157)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9146..9370
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9416..9844
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9503..9522)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9713..9734
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9853..9873)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9858..10187
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10038..10173
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10100..10119
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10235..10282
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10301..10675
                     /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    10805..10926
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10842..10861)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10896..10915
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10975..10991)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10975..10991)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10988..11010)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10999..11015
                     /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(11023..11029,11030..11047,11048..11053))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11068..11089
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11206..11223)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11377..11965)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11457..11476)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12136..12927,12928..12996))
                     /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     12759..12778
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12997..13101)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg
       61 ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga
      121 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa
      181 gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg
      241 ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
      301 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
      361 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
      421 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
      481 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
      541 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
      601 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
      661 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
      721 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
      781 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agcgcgtttt gcctgtactg
      841 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
      901 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
      961 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     1021 gtggcgcccg aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag
     1081 gactcggctt gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc
     1141 aaaaattttg actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa
     1201 gcgggggaga attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa
     1261 atataaatta aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc
     1321 tggcctgtta gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct
     1381 tcagacagga tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt
     1441 gcatcaaagg atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca
     1501 aaacaaaagt aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag
     1561 atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat
     1621 taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg
     1681 gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt
     1741 caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca
     1801 atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca
     1861 agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg
     1921 ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt
     1981 ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag
     2041 aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag
     2101 aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta
     2161 acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag
     2221 gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac
     2281 cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag
     2341 aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac ggatcggcac
     2401 tgcgtgcgcc aattctgcag acaaatggca gtattcatcc acaattttaa aagaaaaggg
     2461 gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa
     2521 actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac
     2581 agcagagatc cagtttggtt aattaaggta ccgagggcct atttcccatg attccttcat
     2641 atttgcatat acgatacaag gctgttagag agataattag aattaatttg actgtaaaca
     2701 caaagatatt agtacaaaat acgtgacgta gaaagtaata atttcttggg tagtttgcag
     2761 ttttaaaatt atgttttaaa atggactatc atatgcttac cgtaacttga aagtatttcg
     2821 atttcttggc tttatatatc ttgtggaaag gacgaaacac cggtgtcgtg atgcgtagac
     2881 gggttttaga gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa
     2941 gtggcaccga gtcggtgctt ttttgaattc gctagctagg tcttgaaagg agtgggaatt
     3001 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
     3061 ggaggggtcg gcaattgatc cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
     3121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
     3181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggaccggttc
     3241 tagagcgctg ccaccatgga caagaagtac agcatcggcc tggacatcgg caccaactct
     3301 gtgggctggg ccgtgatcac cgacgagtac aaggtgccca gcaagaaatt caaggtgctg
     3361 ggcaacaccg accggcacag catcaagaag aacctgatcg gagccctgct gttcgacagc
     3421 ggcgaaacag ccgaggccac ccggctgaag agaaccgcca gaagaagata caccagacgg
     3481 aagaaccgga tctgctatct gcaagagatc ttcagcaacg agatggccaa ggtggacgac
     3541 agcttcttcc acagactgga agagtccttc ctggtggaag aggataagaa gcacgagcgg
     3601 caccccatct tcggcaacat cgtggacgag gtggcctacc acgagaagta ccccaccatc
     3661 taccacctga gaaagaaact ggtggacagc accgacaagg ccgacctgcg gctgatctat
     3721 ctggccctgg cccacatgat caagttccgg ggccacttcc tgatcgaggg cgacctgaac
     3781 cccgacaaca gcgacgtgga caagctgttc atccagctgg tgcagaccta caaccagctg
     3841 ttcgaggaaa accccatcaa cgccagcggc gtggacgcca aggccatcct gtctgccaga
     3901 ctgagcaaga gcagacggct ggaaaatctg atcgcccagc tgcccggcga gaagaagaat
     3961 ggcctgttcg gaaacctgat tgccctgagc ctgggcctga cccccaactt caagagcaac
     4021 ttcgacctgg ccgaggatgc caaactgcag ctgagcaagg acacctacga cgacgacctg
     4081 gacaacctgc tggcccagat cggcgaccag tacgccgacc tgtttctggc cgccaagaac
     4141 ctgtccgacg ccatcctgct gagcgacatc ctgagagtga acaccgagat caccaaggcc
     4201 cccctgagcg cctctatgat caagagatac gacgagcacc accaggacct gaccctgctg
     4261 aaagctctcg tgcggcagca gctgcctgag aagtacaaag agattttctt cgaccagagc
     4321 aagaacggct acgccggcta cattgacggc ggagccagcc aggaagagtt ctacaagttc
     4381 atcaagccca tcctggaaaa gatggacggc accgaggaac tgctcgtgaa gctgaacaga
     4441 gaggacctgc tgcggaagca gcggaccttc gacaacggca gcatccccca ccagatccac
     4501 ctgggagagc tgcacgccat tctgcggcgg caggaagatt tttacccatt cctgaaggac
     4561 aaccgggaaa agatcgagaa gatcctgacc ttccgcatcc cctactacgt gggccctctg
     4621 gccaggggaa acagcagatt cgcctggatg accagaaaga gcgaggaaac catcaccccc
     4681 tggaacttcg aggaagtggt ggacaagggc gcttccgccc agagcttcat cgagcggatg
     4741 accaacttcg ataagaacct gcccaacgag aaggtgctgc ccaagcacag cctgctgtac
     4801 gagtacttca ccgtgtataa cgagctgacc aaagtgaaat acgtgaccga gggaatgaga
     4861 aagcccgcct tcctgagcgg cgagcagaaa aaggccatcg tggacctgct gttcaagacc
     4921 aaccggaaag tgaccgtgaa gcagctgaaa gaggactact tcaagaaaat cgagtgcttc
     4981 gactccgtgg aaatctccgg cgtggaagat cggttcaacg cctccctggg cacataccac
     5041 gatctgctga aaattatcaa ggacaaggac ttcctggaca atgaggaaaa cgaggacatt
     5101 ctggaagata tcgtgctgac cctgacactg tttgaggaca gagagatgat cgaggaacgg
     5161 ctgaaaacct atgcccacct gttcgacgac aaagtgatga agcagctgaa gcggcggaga
     5221 tacaccggct ggggcaggct gagccggaag ctgatcaacg gcatccggga caagcagtcc
     5281 ggcaagacaa tcctggattt cctgaagtcc gacggcttcg ccaacagaaa cttcatgcag
     5341 ctgatccacg acgacagcct gacctttaaa gaggacatcc agaaagccca ggtgtccggc
     5401 cagggcgata gcctgcacga gcacattgcc aatctggccg gcagccccgc cattaagaag
     5461 ggcatcctgc agacagtgaa ggtggtggac gagctcgtga aagtgatggg ccggcacaag
     5521 cccgagaaca tcgtgatcga aatggccaga gagaaccaga ccacccagaa gggacagaag
     5581 aacagccgcg agagaatgaa gcggatcgaa gagggcatca aagagctggg cagccagatc
     5641 ctgaaagaac accccgtgga aaacacccag ctgcagaacg agaagctgta cctgtactac
     5701 ctgcagaatg ggcgggatat gtacgtggac caggaactgg acatcaaccg gctgtccgac
     5761 tacgatgtgg accatatcgt gcctcagagc tttctgaagg acgactccat cgacaacaag
     5821 gtgctgacca gaagcgacaa gaaccggggc aagagcgaca acgtgccctc cgaagaggtc
     5881 gtgaagaaga tgaagaacta ctggcggcag ctgctgaacg ccaagctgat tacccagaga
     5941 aagttcgaca atctgaccaa ggccgagaga ggcggcctga gcgaactgga taaggccggc
     6001 ttcatcaaga gacagctggt ggaaacccgg cagatcacaa agcacgtggc acagatcctg
     6061 gactcccgga tgaacactaa gtacgacgag aatgacaagc tgatccggga agtgaaagtg
     6121 atcaccctga agtccaagct ggtgtccgat ttccggaagg atttccagtt ttacaaagtg
     6181 cgcgagatca acaactacca ccacgcccac gacgcctacc tgaacgccgt cgtgggaacc
     6241 gccctgatca aaaagtaccc taagctggaa agcgagttcg tgtacggcga ctacaaggtg
     6301 tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa tcggcaaggc taccgccaag
     6361 tacttcttct acagcaacat catgaacttt ttcaagaccg agattaccct ggccaacggc
     6421 gagatccgga agcggcctct gatcgagaca aacggcgaaa ccggggagat cgtgtgggat
     6481 aagggccggg attttgccac cgtgcggaaa gtgctgagca tgccccaagt gaatatcgtg
     6541 aaaaagaccg aggtgcagac aggcggcttc agcaaagagt ctatcctgcc caagaggaac
     6601 agcgataagc tgatcgccag aaagaaggac tgggacccta agaagtacgg cggcttcgac
     6661 agccccaccg tggcctattc tgtgctggtg gtggccaaag tggaaaaggg caagtccaag
     6721 aaactgaaga gtgtgaaaga gctgctgggg atcaccatca tggaaagaag cagcttcgag
     6781 aagaatccca tcgactttct ggaagccaag ggctacaaag aagtgaaaaa ggacctgatc
     6841 atcaagctgc ctaagtactc cctgttcgag ctggaaaacg gccggaagag aatgctggcc
     6901 tctgccggcg aactgcagaa gggaaacgaa ctggccctgc cctccaaata tgtgaacttc
     6961 ctgtacctgg ccagccacta tgagaagctg aagggctccc ccgaggataa tgagcagaaa
     7021 cagctgtttg tggaacagca caagcactac ctggacgaga tcatcgagca gatcagcgag
     7081 ttctccaaga gagtgatcct ggccgacgct aatctggaca aagtgctgtc cgcctacaac
     7141 aagcaccggg ataagcccat cagagagcag gccgagaata tcatccacct gtttaccctg
     7201 accaatctgg gagcccctgc cgccttcaag tactttgaca ccaccatcga ccggaagagg
     7261 tacaccagca ccaaagaggt gctggacgcc accctgatcc accagagcat caccggcctg
     7321 tacgagacac ggatcgacct gtctcagctg ggaggcgaca agcgacctgc cgccacaaag
     7381 aaggctggac aggctaagaa gaagaaagat tacaaagacg atgacgataa gggatccggc
     7441 gcaacaaact tctctctgct gaaacaagcc ggagatgtcg aagagaatcc tggaccggtg
     7501 agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac
     7561 atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
     7621 tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc
     7681 tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc
     7741 gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg
     7801 aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag
     7861 ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag
     7921 aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg
     7981 aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc
     8041 aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc
     8101 aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc
     8161 gagggccgcc actccaccgg cggcatggac gagctgtaca agtaaacgcg ttaagtcgac
     8221 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     8281 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     8341 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     8401 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     8461 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     8521 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     8581 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
     8641 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     8701 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     8761 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcg tcgactttaa
     8821 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac
     8881 tggaagggct aattcactcc caacgaagac aagatctgct ttttgcttgt actgggtctc
     8941 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     9001 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     9061 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagggccc
     9121 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     9181 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     9241 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     9301 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     9361 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     9421 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     9481 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     9541 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     9601 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     9661 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     9721 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     9781 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     9841 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     9901 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     9961 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
    10021 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
    10081 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
    10141 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
    10201 cttgtatatc cattttcgga tctgatcagc acgtgttgac aattaatcat cggcatagta
    10261 tatcggcata gtataatacg acaaggtgag gaactaaacc atggccaagt tgaccagtgc
    10321 cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc gagttctgga ccgaccggct
    10381 cgggttctcc cgggacttcg tggaggacga cttcgccggt gtggtccggg acgacgtgac
    10441 cctgttcatc agcgcggtcc aggaccaggt ggtgccggac aacaccctgg cctgggtgtg
    10501 ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg
    10561 ggacgcctcc gggccggcca tgaccgagat cggcgagcag ccgtgggggc gggagttcgc
    10621 cctgcgcgac ccggccggca actgcgtgca cttcgtggcc gaggagcagg actgacacgt
    10681 gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
    10741 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
    10801 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
    10861 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
    10921 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
    10981 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
    11041 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
    11101 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
    11161 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
    11221 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
    11281 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
    11341 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
    11401 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
    11461 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
    11521 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
    11581 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
    11641 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
    11701 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
    11761 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
    11821 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
    11881 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
    11941 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
    12001 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
    12061 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
    12121 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
    12181 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
    12241 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
    12301 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
    12361 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
    12421 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
    12481 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
    12541 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
    12601 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
    12661 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
    12721 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
    12781 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
    12841 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
    12901 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
    12961 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
    13021 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
    13081 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccac
//
