LOCUS       sequence-272060-        9750 bp    DNA     circular SYN 14-APR-2025
DEFINITION  CMV promoter expression plasmid for 
            TadA*(V82G)-nCas9-pmCDA1(R187W)-P2A-EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9750)
  AUTHORS   Grunewald J, Zhou R, Lareau CA, Garcia SP, Iyer S, Miller BR, 
            Langner LM, Hsu JY, Aryee MJ, Joung JK
  TITLE     A dual-deaminase CRISPR base editor enables concurrent adenine and 
            cytosine editing.
  JOURNAL   Nat Biotechnol. 2020 Jun 1. pii: 10.1038/s41587-020-0535-y. doi: 
            10.1038/s41587-020-0535-y.
  PUBMED    32483364
REFERENCE   2  (bases 1 to 9750)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Biotechnol. 2020 Jun 1. pii: 10.1038/s41587-020-0535-y. doi: 
            10.1038/s41587-020-0535-y."
COMMENT     Sequence Label: SPACE-Delta-UGI (pRZ1836)
FEATURES             Location/Qualifiers
     source          1..9750
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     220..239
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     473..490
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    607..628
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(643..648,649..666,667..673)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    681..697
                     /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     686..708
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     705..721
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     705..721
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    792..1016
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1005..1022
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(1045..1062)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(1071..1091)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(join(1104..1814,1815..1817,1818..1820))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(1104..1814,1815..1817,1818..1820))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1146..1167)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1495..1514
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             complement(1821..1877)
                     /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             complement(2604..2624)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(join(2667..6740,6741..6743,6744..6767))
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(2667..6767)
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(7362..7382)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(7431..7450)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7432..7450)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(7492..7695)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(7521..7541)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        7696..8075
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     8247..8266
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        8345..8449
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8450..8518,8519..9310)
                     /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(8668..8687)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(9481..9750,1..319)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
       61 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
      121 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
      181 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
      241 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
      301 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
      361 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
      421 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
      481 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
      541 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
      601 cgcaattaat gtgagttagc tcactcatta ggcaccctag gctttacact ttatgcttcc
      661 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
      721 ccatgattac gccaagctct agctagaggt cgacggtatc gagccccagc tggttctttc
      781 cgcctcagaa gccatagagc ccaccgcatc cccagcatgc ctgctattgt cttcccaatc
      841 ctcccccttg ctgtcctgcc ccaccccacc ccccagaata gaatgacacc tactcagaca
      901 atgcgatgca atttcctcat tttattagga aaggacagtg ggagtggcac cttccagggt
      961 caaggaaggc acgggggagg ggcaaacaac agatggctgg caactagaag gcacagtcga
     1021 ggctgatcag cgggtttaaa ctcaatggtg atggtgatga tgaccggtta gactttcctc
     1081 ttcttcttgg gagaaccacc agacttgtac agctcgtcca tgccgagagt gatcccggcg
     1141 gcggtcacga actccagcag gaccatgtga tcgcgcttct cgttggggtc tttgctcagg
     1201 gcggactggg tgctcaggta gtggttgtcg ggcagcagca cggggccgtc gccgatgggg
     1261 gtgttctgct ggtagtggtc ggcgagctgc acgctgccgt cctcgatgtt gtggcggatc
     1321 ttgaagttca ccttgatgcc gttcttctgc ttgtcggcca tgatatagac gttgtggctg
     1381 ttgtagttgt actccagctt gtgccccagg atgttgccgt cctccttgaa gtcgatgccc
     1441 ttcagctcga tgcggttcac cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg
     1501 tagttgccgt cgtccttgaa gaagatggtg cgctcctgga cgtagccttc gggcatggcg
     1561 gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc ggctgaagca ctgcactcca
     1621 taggtcaggg tggtcacgag ggtgggccag ggcacgggca gcttgccggt ggtgcagatg
     1681 aacttcaggg tcagcttgcc gtaggtggca tcgccctcgc cctcgccgga cacgctgaac
     1741 ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg gcaccacccc ggtgaacagc
     1801 tcctcgccct tgctcaccat aggtccaggg ttctcctcca cgtctccagc ctgcttcagc
     1861 aggctgaagt tagtagctcc gcttccaacg gctggagact tagtggtatg cagaatcttg
     1921 acctggatca tgatgctcag ctcggatctc cacttctccg cacgtttcag tgttttctcc
     1981 aaccagcggt tttcattgag ctggttatga gagctttgga tgaaaatctt ccgacaacac
     2041 tgatagtgct cggaaaccat cacgttcagg cctacaccat tgtccctcag attccagagt
     2101 cctatctggt tccgggcatt cttctcgtag tacagcttgc aggcccagat tttcaatgtg
     2161 tgcccgtttc cccgaagttc ctgattatac cattccagaa ttttctcggc acaatcagcg
     2221 caaggactcc agctggaata ccagttgatc gtaaactgcc caggattatc ccgaagatac
     2281 tcttccacct ttcggatact gaatatctca gcgtgtattc ctctctcggt gccactttgg
     2341 ggcttgttga cggcatagcc ccaaaagcaa gcccttcttt cgccccttcg cttcaactca
     2401 aacagcacgt agcagcggtg tgacacagat ttcttgttgt tgaagaactg cttcttgaag
     2461 gtgtaaatat ccagtttctc atgaatgcgc acgtactcgg cgtctgtgga tcctccggag
     2521 gagccgccgg agctctctgg tgtggcggac tcgcttgtgc ctggtgtctc gcttccagag
     2581 gatcctccgc tagatcctcc agagactttc ctcttcttct tgggctcgaa ttcgctgccg
     2641 tcggcggttc tttttgagcc gccagagtca cctcccagct gagacaggtc gatccgtgtc
     2701 tcgtacaggc cggtgatgct ctggtggatc agggtggcgt ccagcacctc tttggtgctg
     2761 gtgtacctct tccggtcgat ggtggtgtca aagtacttga aggcggcagg ggctcccaga
     2821 ttggtcaggg taaacaggtg gatgatattc tcggcctgct ctctgatggg cttatcccgg
     2881 tgcttgttgt aggcggacag cactttgtcc agattagcgt cggccaggat cactctcttg
     2941 gagaactcgc tgatctgctc gatgatctcg tccaggtagt gcttgtgctg ttccacaaac
     3001 agctgtttct gctcattatc ctcgggggag cccttcagct tctcatagtg gctggccagg
     3061 tacaggaagt tcacatattt ggagggcagg gccagttcgt ttcccttctg cagttcgccg
     3121 gcagaggcca gcattctctt ccggccgttt tccagctcga acagggagta cttaggcagc
     3181 ttgatgatca ggtccttttt cacttctttg tagcccttgg cttccagaaa gtcgatggga
     3241 ttcttctcga agctgcttct ttccatgatg gtgatcccca gcagctcttt cacactcttc
     3301 agtttcttgg acttgccctt ttccactttg gccaccacca gcacagaata ggccacggtg
     3361 gggctgtcga agccgccgta cttcttaggg tcccagtcct tctttctggc gatcagctta
     3421 tcgctgttcc tcttgggcag gatagactct ttgctgaagc cgcctgtctg cacctcggtc
     3481 tttttcacga tattcacttg gggcatgctc agcactttcc gcacggtggc aaaatcccgg
     3541 cccttatccc acacgatctc cccggtttcg ccgtttgtct cgatcagagg ccgcttccgg
     3601 atctcgccgt tggccagggt aatctcggtc ttgaaaaagt tcatgatgtt gctgtagaag
     3661 aagtacttgg cggtagcctt gccgatttcc tgctcgctct tggcgatcat cttccgcacg
     3721 tcgtacacct tgtagtcgcc gtacacgaac tcgctttcca gcttagggta ctttttgatc
     3781 agggcggttc ccacgacggc gttcaggtag gcgtcgtggg cgtggtggta gttgttgatc
     3841 tcgcgcactt tgtaaaactg gaaatccttc cggaaatcgg acaccagctt ggacttcagg
     3901 gtgatcactt tcacttcccg gatcagcttg tcattctcgt cgtacttagt gttcatccgg
     3961 gagtccagga tctgtgccac gtgctttgtg atctgccggg tttccaccag ctgtctcttg
     4021 atgaagccgg ccttatccag ttcgctcagg ccgcctctct cggccttggt cagattgtcg
     4081 aactttctct gggtaatcag cttggcgttc agcagctgcc gccagtagtt cttcatcttc
     4141 ttcacgacct cttcggaggg cacgttgtcg ctcttgcccc ggttcttgtc gcttctggtc
     4201 agcaccttgt tgtcgatgga gtcgtccttc agaaagctct gaggcacgat atggtccaca
     4261 tcgtagtcgg acagccggtt gatgtccagt tcctggtcca cgtacatatc ccgcccattc
     4321 tgcaggtagt acaggtacag cttctcgttc tgcagctggg tgttttccac ggggtgttct
     4381 ttcaggatct ggctgcccag ctctttgatg ccctcttcga tccgcttcat tctctcgcgg
     4441 ctgttcttct gtcccttctg ggtggtctgg ttctctctgg ccatttcgat cacgatgttc
     4501 tcgggcttgt gccggcccat cactttcacg agctcgtcca ccaccttcac tgtctgcagg
     4561 atgcccttct taatggcggg gctgccggcc agattggcaa tgtgctcgtg caggctatcg
     4621 ccctggccgg acacctgggc tttctggatg tcctctttaa aggtcaggct gtcgtcgtgg
     4681 atcagctgca tgaagtttct gttggcgaag ccgtcggact tcaggaaatc caggattgtc
     4741 ttgccggact gcttgtcccg gatgccgttg atcagcttcc ggctcagcct gccccagccg
     4801 gtgtatctcc gccgcttcag ctgcttcatc actttgtcgt cgaacaggtg ggcataggtt
     4861 ttcagccgtt cctcgatcat ctctctgtcc tcaaacagtg tcagggtcag cacgatatct
     4921 tccagaatgt cctcgttttc ctcattgtcc aggaagtcct tgtccttgat aattttcagc
     4981 agatcgtggt atgtgcccag ggaggcgttg aaccgatctt ccacgccgga gatttccacg
     5041 gagtcgaagc actcgatttt cttgaagtag tcctctttca gctgcttcac ggtcactttc
     5101 cggttggtct tgaacagcag gtccacgatg gcctttttct gctcgccgct caggaaggcg
     5161 ggctttctca ttccctcggt cacgtatttc actttggtca gctcgttata cacggtgaag
     5221 tactcgtaca gcaggctgtg cttgggcagc accttctcgt tgggcaggtt cttatcgaag
     5281 ttggtcatcc gctcgatgaa gctctgggcg gaagcgccct tgtccaccac ttcctcgaag
     5341 ttccaggggg tgatggtttc ctcgctcttt ctggtcatcc aggcgaatct gctgtttccc
     5401 ctggccagag ggcccacgta gtaggggatg cggaaggtca ggatcttctc gatcttttcc
     5461 cggttgtcct tcaggaatgg gtaaaaatct tcctgccgcc gcagaatggc gtgcagctct
     5521 cccaggtgga tctggtgggg gatgctgccg ttgtcgaagg tccgctgctt ccgcagcagg
     5581 tcctctctgt tcagcttcac gagcagttcc tcggtgccgt ccatcttttc caggatgggc
     5641 ttgatgaact tgtagaactc ttcctggctg gctccgccgt caatgtagcc ggcgtagccg
     5701 ttcttgctct ggtcgaagaa aatctctttg tacttctcag gcagctgctg ccgcacgaga
     5761 gctttcagca gggtcaggtc ctggtggtgc tcgtcgtatc tcttgatcat agaggcgctc
     5821 aggggggcct tggtgatctc ggtgttcact ctcaggatgt cgctcagcag gatggcgtcg
     5881 gacaggttct tggcggccag aaacaggtcg gcgtactggt cgccgatctg ggccagcagg
     5941 ttgtccaggt cgtcgtcgta ggtgtccttg ctcagctgca gtttggcatc ctcggccagg
     6001 tcgaagttgc tcttgaagtt gggggtcagg cccaggctca gggcaatcag gtttccgaac
     6061 aggccattct tcttctcgcc gggcagctgg gcgatcagat tttccagccg tctgctcttg
     6121 ctcagtctgg cagacaggat ggccttggcg tccacgccgc tggcgttgat ggggttttcc
     6181 tcgaacagct ggttgtaggt ctgcaccagc tggatgaaca gcttgtccac gtcgctgttg
     6241 tcggggttca ggtcgccctc gatcaggaag tggccccgga acttgatcat gtgggccagg
     6301 gccagataga tcagccgcag gtcggccttg tcggtgctgt ccaccagttt ctttctcagg
     6361 tggtagatgg tggggtactt ctcgtggtag gccacctcgt ccacgatgtt gccgaagatg
     6421 gggtgccgct cgtgcttctt atcctcttcc accaggaagg actcttccag tctgtggaag
     6481 aagctgtcgt ccaccttggc catctcgttg ctgaagatct cttgcagata gcagatccgg
     6541 ttcttccgtc tggtgtatct tcttctggcg gttctcttca gccgggtggc ctcggctgtt
     6601 tcgccgctgt cgaacagcag ggctccgatc aggttcttct tgatgctgtg ccggtcggtg
     6661 ttgcccagca ccttgaattt cttgctgggc accttgtact cgtcggtgat cacggcccag
     6721 cccacagagt tggtgccgat ggccaggccg atgctgtact tcttgtctga ccccccgctg
     6781 ctgcccccgc tgctttcagg tgttgcgctc tcgcttgtgc caggtgtctc agagccagag
     6841 gagcctccgc tagatcctcc ggagtcggtg gagctctggg ccttcttctg agcattgaac
     6901 acctgtctag gcatccgaaa gaaatagcac agcagggcgg cacattcatc tgccaggatt
     6961 ccctcggtaa tttcgacgcg gtgattcatg ccggggtagt gcagcacgtc catcagggag
     7021 cctgcggcgc cggtttttgc gttcctcacg ccaaacacca cgcggccgat cctagagtgg
     7081 atcatggcgc cggcgcacat cacgcaaggc tcgaatgtgc cgtacagggt ggcgtcaatc
     7141 agtctgtagt tctgcatgac caggccgccc tgtctcaggg ccataatttc ggcatgggct
     7201 gttgggtcgt gcaggccgat ggctctgttc cagccctcgc cgatcactct attgttcagc
     7261 accagcacgg ctcccacagg cacctccctc tcatcgcgtg ccctcttggc cagggtcagg
     7321 gcatgtctca tccagtactc gtgggaaaac tccacctcag agactttccg cttcttcttt
     7381 ggtgactcga actcgcttcc gtcggctgtc cgtttcatgg tggcggctct ccctatagtg
     7441 agtcgtatta gcggccgcgg atctctagcg gatctgacgg ttcactaaac cagctctgct
     7501 tatatagacc tcccaccgta cacgcctacc gcccatttgc gtcaatgggg cggagttgtt
     7561 acgacatttt ggaaagtccc gttgattttg gtgccaaaac aaactcccat tgacgtcaat
     7621 ggggtggaga cttggaaatc cccgtgagtc aaaccgctat ccacgcccat tgatgtactg
     7681 ccaaaaccgc atcaccatgg taatagcgat gactaatacg tagatgtact gccaagtagg
     7741 aaagtcccat aaggtcatgt actgggcata atgccaggcg ggccatttac cgtcattgac
     7801 gtcaataggg ggcgtacttg gcatatgata cacttgatgt actgccaagt gggcagttta
     7861 ccgtaaatac tccacccatt gacgtcaatg gaaagtccct attggcgtta ctatgggaac
     7921 atacgtcatt attgacgtca atgggcgggg gtcgttgggc ggtcagccag gcgggccatt
     7981 taccgtaagt tatgtaacgc ggaactccat atatgggcta tgaactaatg accccgtaat
     8041 tgattactat taataactag tcaataatca atgtcaacgc gtatatctgg cccgtacatc
     8101 gcgaagcagc gcaaaacgcc taaccctaag cagattcttc atgcaattgt cggtcaagcc
     8161 ttgccttgtt gtagcttaaa ttttgctcgc gcactactca gcgacctcca acacacaagc
     8221 agggagcaga tactggctta actatgcggc atcagagcag attgtactga gagtcgaccc
     8281 taggggatcg ggagatcgat ctcccgatcc gtcgacgtca ggtggcactt ttcggggaaa
     8341 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     8401 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     8461 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     8521 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     8581 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     8641 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     8701 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     8761 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     8821 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     8881 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     8941 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     9001 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     9061 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     9121 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     9181 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     9241 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     9301 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     9361 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     9421 ttaacgtgag ttttcgttcc actgagtgtc agaccccgta gaaaagatca aaggatcttc
     9481 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     9541 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     9601 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     9661 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     9721 tgccagtggc gataagtcgt gtcttaccgg
//
