LOCUS       Exported               13171 bp ds-DNA     circular SYN 26-JUN-2025
DEFINITION  Lentiviral vector for constitutive expression of NG-ABE8e-P2A-GFP in
            mammalian cells (codon optimized).
ACCESSION   .
VERSION     .
KEYWORDS    pLenti NG-ABE8e_P2A_EGFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13171)
  AUTHORS   Wang P, Sen R, Buchholz F, Sayed S
  TITLE     A base editing platform for the correction of cancer driver 
  JOURNAL   Genome Biology, 2025
REFERENCE   2  (bases 1 to 13171)
  AUTHORS   Zafra MP, Schatoff EM, Katti A, Foronda M, Breinig M, Schweitzer AY,
            Simon A, Han T, Goswami S, Montgomery E, Thibado J, Kastenhuber ER, 
            Sanchez-Rivera FJ, Shi J, Vakoc CR, Lowe SW, Tschaharganeh DF, Dow 
            LE
  TITLE     Optimized base editors enable efficient editing in cells, organoids 
            and mice.
  JOURNAL   Nat Biotechnol. 2018 Jul 3. pii: nbt.4194. doi: 10.1038/nbt.4194.
  PUBMED    29969439
REFERENCE   3  (bases 1 to 13171)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Jun 26, 2025 from SnapGene 5.1.7
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Genome 
            Biology, 2025"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Nat 
            Biotechnol. 2018 Jul 3. pii: nbt.4194. doi: 10.1038/nbt.4194."
FEATURES             Location/Qualifiers
     source          1..13171
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /note="color: #ffffff"
     promoter        1..227
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
                     /note="color: #ffffff; direction: RIGHT"
     LTR             228..408
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /note="color: #ffe4c4"
     misc_feature    455..580
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virustype
                     1"
                     /note="color: #ffe4c4"
     misc_feature    1073..1306
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
                     /note="color: #ffe4c4"
     misc_feature    1719..1836
                     /label=cPPT/CTS
                     /note="central polypurine tract and central 
                     terminationsequence of HIV-1"
                     /note="color: #ffe4c4"
     promoter        1905..2116
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factorEF-1-alpha"
                     /note="color: #ffffff; direction: RIGHT"
     misc_feature    2131..7090
                     /label=NGCas9.ABE8e
                     /note="color: #ffffff; direction: RIGHT"
     primer_bind     2142..2162
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
                     /note="color: black; sequence: CGCAAATGGGCGGTAGGCGTG"
     primer_bind     2233..2252
                     /label=T7
                     /note="T7 promoter, forward primer"
                     /note="color: black; sequence: TAATACGACTCACTATAGGG"
     promoter        2233..2251
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     misc_feature    2256..2268
                     /label=Kozak
                     /note="color: #a6acb3"
     CDS             2268..2321
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian 
                     virus40) large T antigen"
                     /label=bipartite SV40 NLS
                     /note="color: #c0c0c0"
                     /translation="KRTADGSEFESPKKKRKV"
     CDS             2322..2822
                     /codon_start=1
                     /label=Deaminase
                     /note="color: #993366"
                     /translation="SEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWN
                     RAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVFG
                     VRNSKRGAAGSLMNVLNYPGMNHRVEITEGILADECAALLCDFYRMPRQVFNAQKKAQS
                     SINS"
     CDS             2823..2915
                     /codon_start=1
                     /label=Linker
                     /note="color: #c0c0c0"
                     /translation="GGSSGGSSGSETPGTSESATPESSGGSSGGS"
     CDS             2916..7016
                     /codon_start=1
                     /label=NG-Cas9
                     /note="color: #993366"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SIRPKRNSDKLIARKKDWDPKKYGGFVSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASARFLQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPRAFKYFDTTIDRKVYRSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     misc_feature    2941
                     /label=D10A
                     /note="D10A mutation, inactivates RuvC; nuclease 
                     -->nickase"
                     /note="color: #ff0000"
     primer_bind     complement(6519..6539)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
                     /note="color: black; sequence: CGTCGCCGTCCAGCTCGACCAG"
     CDS             7029..7079
                     /codon_start=1
                     /label=bipartitie SV40 NLS
                     /note="color: #993366"
                     /translation="KRTADGSEFEPKKKRKV"
     CDS             7101..7157
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /note="color: #cc99b2
                     Cleavage site after base 7154"
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             7158..7877
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /note="This feature has 3 segments:
                        1: 7158 .. 7160 / #05fd14
                        2: 7161 .. 7163 / #05fd14 / 1a
                        3: 7164 .. 7877 / #05fd14"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7203..7224)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
                     /note="color: black; sequence: CGTCGCCGTCCAGCTCGACCAG"
     primer_bind     complement(7464..7483)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
                     /note="color: black; sequence: GTCTTGTAGTTGCCGTCGTC"
     primer_bind     7811..7832
                     /label=EGFP-C
                     /note="EGFP, forward primer"
                     /note="color: black; sequence: CATGGTCCTGCTGGAGTTCGTG"
     promoter        7889..8388
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /note="color: #ffffff; direction: RIGHT"
     primer_bind     complement(7909..7929)
                     /label=Primer 13 for PB
                     /note="color: black; sequence: gcatgctccagactgccttgg; 
                     added: 2024-10-16"
     primer_bind     complement(7924..7947)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
                     /note="color: black; sequence: CAGCGGGGCTGCTAAAGCGCATGC"
     primer_bind     complement(7924..7942)
                     /label=Primer 15 for PB
                     /note="color: black; sequence: gggctgctaaagcgcatgc; added: 
                     2024-10-16"
     primer_bind     complement(7949..7970)
                     /label=Primer 14 for PB
                     /note="color: black; sequence: ccacttgtgtagcgccaagtgc; 
                     added: 2024-10-16"
     primer_bind     complement(7972..7992)
                     /label=Primer 16 for PB
                     /note="color: black; sequence: ggaatgtgtgcgaggccagag; 
                     added: 2024-10-16"
     primer_bind     8321..8340
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
                     /note="color: black; sequence: CATTCTGCACGCTTCAAAAG"
     CDS             8409..9008
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /note="color: #ccffcc"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(8409..8428)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. 
                     Alsocalled puro-variant-R"
                     /note="color: black; sequence: GTGGGCTTGTACTCGGTCAT"
     primer_bind     8905..8925
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
                     /note="color: black; sequence: GCAACCTCCCCTTCTACGAGC"
     misc_feature    9024..9612
                     /label=WPRE
                     /note="woodchuck hepatitis virus 
                     posttranscriptionalregulatory element"
                     /note="color: #ffffff"
     primer_bind     complement(9077..9097)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
                     /note="color: black; sequence: CATAGCGTAAAAGGAGCAACA"
     CDS             complement(9495..9506)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /note="color: #cc99b2
                     Cleavage site after base 9494"
                     /translation="IEGR"
     LTR             9684..9917
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
                     /note="color: #ffe4c4"
     polyA_signal    9995..10116
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     complement(10032..10051)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
                     /note="color: black; sequence: GAAATTTGTGATGCTATTGC"
     primer_bind     10086..10105
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
                     /note="color: black; sequence: GTGGTTTGTCCAAACTCATC"
     rep_origin      10156..10291
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     primer_bind     10218..10237
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
                     /note="color: black; sequence: TATTTATGCAGAGGCCGAGG"
     primer_bind     complement(10241..10252)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
                     /note="color: black; sequence: TATTTATGCAGAGGCCGAGG"
     primer_bind     complement(10311..10330)
                     /label=T7
                     /note="T7 promoter, forward primer"
                     /note="color: black; sequence: TAATACGACTCACTATAGGG"
     promoter        complement(10312..10330)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: LEFT"
     primer_bind     complement(10340..10357)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 
                     (-21)Forward"
                     /note="color: black; sequence: TGTAAAACGACGGCCAGT"
     primer_bind     complement(10340..10356)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     primer_bind     complement(10349..10371)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
                     /note="color: black; sequence: CCCAGTCACGACGTTGTAAAACG"
     rep_origin      10498..10953
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; 
                     arrowindicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     primer_bind     complement(10585..10604)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
                     /note="color: black; sequence: AGGGAAGAAAGCGAAAGGAG"
     primer_bind     10795..10816
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
                     /note="color: black; sequence: GTGGACTCTTGTTCCAAACTGG"
     promoter        10979..11083
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             11084..11944
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 11084 .. 11152 / #ccffcc / signal sequence
                        2: 11153 .. 11944 / #ccffcc
                     Cleavage site after base 11152"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(11302..11321)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
                     /note="color: black; sequence: ATAATACCGCGCCACATAGC"
     rep_origin      12115..12703
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     primer_bind     12604..12623
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
                     /note="color: black; sequence: GGGAAACGCCTGGTATCTTT"
     primer_bind     12857..12874
                     /label=L4440
                     /note="L4440 vector, forward primer"
                     /note="color: black; sequence: AGCGAGTCAGTGAGCGAG"
     protein_bind    12991..13012
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the 
                     presenceof cAMP."
                     /note="color: #31849b"
     promoter        13027..13057
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This forward directional feature has 3 segments:
                        1: 13027 .. 13032 / #ffffff / -35
                        2: 13033 .. 13050 / #ffffff
                        3: 13051 .. 13057 / #ffffff / -10"
     protein_bind    13065..13081
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator 
                     toinhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /note="color: #31849b"
     primer_bind     13070..13092
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
                     /note="color: black; sequence: AGCGGATAACAATTTCACACAGG"
     primer_bind     13089..13105
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: RIGHT"
     primer_bind     13089..13105
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
                     /note="color: black; sequence: CAGGAAACAGCTATGAC"
     primer_bind     13124..13144
                     /label=T3
                     /note="T3 promoter, forward primer"
                     /note="color: black; sequence: GCAATTAACCCTCACTAAAGG"
     promoter        13126..13144
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
ORIGIN
        1 tgtagtctta tgcaatactc ttgtagtctt gcaacatggt aacgatgagt tagcaacatg
       61 ccttacaagg agagaaaaag caccgtgcat gccgattggt ggaagtaagg tggtacgatc
      121 gtgccttatt aggaaggcaa cagacgggtc tgacatggat tggacgaacc actgaattgc
      181 cgcattgcag agatattgta tttaagtgcc tagctcgata cataaacggg tctctctggt
      241 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
      301 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
      361 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa
      421 cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc
      481 tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac
      541 tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat
      601 tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa
      661 acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga
      721 aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc
      781 agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat
      841 agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa
      901 gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca
      961 attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac
     1021 ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt
     1081 tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga
     1141 cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg
     1201 ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg
     1261 caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt
     1321 gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat
     1381 ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt
     1441 acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac
     1501 aagaattatt ggaattaaat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt
     1561 ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag
     1621 tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc
     1681 agacccacct cccaaccccg aggggaccca gaccacaatt ttaaaagaaa aggggggatt
     1741 ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa
     1801 gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag ggacagcaga
     1861 gatccacttt ggcgccggct cgagtggctc cggtgcccgt cagtgggcag agcgcacatc
     1921 gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaaccggtg cctagagaag
     1981 gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg
     2041 tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt
     2101 tgccgccaga acacaggtgt cgtgacgcgg gatccggtac ccgcaaatgg gcggtaggcg
     2161 tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagag
     2221 atccgcggcc gctaatacga ctcactatag ggagagccgc caccatgaaa cggacagccg
     2281 acggaagcga gttcgagtca ccaaagaaga agcggaaagt ctctgaggtg gagttttccc
     2341 acgagtactg gatgagacat gccctgaccc tggccaagag ggcacgggat gagagggagg
     2401 tgcctgtggg agccgtgctg gtgctgaaca atagagtgat cggcgagggc tggaacagag
     2461 ccatcggcct gcacgaccca acagcccatg ccgaaattat ggccctgaga cagggcggcc
     2521 tggtcatgca gaactacaga ctgattgacg ccaccctgta cgtgacattc gagccttgcg
     2581 tgatgtgcgc cggcgccatg atccactcta ggatcggccg cgtggtgttt ggcgtgagga
     2641 actcaaaaag aggcgccgca ggctccctga tgaacgtgct gaactacccc ggcatgaatc
     2701 accgcgtcga aattaccgag ggaatcctgg cagatgaatg tgccgccctg ctgtgcgatt
     2761 tctatcggat gcctagacag gtgttcaatg ctcagaagaa ggcccagagc tccatcaact
     2821 ccggaggatc tagcggaggc tcctctggct ctgagacacc tggcacaagc gagagcgcaa
     2881 cacctgaaag cagcgggggc agcagcgggg ggtcagacaa gaagtacagc atcggcctgg
     2941 ccatcggcac caactctgtg ggctgggccg tgatcaccga cgagtacaag gtgcccagca
     3001 agaaattcaa ggtgctgggc aacaccgacc ggcacagcat caagaagaac ctgatcggag
     3061 ccctgctgtt cgacagcggc gaaacagccg aggccacccg gctgaagaga accgccagaa
     3121 gaagatacac cagacggaag aaccggatct gctatctgca agagatcttc agcaacgaga
     3181 tggccaaggt ggacgacagc ttcttccaca gactggaaga gtccttcctg gtggaagagg
     3241 ataagaagca cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg
     3301 agaagtaccc caccatctac cacctgagaa agaaactggt ggacagcacc gacaaggccg
     3361 acctgcggct gatctatctg gccctggccc acatgatcaa gttccggggc cacttcctga
     3421 tcgagggcga cctgaacccc gacaacagcg acgtggacaa gctgttcatc cagctggtgc
     3481 agacctacaa ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg
     3541 ccatcctgtc tgccagactg agcaagagca gacggctgga aaatctgatc gcccagctgc
     3601 ccggcgagaa gaagaatggc ctgttcggaa acctgattgc cctgagcctg ggcctgaccc
     3661 ccaacttcaa gagcaacttc gacctggccg aggatgccaa actgcagctg agcaaggaca
     3721 cctacgacga cgacctggac aacctgctgg cccagatcgg cgaccagtac gccgacctgt
     3781 ttctggccgc caagaacctg tccgacgcca tcctgctgag cgacatcctg agagtgaaca
     3841 ccgagatcac caaggccccc ctgagcgcct ctatgatcaa gagatacgac gagcaccacc
     3901 aggacctgac cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag tacaaagaga
     3961 ttttcttcga ccagagcaag aacggctacg ccggctacat tgacggcgga gccagccagg
     4021 aagagttcta caagttcatc aagcccatcc tggaaaagat ggacggcacc gaggaactgc
     4081 tcgtgaagct gaacagagag gacctgctgc ggaagcagcg gaccttcgac aacggcagca
     4141 tcccccacca gatccacctg ggagagctgc acgccattct gcggcggcag gaagattttt
     4201 acccattcct gaaggacaac cgggaaaaga tcgagaagat cctgaccttc cgcatcccct
     4261 actacgtggg ccctctggcc aggggaaaca gcagattcgc ctggatgacc agaaagagcg
     4321 aggaaaccat caccccctgg aacttcgagg aagtggtgga caagggcgct tccgcccaga
     4381 gcttcatcga gcggatgacc aacttcgata agaacctgcc caacgagaag gtgctgccca
     4441 agcacagcct gctgtacgag tacttcaccg tgtataacga gctgaccaaa gtgaaatacg
     4501 tgaccgaggg aatgagaaag cccgccttcc tgagcggcga gcagaaaaag gccatcgtgg
     4561 acctgctgtt caagaccaac cggaaagtga ccgtgaagca gctgaaagag gactacttca
     4621 agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct
     4681 ccctgggcac ataccacgat ctgctgaaaa ttatcaagga caaggacttc ctggacaatg
     4741 aggaaaacga ggacattctg gaagatatcg tgctgaccct gacactgttt gaggacagag
     4801 agatgatcga ggaacggctg aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc
     4861 agctgaagcg gcggagatac accggctggg gcaggctgag ccggaagctg atcaacggca
     4921 tccgggacaa gcagtccggc aagacaatcc tggatttcct gaagtccgac ggcttcgcca
     4981 acagaaactt catgcagctg atccacgacg acagcctgac ctttaaagag gacatccaga
     5041 aagcccaggt gtccggccag ggcgatagcc tgcacgagca cattgccaat ctggccggca
     5101 gccccgccat taagaagggc atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag
     5161 tgatgggccg gcacaagccc gagaacatcg tgatcgaaat ggccagagag aaccagacca
     5221 cccagaaggg acagaagaac agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag
     5281 agctgggcag ccagatcctg aaagaacacc ccgtggaaaa cacccagctg cagaacgaga
     5341 agctgtacct gtactacctg cagaatgggc gggatatgta cgtggaccag gaactggaca
     5401 tcaaccggct gtccgactac gatgtggacc atatcgtgcc tcagagcttt ctgaaggacg
     5461 actccatcga caacaaggtg ctgaccagaa gcgacaagaa ccggggcaag agcgacaacg
     5521 tgccctccga agaggtcgtg aagaagatga agaactactg gcggcagctg ctgaacgcca
     5581 agctgattac ccagagaaag ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg
     5641 aactggataa ggccggcttc atcaagagac agctggtgga aacccggcag atcacaaagc
     5701 acgtggcaca gatcctggac tcccggatga acactaagta cgacgagaat gacaagctga
     5761 tccgggaagt gaaagtgatc accctgaagt ccaagctggt gtccgatttc cggaaggatt
     5821 tccagtttta caaagtgcgc gagatcaaca actaccacca cgcccacgac gcctacctga
     5881 acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt
     5941 acggcgacta caaggtgtac gacgtgcgga agatgatcgc caagagcgag caggaaatcg
     6001 gcaaggctac cgccaagtac ttcttctaca gcaacatcat gaactttttc aagaccgaga
     6061 ttaccctggc caacggcgag atccggaagc ggcctctgat cgagacaaac ggcgaaaccg
     6121 gggagatcgt gtgggataag ggccgggatt ttgccaccgt gcggaaagtg ctgagcatgc
     6181 cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc aaagagtcta
     6241 tcaggcccaa gaggaacagc gataagctga tcgccagaaa gaaggactgg gaccctaaga
     6301 agtacggcgg cttcgtcagc cccaccgtgg cctattctgt gctggtggtg gccaaagtgg
     6361 aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct gctggggatc accatcatgg
     6421 aaagaagcag cttcgagaag aatcccatcg actttctgga agccaagggc tacaaagaag
     6481 tgaaaaagga cctgatcatc aagctgccta agtactccct gttcgagctg gaaaacggcc
     6541 ggaagagaat gctggcctct gccagattcc tgcagaaggg aaacgaactg gccctgccct
     6601 ccaaatatgt gaacttcctg tacctggcca gccactatga gaagctgaag ggctcccccg
     6661 aggataatga gcagaaacag ctgtttgtgg aacagcacaa gcactacctg gacgagatca
     6721 tcgagcagat cagcgagttc tccaagagag tgatcctggc cgacgctaat ctggacaaag
     6781 tgctgtccgc ctacaacaag caccgggata agcccatcag agagcaggcc gagaatatca
     6841 tccacctgtt taccctgacc aatctgggag cccctagggc cttcaagtac tttgacacca
     6901 ccatcgaccg gaaggtgtac aggagcacca aagaggtgct ggacgccacc ctgatccacc
     6961 agagcatcac cggcctgtac gagacacgga tcgacctgtc tcagctggga ggtgactctg
     7021 gcggctcaaa aagaaccgcc gacggcagcg aattcgagcc caagaagaag aggaaagtcg
     7081 tatacatgca tggcagcggc gccaccaact tcagcctgct gaagcaggcc ggcgacgtgg
     7141 aggagaaccc cggccccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     7201 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtct ggcgagggcg
     7261 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     7321 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
     7381 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     7441 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     7501 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     7561 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
     7621 ccgacaagca gaagaacggc atcaaggcga acttcaagat ccgccacaac atcgaggacg
     7681 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     7741 tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga
     7801 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     7861 acgagctgta caagtaagct agcctaccgg gtaggggagg cgcttttccc aaggcagtct
     7921 ggagcatgcg ctttagcagc cccgctgggc acttggcgct acacaagtgg cctctggcct
     7981 cgcacacatt ccacatccac cggtaggcgc caaccggctc cgttctttgg tggccccttc
     8041 gcgccacctt ctactcctcc cctagtcagg aagttccccc ccgccccgca gctcgcgtcg
     8101 tgcaggacgt gacaaatgga agtagcacgt ctcactagtc tcgtgcagat ggacagcacc
     8161 gctgagcaat ggaagcgggt aggcctttgg ggcagcggcc aatagcagct ttgctccttc
     8221 gctttctggg ctcagaggct gggaaggggt gggtccgggg gcgggctcag gggcgggctc
     8281 aggggcgggg cgggcgcccg aaggtcctcc ggaggcccgg cattctgcac gcttcaaaag
     8341 cgcacgtctg ccgcgctgtt ctcctcttcc tcatctccgg gcctttcgac ctgcagccca
     8401 agcttaccat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca
     8461 gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg
     8521 atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg
     8581 ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca
     8641 cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt
     8701 tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc
     8761 ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg
     8821 gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg
     8881 ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg
     8941 tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg
     9001 gtgcctgatt ctagaaagtc gacaatcaac ctctggatta caaaatttgt gaaagattga
     9061 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     9121 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     9181 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     9241 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     9301 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     9361 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
     9421 catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     9481 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     9541 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     9601 ccgcctcccc gcgtcgactt taagaccaat gacttacaag gcagctgtag atcttagcca
     9661 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatct
     9721 gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     9781 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     9841 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     9901 gtggaaaatc tctagcagta cgtatagtag ttcatgtcat cttattattc agtatttata
     9961 acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg
    10021 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
    10081 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg
    10141 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
    10201 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
    10261 cagaagtagt gaggaggctt ttttggaggc ctagggacgt acccaattcg ccctatagtg
    10321 agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
    10381 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
    10441 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg
    10501 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
    10561 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
    10621 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
    10681 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
    10741 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
    10801 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
    10861 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
    10921 cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg
    10981 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
    11041 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
    11101 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
    11161 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
    11221 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
    11281 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
    11341 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
    11401 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
    11461 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
    11521 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
    11581 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
    11641 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
    11701 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
    11761 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
    11821 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
    11881 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
    11941 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
    12001 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
    12061 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
    12121 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
    12181 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
    12241 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
    12301 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
    12361 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
    12421 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
    12481 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
    12541 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
    12601 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
    12661 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
    12721 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
    12781 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
    12841 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
    12901 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
    12961 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
    13021 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
    13081 atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta accctcacta
    13141 aagggaacaa aagctggagc tgcaagctta a
//
