LOCUS       sequence-408413-       17580 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral transfer plasmid containing CMV-driven expression 
            cassette encoding PE2 enzyme for prime editing..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 17580)
  AUTHORS   Bulcaen M, Kortleven P, Liu RB, Maule G, Dreano E, Kelly M, Ensinck 
            MM, Thierie S, Smits M, Ciciani M, Hatton A, Chevalier B, Ramalho 
            AS, Casadevall I Solvas X, Debyser Z, Vermeulen F, Gijsbers R, 
            Sermet-Gaudelus I, Cereseto A, Carlon MS
  TITLE     Prime editing functionally corrects cystic fibrosis-causing CFTR 
            mutations in human organoids and airway epithelial cells.
  JOURNAL   Cell Rep Med. 2024 May 21;5(5):101544. doi: 
            10.1016/j.xcrm.2024.101544. Epub 2024 May 1.
  PUBMED    38697102
REFERENCE   2  (bases 1 to 17580)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell Rep 
            Med."; date: "2024-05-21"; volume: "5(5):101544. doi"; pages: " 
            10.1016/j.xcrm.2024.101544. Epub 2024 May 1"
COMMENT     Sequence Label: Lenti-CMV-PE2-IRES-ZeoR
FEATURES             Location/Qualifiers
     source          1..17580
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1191..1210)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(1424..1443)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1543..1565
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1603..1621)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1689..1793
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1794..1862,1863..2654)
                     /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(2012..2031)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2825..3413
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3314..3333
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3567..3584
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3654..3674)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3659..3988
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3839..3974
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          5122..5187
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             5317..5337
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    5762..5896
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5799..5818)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5853..5872
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             6064..6697
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    6744..6869
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7362..7595
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7780..7824
                     /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    8117..8234
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        8293..8596
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8597..8800
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8751..8771
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8918..8937
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        8918..8936
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             8986..9006
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(9007..11520,11521..11523,11524..13107)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(H840A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the H840A mutation in the HNH catalytic domain"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             9007..13107
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             15280..15300
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    15443..15904
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(15499..15516)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     15726..15745
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             15912..16286
                     /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
                     EFALRDPAGNCVHFVADEQD"
     misc_feature    16305..16893
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(16358..16378)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(16776..16787)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             17063..17243
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
ORIGIN
        1 acacaagagg aaactgagag ggaaagatgt tgaagtaatt ttcccacaat tacagcatcc
       61 gttagttacg actctatgat cttctgacac aaattccatt tactcctcac cctatgactc
      121 agtcgaatat atcaaagtta tggacattat gctaagtaac aaattaccct tttatatagt
      181 aaatactgag tagattgaga gaagaaattg tttgcaaacc tgaatagctt caagaagaag
      241 agaagtgagg ataagaataa cagttgtcat ttaacaagtt ttaacaagta acttggttag
      301 aaagggattc aaatgcataa agcaagggat aaatttttct ggcaacaaga ctatacaata
      361 taaccttaaa tatgacttca aataattgtt ggaacttgat aaaactaatt aaatattatt
      421 gaagattatc aatattataa atgtaattta cttttaaaaa gggaacatag aaatgtgtat
      481 cattagagta gaaaacaatc cttattatca caatttgtca aaacaagttt gttattaaca
      541 caagtagaat actgcattca attaagttga ctgcagattt tgtgttttgt taaaattaga
      601 aagagataac aacaatttga attattgaaa gtaacatgta aatagttcta catacgttct
      661 tttgacatct tgttcaatca ttgatcgaag ttctttatct tggaagaatt tgttccaaag
      721 actctgaaat aaggaaaaca atctattata tagtctcaca cctttgtttt acttttagtg
      781 atttcaattt aataatgtaa atggttaaaa tttattcttc tctgagatca tttcacattg
      841 cagatagaaa acctgagact ggggtaattt ttattaaaat ctaatttaat ctcagaaaca
      901 catctttatt ctaacatcaa tttttccagt ttgatattat catataaagt cagccttcct
      961 catctgcagg ttccacaaca aaaatccaac caactgtgga tcaaaaatat tgggaaaaaa
     1021 ttaaaaatag caatacaaca ataaaaaaat acaaatcaga aaaacagcac agtataacaa
     1081 ctttatttag catttacaat ctattaggta ttataagtaa tctagccaga tcctctacgc
     1141 cggacgcatc gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc
     1201 cgacatcacc gatggggaag atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg
     1261 cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg ggcgccatct ccttgcatgc
     1321 accattcctt gcggcggcgg tgctcaacgg cctcaaccta ctactgggct gcttcctaat
     1381 gcaggagtcg cataagggag agcgtcgaat ggtgcactct cagtacaatc tgctctgatg
     1441 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     1501 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     1561 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     1621 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     1681 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     1741 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     1801 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     1861 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     1921 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     1981 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     2041 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     2101 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     2161 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     2221 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     2281 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     2341 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     2401 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     2461 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     2521 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     2581 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     2641 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     2701 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     2761 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     2821 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     2881 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     2941 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     3001 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     3061 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     3121 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     3181 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     3241 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     3301 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     3361 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     3421 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     3481 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     3541 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     3601 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tgtggaatgt
     3661 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3721 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     3781 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     3841 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     3901 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     3961 cttttttgga ggcctaggct tttgcaaaaa gcttggacac aagacaggct tgcgagatat
     4021 gtttgagaat accactttat cccgcgtcag ggagaggcag tgcgtaaaaa gacgcggact
     4081 catgtgaaat actggttttt agtgcgccag atctctataa tctcgcgcaa cctattttcc
     4141 cctcgaacac tttttaagcc gtagataaac aggctgggac acttcacatg agcgaaaaat
     4201 acatcgtcac ctgggacatg ttgcagatcc atgcacgtaa actcgcaagc cgactgatgc
     4261 cttctgaaca atggaaaggc attattgccg taagccgtgg cggtctgtac cgggtgcgtt
     4321 actggcgcgt gaactgggta ttcgtcatgt cgataccgtt tgtatttcca gctacgatca
     4381 cgacaaccag cgcgagctta aagtgctgaa acgcgcagaa ggcgatggcg aaggcttcat
     4441 cgttattgat gacctggtgg ataccggtgg tactgcggtt gcgattcgtg aaatgtatcc
     4501 aaaagcgcac tttgtcacca tcttcgcaaa accggctggt cgtccgctgg ttgatgacta
     4561 tgttgttgat atcccgcaag atacctggat tgaacagccg tgggatatgg gcgtcgtatt
     4621 cgtcccgcca atctccggtc gctaatcttt tcaacgcctg gcactgccgg gcgttgttct
     4681 ttttaacttc aggcgggtta caatagtttc cagtaagtat tctggaggct gcatccatga
     4741 cacaggcaaa cctgagcgaa accctgttca aaccccgctt taaacatcct gaaacctcga
     4801 cgctagtccg ccgctttaat cacggcgcac aaccgcctgt gcagtcggcc cttgatggta
     4861 aaaccatccc tcactggtat cgcatgatta accgtctgat gtggatctgg cgcggcattg
     4921 acccacgcga aatcctcgac gtccaggcac gtattgtgat gagcgatgcc gaacgtaccg
     4981 acgatgattt atacgatacg gtgattggct accgtggcgg caactggatt tatgagtggg
     5041 ccccggatct ttgtgaagga accttacttc tgtggtgtga cataattgga caaactacct
     5101 acagagattt aaagctctaa ggtaaatata aaatttttaa gtgtataatg tgttaaacta
     5161 ctgattctaa ttgtttgtgt attttagatt ccaacctatg gaactgatga atgggagcag
     5221 tggtggaatg cctttaatga ggaaaacctg ttttgctcag aagaaatgcc atctagtgat
     5281 gatgaggcta ctgctgactc tcaacattct actcctccaa aaaagaagag aaaggtagaa
     5341 gaccccaagg actttccttc agaattgcta agttttttga gtcatgctgt gtttagtaat
     5401 agaactcttg cttgctttgc tatttacacc acaaaggaaa aagctgcact gctatacaag
     5461 aaaattatgg aaaaatattc tgtaaccttt ataagtaggc ataacagtta taatcataac
     5521 atactgtttt ttcttactcc acacaggcat agagtgtctg ctattaataa ctatgctcaa
     5581 aaattgtgta cctttagctt tttaatttgt aaaggggtta ataaggaata tttgatgtat
     5641 agtgccttga ctagagatca taatcagcca taccacattt gtagaggttt tacttgcttt
     5701 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     5761 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     5821 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     5881 ttatcatgtc tggatcaact ggataactca agctaaccaa aatcatccca aacttcccac
     5941 cccataccct attaccactg ccaattacct gtggtttcat ttactctaaa cctgtgattc
     6001 ctctgaatta ttttcatttt aaagaaattg tatttgttaa atatgtacta caaacttagt
     6061 agttggaagg gctaattcac tcccaaagaa gacaagatat ccttgatctg tggatctacc
     6121 acacacaagg ctacttccct gattagcaga actacacacc agggccaggg gtcagatatc
     6181 cactgacctt tggatggtgc tacaagctag taccagttga gccagataag gtagaagagg
     6241 ccaataaagg agagaacacc agcttgttac accctgtgag cctgcatggg atggatgacc
     6301 cggagagaga agtgttagag tggaggtttg acagccgcct agcatttcat cacgtggccc
     6361 gagagctgca tccggagtac ttcaagaact gctgatatcg agcttgctac aagggacttt
     6421 ccgctgggga ctttccaggg aggcgtggcc tgggcgggac tggggagtgg cgagccctca
     6481 gatcctgcat ataagcagct gctttttgcc tgtactgggt ctctctggtt agaccagatc
     6541 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     6601 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     6661 ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga
     6721 aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca
     6781 cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct
     6841 agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat
     6901 gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat
     6961 gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag
     7021 gctgtagaca aatactggga cagctacaac catcccttca gacaggatca gaagaactta
     7081 gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag
     7141 acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac
     7201 agcaagcggc cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt
     7261 gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca
     7321 aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg
     7381 ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc
     7441 agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg
     7501 caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg
     7561 gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa
     7621 ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag
     7681 atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta
     7741 atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg
     7801 gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat
     7861 ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta
     7921 ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc
     7981 ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac
     8041 agagacagat ccattcgatt agtgaacgga tctcgacggt atcgccgaat tcacaaatgg
     8101 cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa
     8161 gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa
     8221 aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccactttgg ggcgataagc
     8281 ttgggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     8341 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     8401 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     8461 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     8521 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     8581 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     8641 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     8701 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     8761 cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga accgtcagat
     8821 cgcctggaga cgccatccac gctgttttga cctccataga agacaccgac tctagaggtt
     8881 tagtgaaccg tcagatccgc tagagatccg cggccgctaa tacgactcac tatagggaga
     8941 gccgccacca tgaaacggac agccgacgga agcgagttcg agtcaccaaa gaagaagcgg
     9001 aaagtcgaca agaagtacag catcggcctg gacatcggca ccaactctgt gggctgggcc
     9061 gtgatcaccg acgagtacaa ggtgcccagc aagaaattca aggtgctggg caacaccgac
     9121 cggcacagca tcaagaagaa cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc
     9181 gaggccaccc ggctgaagag aaccgccaga agaagataca ccagacggaa gaaccggatc
     9241 tgctatctgc aagagatctt cagcaacgag atggccaagg tggacgacag cttcttccac
     9301 agactggaag agtccttcct ggtggaagag gataagaagc acgagcggca ccccatcttc
     9361 ggcaacatcg tggacgaggt ggcctaccac gagaagtacc ccaccatcta ccacctgaga
     9421 aagaaactgg tggacagcac cgacaaggcc gacctgcggc tgatctatct ggccctggcc
     9481 cacatgatca agttccgggg ccacttcctg atcgagggcg acctgaaccc cgacaacagc
     9541 gacgtggaca agctgttcat ccagctggtg cagacctaca accagctgtt cgaggaaaac
     9601 cccatcaacg ccagcggcgt ggacgccaag gccatcctgt ctgccagact gagcaagagc
     9661 agacggctgg aaaatctgat cgcccagctg cccggcgaga agaagaatgg cctgttcgga
     9721 aacctgattg ccctgagcct gggcctgacc cccaacttca agagcaactt cgacctggcc
     9781 gaggatgcca aactgcagct gagcaaggac acctacgacg acgacctgga caacctgctg
     9841 gcccagatcg gcgaccagta cgccgacctg tttctggccg ccaagaacct gtccgacgcc
     9901 atcctgctga gcgacatcct gagagtgaac accgagatca ccaaggcccc cctgagcgcc
     9961 tctatgatca agagatacga cgagcaccac caggacctga ccctgctgaa agctctcgtg
    10021 cggcagcagc tgcctgagaa gtacaaagag attttcttcg accagagcaa gaacggctac
    10081 gccggctaca ttgacggcgg agccagccag gaagagttct acaagttcat caagcccatc
    10141 ctggaaaaga tggacggcac cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg
    10201 cggaagcagc ggaccttcga caacggcagc atcccccacc agatccacct gggagagctg
    10261 cacgccattc tgcggcggca ggaagatttt tacccattcc tgaaggacaa ccgggaaaag
    10321 atcgagaaga tcctgacctt ccgcatcccc tactacgtgg gccctctggc caggggaaac
    10381 agcagattcg cctggatgac cagaaagagc gaggaaacca tcaccccctg gaacttcgag
    10441 gaagtggtgg acaagggcgc ttccgcccag agcttcatcg agcggatgac caacttcgat
    10501 aagaacctgc ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga gtacttcacc
    10561 gtgtataacg agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc
    10621 ctgagcggcg agcagaaaaa ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg
    10681 accgtgaagc agctgaaaga ggactacttc aagaaaatcg agtgcttcga ctccgtggaa
    10741 atctccggcg tggaagatcg gttcaacgcc tccctgggca cataccacga tctgctgaaa
    10801 attatcaagg acaaggactt cctggacaat gaggaaaacg aggacattct ggaagatatc
    10861 gtgctgaccc tgacactgtt tgaggacaga gagatgatcg aggaacggct gaaaacctat
    10921 gcccacctgt tcgacgacaa agtgatgaag cagctgaagc ggcggagata caccggctgg
    10981 ggcaggctga gccggaagct gatcaacggc atccgggaca agcagtccgg caagacaatc
    11041 ctggatttcc tgaagtccga cggcttcgcc aacagaaact tcatgcagct gatccacgac
    11101 gacagcctga cctttaaaga ggacatccag aaagcccagg tgtccggcca gggcgatagc
    11161 ctgcacgagc acattgccaa tctggccggc agccccgcca ttaagaaggg catcctgcag
    11221 acagtgaagg tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc
    11281 gtgatcgaaa tggccagaga gaaccagacc acccagaagg gacagaagaa cagccgcgag
    11341 agaatgaagc ggatcgaaga gggcatcaaa gagctgggca gccagatcct gaaagaacac
    11401 cccgtggaaa acacccagct gcagaacgag aagctgtacc tgtactacct gcagaatggg
    11461 cgggatatgt acgtggacca ggaactggac atcaaccggc tgtccgacta cgatgtggac
    11521 gctatcgtgc ctcagagctt tctgaaggac gactccatcg acaacaaggt gctgaccaga
    11581 agcgacaaga accggggcaa gagcgacaac gtgccctccg aagaggtcgt gaagaagatg
    11641 aagaactact ggcggcagct gctgaacgcc aagctgatta cccagagaaa gttcgacaat
    11701 ctgaccaagg ccgagagagg cggcctgagc gaactggata aggccggctt catcaagaga
    11761 cagctggtgg aaacccggca gatcacaaag cacgtggcac agatcctgga ctcccggatg
    11821 aacactaagt acgacgagaa tgacaagctg atccgggaag tgaaagtgat caccctgaag
    11881 tccaagctgg tgtccgattt ccggaaggat ttccagtttt acaaagtgcg cgagatcaac
    11941 aactaccacc acgcccacga cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa
    12001 aagtacccta agctggaaag cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg
    12061 aagatgatcg ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta cttcttctac
    12121 agcaacatca tgaacttttt caagaccgag attaccctgg ccaacggcga gatccggaag
    12181 cggcctctga tcgagacaaa cggcgaaacc ggggagatcg tgtgggataa gggccgggat
    12241 tttgccaccg tgcggaaagt gctgagcatg ccccaagtga atatcgtgaa aaagaccgag
    12301 gtgcagacag gcggcttcag caaagagtct atcctgccca agaggaacag cgataagctg
    12361 atcgccagaa agaaggactg ggaccctaag aagtacggcg gcttcgacag ccccaccgtg
    12421 gcctattctg tgctggtggt ggccaaagtg gaaaagggca agtccaagaa actgaagagt
    12481 gtgaaagagc tgctggggat caccatcatg gaaagaagca gcttcgagaa gaatcccatc
    12541 gactttctgg aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat caagctgcct
    12601 aagtactccc tgttcgagct ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa
    12661 ctgcagaagg gaaacgaact ggccctgccc tccaaatatg tgaacttcct gtacctggcc
    12721 agccactatg agaagctgaa gggctccccc gaggataatg agcagaaaca gctgtttgtg
    12781 gaacagcaca agcactacct ggacgagatc atcgagcaga tcagcgagtt ctccaagaga
    12841 gtgatcctgg ccgacgctaa tctggacaaa gtgctgtccg cctacaacaa gcaccgggat
    12901 aagcccatca gagagcaggc cgagaatatc atccacctgt ttaccctgac caatctggga
    12961 gcccctgccg ccttcaagta ctttgacacc accatcgacc ggaagaggta caccagcacc
    13021 aaagaggtgc tggacgccac cctgatccac cagagcatca ccggcctgta cgagacacgg
    13081 atcgacctgt ctcagctggg aggtgactct ggaggatcta gcggaggatc ctctggcagc
    13141 gagacaccag gaacaagcga gtcagcaaca ccagagagca gtggcggcag cagcggcggc
    13201 agcagcaccc taaatataga agatgagtat cggctacatg agacctcaaa agagccagat
    13261 gtttctctag ggtccacatg gctgtctgat tttcctcagg cctgggcgga aaccgggggc
    13321 atgggactgg cagttcgcca agctcctctg atcatacctc tgaaagcaac ctctaccccc
    13381 gtgtccataa aacaataccc catgtcacaa gaagccagac tggggatcaa gccccacata
    13441 cagagactgt tggaccaggg aatactggta ccctgccagt ccccctggaa cacgcccctg
    13501 ctacccgtta agaaaccagg gactaatgat tataggcctg tccaggatct gagagaagtc
    13561 aacaagcggg tggaagacat ccaccccacc gtgcccaacc cttacaacct cttgagcggg
    13621 ctcccaccgt cccaccagtg gtacactgtg cttgatttaa aggatgcctt tttctgcctg
    13681 agactccacc ccaccagtca gcctctcttc gcctttgagt ggagagatcc agagatggga
    13741 atctcaggac aattgacctg gaccagactc ccacagggtt tcaaaaacag tcccaccctg
    13801 tttaatgagg cactgcacag agacctagca gacttccgga tccagcaccc agacttgatc
    13861 ctgctacagt acgtggatga cttactgctg gccgccactt ctgagctaga ctgccaacaa
    13921 ggtactcggg ccctgttaca aaccctaggg aacctcgggt atcgggcctc ggccaagaaa
    13981 gcccaaattt gccagaaaca ggtcaagtat ctggggtatc ttctaaaaga gggtcagaga
    14041 tggctgactg aggccagaaa agagactgtg atggggcagc ctactccgaa gacccctcga
    14101 caactaaggg agttcctagg gaaggcaggc ttctgtcgcc tcttcatccc tgggtttgca
    14161 gaaatggcag cccccctgta ccctctcacc aaaccgggga ctctgtttaa ttggggccca
    14221 gaccaacaaa aggcctatca agaaatcaag caagctcttc taactgcccc agccctgggg
    14281 ttgccagatt tgactaagcc ctttgaactc tttgtcgacg agaagcaggg ctacgccaaa
    14341 ggtgtcctaa cgcaaaaact gggaccttgg cgtcggccgg tggcctacct gtccaaaaag
    14401 ctagacccag tagcagctgg gtggccccct tgcctacgga tggtagcagc cattgccgta
    14461 ctgacaaagg atgcaggcaa gctaaccatg ggacagccac tagtcattct ggccccccat
    14521 gcagtagagg cactagtcaa acaacccccc gaccgctggc tttccaacgc ccggatgact
    14581 cactatcagg ccttgctttt ggacacggac cgggtccagt tcggaccggt ggtagccctg
    14641 aacccggcta cgctgctccc actgcctgag gaagggctgc aacacaactg ccttgatatc
    14701 ctggccgaag cccacggaac ccgacccgac ctaacggacc agccgctccc agacgccgac
    14761 cacacctggt acacggatgg aagcagtctc ttacaagagg gacagcgtaa ggcgggagct
    14821 gcggtgacca ccgagaccga ggtaatctgg gctaaagccc tgccagccgg gacatccgct
    14881 cagcgggctg aactgatagc actcacccag gccctaaaga tggcagaagg taagaagcta
    14941 aatgtttata ctgatagccg ttatgctttt gctactgccc atatccatgg agaaatatac
    15001 agaaggcgtg ggtggctcac atcagaaggc aaagagatca aaaataaaga cgagatcttg
    15061 gccctactaa aagccctctt tctgcccaaa agacttagca taatccattg tccaggacat
    15121 caaaagggac acagcgccga ggctagaggc aaccggatgg ctgaccaagc ggcccgaaag
    15181 gcagccatca cagagactcc agacacctct accctcctca tagaaaattc atcaccctct
    15241 ggcggctcaa aaagaaccgc cgacggcagc gaattcgagc ccaagaagaa gaggaaagtc
    15301 taatgtacaa gtaaactagt gctctagccc aattccgccc cctccccccc cctaacgtta
    15361 ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta ttttccacca
    15421 tattgccgtc ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca
    15481 ttcctagggg tctttcccct ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg
    15541 aagcagttcc tctggaagct tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc
    15601 agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt gtataagata
    15661 cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt gtggaaagag
    15721 tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag aaggtacccc
    15781 attgtatggg atctgatctg gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt
    15841 taaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg
    15901 ataatttcga aatggccaag ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg
    15961 ccggagcggt cgagttctgg accgaccggc tcgggttctc ccgggacttc gtggaggacg
    16021 acttcgccgg tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc caggaccagg
    16081 tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg
    16141 agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc atgaccgaga
    16201 tcggcgagca gccgtggggg cgggagttcg ccctgcgcga cccggccggc aactgcgtgc
    16261 acttcgtggc cgacgagcag gactgaacgc gttgaattcc cgataatcaa cctctggatt
    16321 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
    16381 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
    16441 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
    16501 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
    16561 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
    16621 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
    16681 ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct
    16741 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
    16801 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
    16861 cgagtcggat ctccctttgg gccgcctccc cgcatcggga attctgtcga ctggtacctt
    16921 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg
    16981 gactggaagg gctaattcac tcccaacgaa gacaaaatcg tcgagagatg ctgcatataa
    17041 gcagctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct
    17101 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
    17161 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
    17221 gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata
    17281 acttgcaaag aaatgaatat cagagagtga gaggccttga cattataata gatttagcag
    17341 gaattgaact aggagtggag cacacaggca aagctgcaga agtacttgga agaagccacc
    17401 agagatactc acgattctgc acatacctgg ctaatcccag atcctaagga ttacattaag
    17461 tttactaaca tttatataat gatttatagt ttaaagtata aacttatcta atttactatt
    17521 ctgacagata ttaattaatc ctcaaatatc ataagagatg attactatta tccccattta
//
