LOCUS       sequence-361796-       16482 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Single EF1a-driven PE2-P2A-hMLH1dn (Addgene #174828) and PaqCI 
            restriction sites for pegRNA cloning. Lentiviral backbone and AmpR..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16482)
  AUTHORS   Dirkx N, Weuring WJ, De Vriendt E, Smal N, van de Vondervoort J, van
            't Slot R, Koetsier M, Zonnekein N, De Pooter T, Weckhuysen S, 
            Koeleman BPC
  TITLE     Increased prime edit rates in KCNQ2 and SCN1A via single nicking 
            all-in-one plasmids.
  JOURNAL   BMC Biol. 2023 Jul 13;21(1):156. doi: 10.1186/s12915-023-01646-7.
  PUBMED    37443005
REFERENCE   2  (bases 1 to 16482)
  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: "BMC Biol.";
            date: "2023-07-13"; volume: "21(1):156. doi"; pages: " 
            10.1186/s12915-023-01646-7"
COMMENT     Sequence Label: pLenti_Ef1a:PE2-P2A-hMLH1dn
FEATURES             Location/Qualifiers
     source          1..16482
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(252..268)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(252..268)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(265..287)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    276..292
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(300..306,307..324,325..330))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    345..366
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(483..500)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(654..1242)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(734..753)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1413..2204,2205..2273))
                     /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     2036..2055
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2274..2378)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    2426..2560
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2463..2482)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2517..2536
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             2728..3361
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    3408..3533
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4030..4263
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4448..4492
                     /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    4755..4870
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     promoter        5160..6342
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          5391..6333
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     6290..6310
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     6406..6425
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6406..6424
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6474..6494
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6495..10595
                     /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
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRKLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLKREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             10653..10673
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             12774..12794
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             12804..12830
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             12840..12896
                     /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"
     misc_feature    15178..15766
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(15231..15251)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(15649..15660)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             join(15973..16482,1..124)
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
ORIGIN
        1 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
       61 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
      121 agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata
      181 tcagagagtg agaggccttg acattgctag cgtttaccgt cgacctctag ctagagcttg
      241 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
      301 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
      361 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
      421 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
      481 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
      541 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
      601 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
      661 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
      721 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
      781 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
      841 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
      901 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
      961 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     1021 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     1081 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     1141 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     1201 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     1261 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     1321 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     1381 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     1441 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     1501 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     1561 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     1621 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     1681 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     1741 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     1801 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     1861 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     1921 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     1981 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     2041 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     2101 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     2161 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     2221 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     2281 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     2341 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     2401 cctgacgtcg acggatcggg agatcaactt gtttattgca gcttataatg gttacaaata
     2461 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     2521 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc aactggataa ctcaagctaa
     2581 ccaaaatcat cccaaacttc ccaccccata ccctattacc actgccaatt acctgtggtt
     2641 tcatttactc taaacctgtg attcctctga attattttca ttttaaagaa attgtatttg
     2701 ttaaatatgt actacaaact tagtagttgg aagggctaat tcactcccaa agaagacaag
     2761 atatccttga tctgtggatc taccacacac aaggctactt ccctgattag cagaactaca
     2821 caccagggcc aggggtcaga tatccactga cctttggatg gtgctacaag ctagtaccag
     2881 ttgagccaga taaggtagaa gaggccaata aaggagagaa caccagcttg ttacaccctg
     2941 tgagcctgca tgggatggat gacccggaga gagaagtgtt agagtggagg tttgacagcc
     3001 gcctagcatt tcatcacgtg gcccgagagc tgcatccgga gtacttcaag aactgctgat
     3061 atcgagcttg ctacaaggga ctttccgctg gggactttcc agggaggcgt ggcctgggcg
     3121 ggactgggga gtggcgagcc ctcagatcct gcatataagc agctgctttt tgcctgtact
     3181 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     3241 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     3301 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     3361 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca
     3421 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc
     3481 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta
     3541 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa
     3601 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc
     3661 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc
     3721 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg
     3781 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc
     3841 aaaacaaaag taagaccacc gcacagcaag cggccggccg ctgatcttca gacctggagg
     3901 aggagatatg agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga
     3961 accattagga gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc
     4021 agtgggaata ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc
     4081 agcgtcaatg acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca
     4141 gaacaatttg ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg
     4201 catcaagcag ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct
     4261 cctggggatt tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc
     4321 tagttggagt aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga
     4381 cagagaaatt aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca
     4441 gcaagaaaag aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg
     4501 gtttaacata acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt
     4561 ggtaggttta agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata
     4621 ttcaccatta tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg
     4681 aatagaagaa gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc
     4741 tcgacggtat cgcctttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa
     4801 tagtagacat aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa
     4861 ttcaaaattt tcgggtttat tacagggaca gcagagatcc agtttatcga tggtccgatt
     4921 tcagcaggtg atgaggttag cagtctcacc tgcgggcaga tcgaacctgc aggttccact
     4981 gggacgatta catgcatctg gatcgatgag taattcatac aaaaggactc gcccctgcct
     5041 tggggaatcc cagggaccgt cgttaaactc ccactaacgt agaacccaga gatcgctgcg
     5101 ttcccgcccc ctcacccgcc cgctctcgtc atcactgagg tggagaagag catgcgtgag
     5161 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg
     5221 gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg
     5281 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag
     5341 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg
     5401 tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta
     5461 cttccacgcc cctggctgca gtacgtgatt cttgatcccg agcttcgggt tggaagtggg
     5521 tgggagagtt cgaggccttg cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc
     5581 tggcttgggc gctggggccg ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct
     5641 gctttcgata agtctctagc catttaaaat ttttgatgac ctgctgcgac gctttttttc
     5701 tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca ctggtatttc ggtttttggg
     5761 gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg
     5821 cgagcgcggc caccgagaat cggacggggg tagtctcaag ctggccggcc tgctctggtg
     5881 cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg caaggctggc ccggtcggca
     5941 ccagttgcgt gagcggaaag atggccgctt cccggccctg ctgcagggag ctcaaaatgg
     6001 aggacgcggc gctcgggaga gcgggcgggt gagtcaccca cacaaaggaa aagggccttt
     6061 ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc gggcgccgtc caggcacctc
     6121 gattagttct cgagcttttg gagtacgtcg tctttaggtt ggggggaggg gttttatgcg
     6181 atggagtttc cccacactga gtgggtggag actgaagtta ggccagcttg gcacttgatg
     6241 taattctcct tggaatttgc cctttttgag tttggatctt ggttcattct caagcctcag
     6301 acagtggttc aaagtttttt tcttccattt caggtgtcgt gaggatctat ttccggtgaa
     6361 ttggtttaaa cgcgatcgcc tcgagggatc ctctagagcg gccgctaata cgactcacta
     6421 tagggagagc cgccaccatg aaacggacag ccgacggaag cgagttcgag tcaccaaaga
     6481 agaagcggaa agtcgacaag aagtacagca tcggcctgga catcggcacc aactctgtgg
     6541 gctgggccgt gatcaccgac gagtacaagg tgcccagcaa gaaattcaag gtgctgggca
     6601 acaccgaccg gcacagcatc aagaagaacc tgatcggagc cctgctgttc gacagcggcg
     6661 aaacagccga ggccacccgg ctgaagagaa ccgccagaag aagatacacc agacggaaga
     6721 accggatctg ctatctgcaa gagatcttca gcaacgagat ggccaaggtg gacgacagct
     6781 tcttccacag actggaagag tccttcctgg tggaagagga taagaagcac gagcggcacc
     6841 ccatcttcgg caacatcgtg gacgaggtgg cctaccacga gaagtacccc accatctacc
     6901 acctgagaaa gaaactggtg gacagcaccg acaaggccga cctgcggctg atctatctgg
     6961 ccctggccca catgatcaag ttccggggcc acttcctgat cgagggcgac ctgaaccccg
     7021 acaacagcga cgtggacaag ctgttcatcc agctggtgca gacctacaac cagctgttcg
     7081 aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc catcctgtct gccagactga
     7141 gcaagagcag aaagctggaa aatctgatcg cccagctgcc cggcgagaag aagaatggcc
     7201 tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc caacttcaag agcaacttcg
     7261 acctggccga ggatgccaaa ctgcagctga gcaaggacac ctacgacgac gacctggaca
     7321 acctgctggc ccagatcggc gaccagtacg ccgacctgtt tctggccgcc aagaacctgt
     7381 ccgacgccat cctgctgagc gacatcctga gagtgaacac cgagatcacc aaggcccccc
     7441 tgagcgcctc tatgatcaag agatacgacg agcaccacca ggacctgacc ctgctgaaag
     7501 ctctcgtgcg gcagcagctg cctgagaagt acaaagagat tttcttcgac cagagcaaga
     7561 acggctacgc cggctacatt gacggcggag ccagccagga agagttctac aagttcatca
     7621 agcccatcct ggaaaagatg gacggcaccg aggaactgct cgtgaagctg aagagagagg
     7681 acctgctgcg gaagcagcgg accttcgaca acggcagcat cccccaccag atccacctgg
     7741 gagagctgca cgccattctg cggcggcagg aagattttta cccattcctg aaggacaacc
     7801 gggaaaagat cgagaagatc ctgaccttcc gcatccccta ctacgtgggc cctctggcca
     7861 ggggaaacag cagattcgcc tggatgacca gaaagagcga ggaaaccatc accccctgga
     7921 acttcgagga agtggtggac aagggcgctt ccgcccagag cttcatcgag cggatgacca
     7981 acttcgataa gaacctgccc aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt
     8041 acttcaccgt gtataacgag ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc
     8101 ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc
     8161 ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag tgcttcgact
     8221 ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca taccacgatc
     8281 tgctgaaaat tatcaaggac aaggacttcc tggacaatga ggaaaacgag gacattctgg
     8341 aagatatcgt gctgaccctg acactgtttg aggacagaga gatgatcgag gaacggctga
     8401 aaacctatgc ccacctgttc gacgacaaag tgatgaagca gctgaagcgg cggagataca
     8461 ccggctgggg caggctgagc cggaagctga tcaacggcat ccgggacaag cagtccggca
     8521 agacaatcct ggatttcctg aagtccgacg gcttcgccaa cagaaacttc atgcagctga
     8581 tccacgacga cagcctgacc tttaaagagg acatccagaa agcccaggtg tccggccagg
     8641 gcgatagcct gcacgagcac attgccaatc tggccggcag ccccgccatt aagaagggca
     8701 tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg
     8761 agaacatcgt gatcgaaatg gccagagaga accagaccac ccagaaggga cagaagaaca
     8821 gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc cagatcctga
     8881 aagaacaccc cgtggaaaac acccagctgc agaacgagaa gctgtacctg tactacctgc
     8941 agaatgggcg ggatatgtac gtggaccagg aactggacat caaccggctg tccgactacg
     9001 atgtggacgc tatcgtgcct cagagctttc tgaaggacga ctccatcgac aacaaggtgc
     9061 tgaccagaag cgacaagaac cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga
     9121 agaagatgaa gaactactgg cggcagctgc tgaacgccaa gctgattacc cagagaaagt
     9181 tcgacaatct gaccaaggcc gagagaggcg gcctgagcga actggataag gccggcttca
     9241 tcaagagaca gctggtggaa acccggcaga tcacaaagca cgtggcacag atcctggact
     9301 cccggatgaa cactaagtac gacgagaatg acaagctgat ccgggaagtg aaagtgatca
     9361 ccctgaagtc caagctggtg tccgatttcc ggaaggattt ccagttttac aaagtgcgcg
     9421 agatcaacaa ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc
     9481 tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta cggcgactac aaggtgtacg
     9541 acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg caaggctacc gccaagtact
     9601 tcttctacag caacatcatg aactttttca agaccgagat taccctggcc aacggcgaga
     9661 tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg
     9721 gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa
     9781 agaccgaggt gcagacaggc ggcttcagca aagagtctat cctgcccaag aggaacagcg
     9841 ataagctgat cgccagaaag aaggactggg accctaagaa gtacggcggc ttcgacagcc
     9901 ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac
     9961 tgaagagtgt gaaagagctg ctggggatca ccatcatgga aagaagcagc ttcgagaaga
    10021 atcccatcga ctttctggaa gccaagggct acaaagaagt gaaaaaggac ctgatcatca
    10081 agctgcctaa gtactccctg ttcgagctgg aaaacggccg gaagagaatg ctggcctctg
    10141 ccggcgaact gcagaaggga aacgaactgg ccctgccctc caaatatgtg aacttcctgt
    10201 acctggccag ccactatgag aagctgaagg gctcccccga ggataatgag cagaaacagc
    10261 tgtttgtgga acagcacaag cactacctgg acgagatcat cgagcagatc agcgagttct
    10321 ccaagagagt gatcctggcc gacgctaatc tggacaaagt gctgtccgcc tacaacaagc
    10381 accgggataa gcccatcaga gagcaggccg agaatatcat ccacctgttt accctgacca
    10441 atctgggagc ccctgccgcc ttcaagtact ttgacaccac catcgaccgg aagaggtaca
    10501 ccagcaccaa agaggtgctg gacgccaccc tgatccacca gagcatcacc ggcctgtacg
    10561 agacacggat cgacctgtct cagctgggag gtgactccgg cggaagctct ggtggcagca
    10621 agcggaccgc cgacggctct gaattcgaga gccctaagaa gaaaagaaag gtgagcggag
    10681 gctctagcgg cggaagcacc ctgaacattg aagacgagta tagactgcat gaaacaagca
    10741 aggaacccga cgtgtccctg ggctccacct ggctgtccga ctttccccag gcctgggccg
    10801 agacaggagg aatgggcctg gccgtgcggc aggcacccct gatcatccct ctgaaggcca
    10861 cctctacacc cgtgagcatc aagcagtacc ctatgtctca ggaggccaga ctgggcatca
    10921 agcctcacat ccagaggctg ctggaccagg gcatcctggt gccatgccag agcccctgga
    10981 acacaccact gctgcccgtg aagaagccag gcaccaatga ctatagaccc gtgcaggatc
    11041 tgagagaggt gaacaagagg gtggaggata tccaccccac cgtgcccaac ccttacaatc
    11101 tgctgtccgg cctgccccct tctcaccagt ggtatacagt gctggacctg aaggatgcct
    11161 tcttttgtct gagactgcac cctaccagcc agccactgtt cgcctttgag tggagggacc
    11221 ctgagatggg catctctggc cagctgacct ggacacgcct gcctcagggc ttcaagaata
    11281 gcccaacact gtttaacgag gccctgcacc gcgacctggc agatttccgg atccagcacc
    11341 cagatctgat cctgctgcag tacgtggacg atctgctgct ggccgccacc agcgagctgg
    11401 attgccagca gggaacacgc gccctgctgc agaccctggg aaacctggga tatagggcat
    11461 ccgccaagaa ggcccagatc tgtcagaagc aggtgaagta cctgggctat ctgctgaagg
    11521 agggccagag atggctgaca gaggccagga aggagacagt gatgggccag ccaacaccca
    11581 agaccccaag acagctgagg gagttcctgg gcaaagcagg attttgcagg ctgttcatcc
    11641 caggattcgc agagatggca gcacctctgt acccactgac caagccgggc accctgttta
    11701 attggggccc tgaccagcag aaggcctatc aggagatcaa gcaggccctg ctgacagcac
    11761 cagccctggg cctgccagac ctgaccaagc ctttcgagct gtttgtggat gagaagcagg
    11821 gctacgccaa gggcgtgctg acccagaagc tgggaccatg gagacggccc gtggcctatc
    11881 tgtccaagaa gctggaccca gtggcagcag gatggccacc atgcctgagg atggtggcag
    11941 caatcgccgt gctgacaaag gatgccggca agctgaccat gggacagcca ctggtcatcc
    12001 tggcaccaca cgcagtggag gccctggtga agcagcctcc agatcgctgg ctgtctaacg
    12061 cccggatgac acactaccag gccctgctgc tggacaccga tcgcgtgcag tttggccctg
    12121 tggtggccct gaatccagcc accctgctgc ctctgccaga ggagggcctg cagcacaact
    12181 gtctggacat cctggcagag gcacacggaa caaggccaga cctgaccgat cagcccctgc
    12241 ctgacgccga tcacacatgg tataccgatg gaagctccct gctgcaggag ggccagagga
    12301 aggcaggagc agcagtgacc acagagacag aagtgatctg ggccaaggcc ctgccagcag
    12361 gcacatccgc ccagcgggcc gagctgatcg ccctgaccca ggccctgaag atggccgagg
    12421 gcaagaagct gaacgtgtac acagactcca gatatgcctt cgccaccgca cacatccacg
    12481 gagagatcta caggcgccgg ggctggctga cctctgaggg caaggagatc aagaacaagg
    12541 atgagatcct ggccctgctg aaggccctgt ttctgcccaa gcggctgagc atcatccact
    12601 gtcctggaca ccagaaggga cactccgccg aggcaagggg caatcggatg gccgaccagg
    12661 ccgccagaaa ggctgctatt actgaaactc ccgacacttc cactctgctg attgaaaact
    12721 cctccccttc tggcggctca aaaagaaccg ccgacggcag cgaattcgag tctcccaaga
    12781 agaagaggaa agtcggctct ggccctgccg ctaagagagt gaagctggac ggaagcggag
    12841 ctactaactt cagcctgctg aagcaggctg gagacgtgga ggagaaccct ggacctagct
    12901 tcgttgctgg agtcatccgg agactggacg agacagtggt gaacagaatt gccgccggcg
    12961 aggtgatcca gagacctgcc aatgcaataa aggagatgat cgagaactgt ctggacgcca
    13021 agtccacaag cattcaggtg atcgtgaagg agggcggact gaagctgatc cagatccaag
    13081 acaacggcac aggcatcaga aaggaagatc tggacatcgt gtgtgaacgg ttcaccacat
    13141 ctaagctgca gtcttttgag gatctggcct ctatcagtac ctacggcttc agaggcgagg
    13201 ccctggccag catcagccac gtggcccatg tgaccatcac caccaaaacc gccgacggca
    13261 aatgcgctta tcgcgctagc tacagcgacg gcaagctgaa agccccgcca aagccttgcg
    13321 ccggcaacca gggtacacag ataacagtgg aggatctgtt ctacaacatc gccacccgga
    13381 gaaaggccct gaaaaatccc agcgaggagt acggcaagat cctggaagtc gtcggcagat
    13441 actccgtgca caacgccgga atcagcttta gcgtaaagaa gcagggagaa accgtggccg
    13501 atgtgcgcac cctgccaaat gccagcaccg tggataacat cagaagcatt ttcggaaatg
    13561 ccgtgtccag agaactgatc gagatcggct gcgaagataa gaccctggct tttaagatga
    13621 acggctacat cagcaacgcc aattactctg tgaagaagtg catctttctt ctgttcatca
    13681 accacagact ggtggaaagc accagcctgc ggaaagccat cgagacagtg tacgccgcct
    13741 acctgcctaa gaacacccac cccttcctgt acctgagcct cgagatcagc cctcagaacg
    13801 tggacgtcaa tgtgcatcct acaaagcacg aggtgcactt cctgcacgag gaaagcatcc
    13861 tggaaagagt gcagcagcac attgagagca agctgctggg ctctaacagc agcagaatgt
    13921 acttcacaca gaccctgctg cctggcctgg ccggcccctc aggcgaaatg gttaagtcca
    13981 caacctctct gacctcatcc agcaccagcg gttcttccga taaggtgtac gcccaccaga
    14041 tggtgcggac cgactctcgg gagcagaagc tggacgcctt tctgcaacct ctgagcaaac
    14101 ctctgagctc tcagcctcag gccatcgtga ccgaggacaa gacagatatc tcctccggcc
    14161 gtgccagaca gcaggacgaa gaaatgctcg agctgccagc tcctgccgag gtggccgcca
    14221 agaaccagag cctggaggga gataccacaa agggcaccag cgaaatgagc gagaagcggg
    14281 gccctacctc cagcaacccc agaaaacggc accgggagga cagcgacgtg gaaatggtgg
    14341 aggacgacag cagaaaggaa atgacagccg cttgtacccc tagaagaaga atcatcaacc
    14401 tgacctccgt gctgagcctg caggaggaga tcaacgagca gggccacgag gtgctgagag
    14461 agatgctgca caatcacagc ttcgtgggct gcgtgaaccc tcaatgggcc ctggctcagc
    14521 atcaaacaaa gctgtacctg ctgaacacca ccaagctgag cgaagagctg ttctaccaga
    14581 tcctcatcta cgacttcgcc aacttcggcg tgctacgcct gagcgagccc gcccctctgt
    14641 ttgacctggc catgctggct ctggatagcc cagaaagcgg ctggacagaa gaggacggac
    14701 ctaaagaggg gctggctgaa tacatcgtgg agttcctgaa gaaaaaggcc gagatgctgg
    14761 ccgactactt ttctctggaa atcgacgagg aaggcaacct gatcggcctg cctctgctga
    14821 tcgataacta cgtgcctccc ctggaaggcc tgcccatctt catcctgaga ctggctacag
    14881 aggtgaactg ggacgaggaa aaggaatgct tcgagtctct gagcaaggag tgcgccatgt
    14941 tctatagcat cagaaaacag tacatctctg aagagagcac tctgtctggc cagcagagtg
    15001 aagtgcccgg aagcatcccc aacagctgga agtggaccgt ggaacacatc gtgtacaagg
    15061 ccctgcggag ccacattctc cctcctaagc acttcaccga ggacggcaac atcctgcagc
    15121 tggccaacct gcccgacctt tataaggttt tctaaccggt tcagacgcgt ctggaacaat
    15181 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
    15241 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
    15301 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
    15361 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
    15421 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
    15481 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
    15541 ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc
    15601 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
    15661 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
    15721 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg aattaattct
    15781 gcagtcgaga cctagaaaaa catggagcaa tcacaagtag caatacagca gctaccaatg
    15841 ctgattgtgc ctggctagaa gcacaagagg aggaggaggt gggttttcca gtcacacctc
    15901 aggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag
    15961 aaaagagggg actggaaggg ctaattcact cccaacgaag acaagatatc cttgatctgt
    16021 ggatctacca cacacaaggc tacttccctg attagcagaa ctacacacca gggccagggg
    16081 tcagatatcc actgaccttt ggatggtgct acaagctagt accagttgag ccagataagg
    16141 tagaagaggc caataaagga gagaacacca gcttgttaca ccctgtgagc ctgcatggga
    16201 tggatgaccc ggagagagaa gtgttagagt ggaggtttga cagccgccta gcatttcatc
    16261 acgtggcccg agagctgcat ccggagtact tcaagaactg ctgatatcga gcttgctaca
    16321 agggactttc cgctggggac tttccaggga ggcgtggcct gggcgggact ggggagtggc
    16381 gagccctcag atcctgcata taagcagctg ctttttgcct gtactgggtc tctctggtta
    16441 gaccagatct gagcctggga gctctctggc taactaggga ac
//
