LOCUS       sequence-366206-        9540 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tier-1 vector encoding PhCMV-driven dCas9-3xNLS-VP64 expression 
            (PhCMV-dCas9-3xNLS-VP64-pA)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9540)
  AUTHORS   Haellman V, Strittmatter T, Bertschi A, Stucheli P, Fussenegger M
  TITLE     A versatile plasmid architecture for mammalian synthetic biology 
            (VAMSyB).
  JOURNAL   Metab Eng. 2021 Jul;66:41-50. doi: 10.1016/j.ymben.2021.04.003. Epub
            2021 Apr 18.
  PUBMED    33857582
REFERENCE   2  (bases 1 to 9540)
  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: "Metab 
            Eng."; date: "2021-07"; volume: "66:41-50. doi"; pages: " 
            10.1016/j.ymben.2021.04.003. Epub 2021 Apr 18"
COMMENT     Sequence Label: pVH333-1-Tier1-PhCMV-dCas9-3xNLS-VP64
FEATURES             Location/Qualifiers
     source          1..9540
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     66..86
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     160..179
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        160..178
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(219..242,243..245,246..2732,2733..2735,2736..4319)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4338..4358
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4362..4382
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4386..4406
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4413..4562
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     primer_bind     complement(4623..4640)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4629..4853
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4899..5327
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4986..5005)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5196..5217
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5336..5356)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5341..5670
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5521..5656
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5583..5602
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5737..6531
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5791..5810)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6401..6420
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6705..6826
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6742..6761)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6796..6815
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6875..6891)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6875..6891)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6888..6910)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6899..6915
                     /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(6923..6929,6930..6947,6948..6953))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6968..6989
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7106..7123)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7277..7865)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7357..7376)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8036..8827,8828..8896))
                     /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     8659..8678
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8897..9001)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        9072..9451
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(9452..9540,1..115)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
       61 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctctg
      121 gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc actataggga
      181 gacccaagct gttcgaagcg gaattcacca tgactagtga caagaagtat tctatcggac
      241 tggccatcgg gactaatagc gtcgggtggg ccgtgatcac tgacgagtac aaggtgccct
      301 ctaagaagtt caaggtgctc gggaacaccg accggcattc catcaagaaa aatctgatcg
      361 gagctctcct ctttgattca ggggagaccg ctgaagcaac ccgcctcaag cggactgcta
      421 gacggcggta caccaggagg aagaaccgga tttgttacct tcaagagata ttctccaacg
      481 aaatggcaaa ggtcgacgac agcttcttcc ataggctgga agaatcattc ctcgtggaag
      541 aggataagaa gcatgaacgg catcccatct tcggtaatat cgtcgacgag gtggcctatc
      601 acgagaaata cccaaccatc taccatcttc gcaaaaagct ggtggactca accgacaagg
      661 cagacctccg gcttatctac ctggccctgg cccacatgat caagttcaga ggccacttcc
      721 tgatcgaggg cgacctcaat cctgacaata gcgatgtgga taaactgttc atccagctgg
      781 tgcagactta caaccagctc tttgaagaga accccatcaa tgcaagcgga gtcgatgcca
      841 aggccattct gtcagcccgg ctgtcaaaga gccgcagact tgagaatctt atcgctcagc
      901 tgccgggtga aaagaaaaat ggactgttcg ggaacctgat tgctctttca cttgggctga
      961 ctcccaattt caagtctaat ttcgacctgg cagaggatgc caagctgcaa ctgtccaagg
     1021 acacctatga tgacgatctc gacaacctcc tggcccagat cggtgaccaa tacgccgacc
     1081 ttttccttgc tgctaagaat ctttctgacg ccatcctgct gtctgacatt ctccgcgtga
     1141 acactgaaat caccaaggcc cctctttcag cttcaatgat taagcggtat gatgagcacc
     1201 accaggacct gaccctgctt aaggcactcg tccggcagca gcttccggag aagtacaagg
     1261 aaatcttctt tgaccagtca aagaatggat acgccggcta catcgacgga ggtgcctccc
     1321 aagaggaatt ttataagttt atcaaaccta tccttgagaa gatggacggc accgaagagc
     1381 tcctcgtgaa actgaatcgg gaggatctgc tgcggaagca gcgcactttc gacaatggga
     1441 gcattcccca ccagatccat cttggggagc ttcacgccat ccttcggcgc caagaggact
     1501 tctacccctt tcttaaggac aacagggaga agattgagaa aattctcact ttccgcatcc
     1561 cctactacgt gggacccctc gccagaggaa atagccggtt tgcttggatg accagaaagt
     1621 cagaagaaac tatcactccc tggaacttcg aagaggtggt ggacaaggga gccagcgctc
     1681 agtcattcat cgaacggatg actaacttcg ataagaacct ccccaatgag aaggtcctgc
     1741 cgaaacattc cctgctctac gagtacttta ccgtgtacaa cgagctgacc aaggtgaaat
     1801 atgtcaccga agggatgagg aagcccgcat tcctgtcagg cgaacaaaag aaggcaattg
     1861 tggaccttct gttcaagacc aatagaaagg tgaccgtgaa gcagctgaag gaggactatt
     1921 tcaagaaaat tgaatgcttc gactctgtgg agattagcgg ggtcgaagat cggttcaacg
     1981 caagcctggg tacctaccat gatctgctta agatcatcaa ggacaaggat tttctggaca
     2041 atgaggagaa cgaggacatc cttgaggaca ttgtcctgac tctcactctg ttcgaggacc
     2101 gggaaatgat cgaggagagg cttaagacct acgcccatct gttcgacgat aaagtgatga
     2161 agcaacttaa acggagaaga tataccggat ggggacgcct tagccgcaaa ctcatcaacg
     2221 gaatccggga caaacagagc ggaaagacca ttcttgattt ccttaagagc gacggattcg
     2281 ctaatcgcaa cttcatgcaa cttatccatg atgattccct gacctttaag gaggacatcc
     2341 agaaggccca agtgtctgga caaggtgact cactgcacga gcatatcgca aatctggctg
     2401 gttcacccgc tattaagaag ggtattctcc agaccgtgaa agtcgtggac gagctggtca
     2461 aggtgatggg tcgccataaa ccagagaaca ttgtcatcga gatggccagg gaaaaccaga
     2521 ctacccagaa gggacagaag aacagcaggg agcggatgaa aagaattgag gaagggatta
     2581 aggagctcgg gtcacagatc cttaaagagc acccggtgga aaacacccag cttcagaatg
     2641 agaagctcta tctgtactac cttcaaaatg gacgcgatat gtatgtggac caagagcttg
     2701 atatcaacag gctctcagac tacgacgtgg acgccatcgt ccctcagagc ttcctcaaag
     2761 acgactcaat tgacaataag gtgctgactc gctcagacaa gaaccgggga aagtcagata
     2821 acgtgccctc agaggaagtc gtgaaaaaga tgaagaacta ttggcgccag cttctgaacg
     2881 caaagctgat cactcagcgg aagttcgaca atctcactaa ggctgagagg ggcggactga
     2941 gcgaactgga caaagcagga ttcattaaac ggcaacttgt ggagactcgg cagattacta
     3001 aacatgtcgc ccaaatcctt gactcacgca tgaataccaa gtacgacgaa aacgacaaac
     3061 ttatccgcga ggtgaaggtg attaccctga agtccaagct ggtcagcgat ttcagaaagg
     3121 actttcaatt ctacaaagtg cgggagatca ataactatca tcatgctcat gacgcatatc
     3181 tgaatgccgt ggtgggaacc gccctgatca agaagtaccc aaagctggaa agcgagttcg
     3241 tgtacggaga ctacaaggtc tacgacgtgc gcaagatgat tgccaaatct gagcaggaga
     3301 tcggaaaggc caccgcaaag tacttcttct acagcaacat catgaatttc ttcaagaccg
     3361 aaatcaccct tgcaaacggt gagatccgga agaggccgct catcgagact aatggggaga
     3421 ctggcgaaat cgtgtgggac aagggcagag atttcgctac cgtgcgcaaa gtgctttcta
     3481 tgcctcaagt gaacatcgtg aagaaaaccg aggtgcaaac cggaggcttt tctaaggaat
     3541 caatcctccc caagcgcaac tccgacaagc tcattgcaag gaagaaggat tgggacccta
     3601 agaagtacgg cggattcgat tcaccaactg tggcttattc tgtcctggtc gtggctaagg
     3661 tggaaaaagg aaagtctaag aagctcaaga gcgtgaagga actgctgggt atcaccatta
     3721 tggagcgcag ctccttcgag aagaacccaa ttgactttct cgaagccaaa ggttacaagg
     3781 aagtcaagaa ggaccttatc atcaagctcc caaagtatag cctgttcgaa ctggagaatg
     3841 ggcggaagcg gatgctcgcc tccgctggcg aacttcagaa gggtaatgag ctggctctcc
     3901 cctccaagta cgtgaatttc ctctaccttg caagccatta cgagaagctg aaggggagcc
     3961 ccgaggacaa cgagcaaaag caactgtttg tggagcagca taagcattat ctggacgaga
     4021 tcattgagca gatttccgag ttttctaaac gcgtcattct cgctgatgcc aacctcgata
     4081 aagtccttag cgcatacaat aagcacagag acaaaccaat tcgggagcag gctgagaata
     4141 tcatccacct gttcaccctc accaatcttg gtgcccctgc cgcattcaag tacttcgaca
     4201 ccaccatcga ccggaaacgc tatacctcca ccaaagaagt gctggacgcc accctcatcc
     4261 accagagcat caccggactt tacgaaactc ggattgacct ctcacagctc ggaggggatg
     4321 agggagctgc tagtgatcca aaaaagaaga gaaaggtaga tccaaaaaag aagagaaagg
     4381 tagatccaaa aaagaagaga aaggtagcta gtgacgcgct ggacgatttc gatctcgaca
     4441 tgctgggttc tgatgccctc gatgactttg acctggatat gttgggaagc gacgcattgg
     4501 atgactttga tctggacatg ctcggctccg atgctctgga cgatttcgat ctcgatatgt
     4561 taattaacgc tagcggatcc accggtgtct agaaagcttt gaggccggcc tgcagggatc
     4621 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     4681 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     4741 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     4801 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     4861 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     4921 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     4981 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     5041 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     5101 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     5161 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     5221 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     5281 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     5341 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     5401 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     5461 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     5521 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     5581 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     5641 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     5701 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     5761 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     5821 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     5881 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     5941 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     6001 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     6061 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     6121 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     6181 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     6241 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     6301 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     6361 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     6421 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     6481 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     6541 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     6601 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     6661 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     6721 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     6781 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     6841 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     6901 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     6961 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     7021 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     7081 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     7141 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     7201 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     7261 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     7321 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     7381 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     7441 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     7501 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     7561 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     7621 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     7681 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     7741 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     7801 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     7861 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     7921 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     7981 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     8041 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     8101 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     8161 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     8221 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     8281 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     8341 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     8401 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     8461 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     8521 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     8581 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     8641 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     8701 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     8761 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     8821 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     8881 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     8941 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     9001 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc cacgcgtggt
     9061 accctcgagt tgacattgat tattgagtag ttattaatag taatcaatta cggggtcatt
     9121 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     9181 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     9241 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     9301 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     9361 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     9421 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
     9481 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
//
