LOCUS       sequence-233140-       12869 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentivirus delivery for stable expression of AsCas12a-VPR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12869)
  AUTHORS   Breinig M, Schweitzer AY, Herianto AM, Revia S, Schaefer L, Wendler 
            L, Cobos Galvez A, Tschaharganeh DF
  TITLE     Multiplexed orthogonal genome editing and transcriptional activation
            by Cas12a.
  JOURNAL   Nat Methods. 2019 Jan;16(1):51-54. doi: 10.1038/s41592-018-0262-1. 
            Epub 2018 Dec 17.
  PUBMED    30559432
REFERENCE   2  (bases 1 to 12869)
  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"; doi: 
            "10.1038/s41592-018-0262-1"; journalName: "Nat Methods"; date: 
            "2019-01"; volume: "16"; issue: "1"; pages: "51-54"
COMMENT     Sequence Label: pMB22
FEATURES             Location/Qualifiers
     source          1..12869
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(46..65)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        237..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        618..816
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     771..791
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             834..1014
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1061..1186
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1685..1918
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2103..2147
                     /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    2445..2562
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2680..2891
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2934..2943
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2946..2966
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2991..6908
                     /codon_start=1
                     /product="CRISPR-associated protein from Acidaminococcus 
                     sp. BV3L6 (Zetsche et al., 2015)"
                     /label=AsCpf1
                     /translation="TQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARNDH
                     YKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRNA
                     IHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFDK
                     FTTYFSGFYENRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHFE
                     NVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVLN
                     LAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLLR
                     NENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTGK
                     ITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTLK
                     KQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKAR
                     NYATKKPYSVEKFKLNFQMPTLASGWDVNKEKNNGAILFVKNGLYYLGIMPKQKGRYKA
                     LSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEPL
                     EITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSIDL
                     SSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKGH
                     HGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLKD
                     QKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFFH
                     VPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVIDSTGKILEQRS
                     LNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAVV
                     VLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQF
                     TSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYDV
                     KTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIENH
                     RFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQMR
                     NSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKESK
                     DLKLQNGISNQDWLAYIQELRN"
     regulatory      6628..6637
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6909..6956
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6990..7139
                     /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"
     CDS             7164..7184
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7614..7970
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALL"
     CDS             7989..8558
                     /codon_start=1
                     /product="transcriptional activation domain from the human 
                     herpesvirus 4 (Epstein-Barr virus) replication and 
                     transcription activator Rta/BRLF1 (Hardwick et al., 1992; 
                     Chavez et al., 2015)"
                     /label=Rta AD
                     /translation="RDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWAN
                     RPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKAL
                     REMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEIL
                     DTFLNDECLLHAMHISTGLSIFDTSLF"
     promoter        8586..9085
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(8621..8644)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     9018..9037
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             9106..9705
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(9106..9125)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     9602..9622
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    9758..10346
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9811..9831)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10229..10240)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(10349..10365)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10350..10366)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             10875..11055
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(11117..11705)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11197..11216)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11876..12667,12668..12736))
                     /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     12499..12518
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12737..12841)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
       61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt actgggtctc
      841 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      901 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      961 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
     1021 cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg
     1081 gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat
     1141 tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg
     1201 gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa
     1261 attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct
     1321 gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac
     1381 aggatcagaa gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca
     1441 aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa
     1501 aagtaagacc accgcacagc aagcggccgg ccgcgctgat cttcagacct ggaggaggag
     1561 atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat
     1621 taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg
     1681 gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt
     1741 caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca
     1801 atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca
     1861 agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg
     1921 ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt
     1981 ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag
     2041 aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag
     2101 aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta
     2161 acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag
     2221 gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac
     2281 cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag
     2341 aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac ggatcggcac
     2401 tgcgtgcgcc aattctgcag acaaatggca gtattcatcc acaattttaa aagaaaaggg
     2461 gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa
     2521 actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac
     2581 agcagagatc cagtttggtt agtaccgggc ccgggtcgac tagtgatcag tgtgagggag
     2641 tgtaaagctg gttttcgagt ggctccggtg cccgtcagtg ggcagagcgc acatcgccca
     2701 cagtccccga gaagttgggg ggaggggtcg gcaattgaac cggtgcctag agaaggtggc
     2761 gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg
     2821 gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg
     2881 ccagaacaca ggtgtcgtga cgcggctaac tcgaacgcta gctgtgcgat cgtgccacca
     2941 tggccccaaa gaagaagcgg aaggtcggta tccacggagt cccagcagcc acacagttcg
     3001 agggctttac caacctgtat caggtgagca agacactgcg gtttgagctg atcccacagg
     3061 gcaagaccct gaagcacatc caggagcagg gcttcatcga ggaggacaag gcccgcaatg
     3121 atcactacaa ggagctgaag cccatcatcg atcggatcta caagacctat gccgaccagt
     3181 gcctgcagct ggtgcagctg gattgggaga acctgagcgc cgccatcgac tcctatagaa
     3241 aggagaaaac cgaggagaca aggaacgccc tgatcgagga gcaggccaca tatcgcaatg
     3301 ccatccacga ctacttcatc ggccggacag acaacctgac cgatgccatc aataagagac
     3361 acgccgagat ctacaagggc ctgttcaagg ccgagctgtt taatggcaag gtgctgaagc
     3421 agctgggcac cgtgaccaca accgagcacg agaacgccct gctgcggagc ttcgacaagt
     3481 ttacaaccta cttctccggc ttttatgaga acaggaagaa cgtgttcagc gccgaggata
     3541 tcagcacagc catcccacac cgcatcgtgc aggacaactt ccccaagttt aaggagaatt
     3601 gtcacatctt cacacgcctg atcaccgccg tgcccagcct gcgggagcac tttgagaacg
     3661 tgaagaaggc catcggcatc ttcgtgagca cctccatcga ggaggtgttt tccttccctt
     3721 tttataacca gctgctgaca cagacccaga tcgacctgta taaccagctg ctgggaggaa
     3781 tctctcggga ggcaggcacc gagaagatca agggcctgaa cgaggtgctg aatctggcca
     3841 tccagaagaa tgatgagaca gcccacatca tcgcctccct gccacacaga ttcatccccc
     3901 tgtttaagca gatcctgtcc gataggaaca ccctgtcttt catcctggag gagtttaaga
     3961 gcgacgagga agtgatccag tccttctgca agtacaagac actgctgaga aacgagaacg
     4021 tgctggagac agccgaggcc ctgtttaacg agctgaacag catcgacctg acacacatct
     4081 tcatcagcca caagaagctg gagacaatca gcagcgccct gtgcgaccac tgggatacac
     4141 tgaggaatgc cctgtatgag cggagaatct ccgagctgac aggcaagatc accaagtctg
     4201 ccaaggagaa ggtgcagcgc agcctgaagc acgaggatat caacctgcag gagatcatct
     4261 ctgccgcagg caaggagctg agcgaggcct tcaagcagaa aaccagcgag atcctgtccc
     4321 acgcacacgc cgccctggat cagccactgc ctacaaccct gaagaagcag gaggagaagg
     4381 agatcctgaa gtctcagctg gacagcctgc tgggcctgta ccacctgctg gactggtttg
     4441 ccgtggatga gtccaacgag gtggaccccg agttctctgc ccggctgacc ggcatcaagc
     4501 tggagatgga gccttctctg agcttctaca acaaggccag aaattatgcc accaagaagc
     4561 cctactccgt ggagaagttc aagctgaact ttcagatgcc tacactggcc tctggctggg
     4621 acgtgaataa ggagaagaac aatggcgcca tcctgtttgt gaagaacggc ctgtactatc
     4681 tgggcatcat gccaaagcag aagggcaggt ataaggccct gagcttcgag cccacagaga
     4741 aaaccagcga gggctttgat aagatgtact atgactactt ccctgatgcc gccaagatga
     4801 tcccaaagtg cagcacccag ctgaaggccg tgacagccca ctttcagacc cacacaaccc
     4861 ccatcctgct gtccaacaat ttcatcgagc ctctggagat cacaaaggag atctacgacc
     4921 tgaacaatcc tgagaaggag ccaaagaagt ttcagacagc ctacgccaag aaaaccggcg
     4981 accagaaggg ctacagagag gccctgtgca agtggatcga cttcacaagg gattttctgt
     5041 ccaagtatac caagacaacc tctatcgatc tgtctagcct gcggccatcc tctcagtata
     5101 aggacctggg cgagtactat gccgagctga atcccctgct gtaccacatc agcttccaga
     5161 gaatcgccga gaaggagatc atggatgccg tggagacagg caagctgtac ctgttccaga
     5221 tctataacaa ggactttgcc aagggccacc acggcaagcc taatctgcac acactgtatt
     5281 ggaccggcct gttttctcca gagaacctgg ccaagacaag catcaagctg aatggccagg
     5341 ccgagctgtt ctaccgccct aagtccagga tgaagaggat ggcacaccgg ctgggagaga
     5401 agatgctgaa caagaagctg aaggatcaga aaaccccaat ccccgacacc ctgtaccagg
     5461 agctgtacga ctatgtgaat cacagactgt cccacgacct gtctgatgag gccagggccc
     5521 tgctgcccaa cgtgatcacc aaggaggtgt ctcacgagat catcaaggat aggcgcttta
     5581 ccagcgacaa gttctttttc cacgtgccta tcacactgaa ctatcaggcc gccaattccc
     5641 catctaagtt caaccagagg gtgaatgcct acctgaagga gcaccccgag acacctatca
     5701 tcggcatcga tcggggcgag agaaacctga tctatatcac agtgatcgac tccaccggca
     5761 agatcctgga gcagcggagc ctgaacacca tccagcagtt tgattaccag aagaagctgg
     5821 acaacaggga gaaggagagg gtggcagcaa ggcaggcctg gtctgtggtg ggcacaatca
     5881 aggatctgaa gcagggctat ctgagccagg tcatccacga gatcgtggac ctgatgatcc
     5941 actaccaggc cgtggtggtg ctggagaacc tgaatttcgg ctttaagagc aagaggaccg
     6001 gcatcgccga gaaggccgtg taccagcagt tcgagaagat gctgatcgat aagctgaatt
     6061 gcctggtgct gaaggactat ccagcagaga aagtgggagg cgtgctgaac ccataccagc
     6121 tgacagacca gttcacctcc tttgccaaga tgggcaccca gtctggcttc ctgttttacg
     6181 tgcctgcccc atatacatct aagatcgatc ccctgaccgg cttcgtggac cccttcgtgt
     6241 ggaaaaccat caagaatcac gagagccgca agcacttcct ggagggcttc gactttctgc
     6301 actacgacgt gaaaaccggc gacttcatcc tgcactttaa gatgaacaga aatctgtcct
     6361 tccagagggg cctgcccggc tttatgcctg catgggatat cgtgttcgag aagaacgaga
     6421 cacagtttga cgccaagggc acccctttca tcgccggcaa gagaatcgtg ccagtgatcg
     6481 agaatcacag attcaccggc agataccggg acctgtatcc tgccaacgag ctgatcgccc
     6541 tgctggagga gaagggcatc gtgttcaggg atggctccaa catcctgcca aagctgctgg
     6601 agaatgacga ttctcacgcc atcgacacca tggtggccct gatccgcagc gtgctgcaga
     6661 tgcggaactc caatgccgcc acaggcgagg actatatcaa cagccccgtg cgcgatctga
     6721 atggcgtgtg cttcgactcc cggtttcaga acccagagtg gcccatggac gccgatgcca
     6781 atggcgccta ccacatcgcc ctgaagggcc agctgctgct gaatcacctg aaggagagca
     6841 aggatctgaa gctgcagaac ggcatctcca atcaggactg gctggcctac atccaggagc
     6901 tgcgcaacaa aaggccggcg gccacgaaaa aggccggcca ggcaaaaaag aaaaagggat
     6961 ccaaagaggc cagcggttcc ggacgggctg acgcattgga cgattttgat ctggatatgc
     7021 tgggaagtga cgccctcgat gattttgacc ttgacatgct tggttcggat gcccttgatg
     7081 actttgacct cgacatgctc ggcagtgacg cccttgatga tttcgacctg gacatgctga
     7141 ttaactctag aagttccgga tctccgaaaa agaaacgcaa agttggtagc cagtacctgc
     7201 ccgacaccga cgaccggcac cggatcgagg aaaagcggaa gcggacctac gagacattca
     7261 agagcatcat gaagaagtcc cccttcagcg gccccaccga ccctagacct ccacctagaa
     7321 gaatcgccgt gcccagcaga tccagcgcca gcgtgccaaa acctgccccc cagccttacc
     7381 ccttcaccag cagcctgagc accatcaact acgacgagtt ccctaccatg gtgttcccca
     7441 gcggccagat ctctcaggcc tctgctctgg ctccagcccc tcctcaggtg ctgcctcagg
     7501 ctcctgctcc tgcaccagct ccagccatgg tgtctgcact ggctcaggca ccagcacccg
     7561 tgcctgtgct ggctcctgga cctccacagg ctgtggctcc accagcccct aaacctacac
     7621 aggccggcga gggcacactg tctgaagctc tgctgcagct gcagttcgac gacgaggatc
     7681 tgggagccct gctgggaaac agcaccgatc ctgccgtgtt caccgacctg gccagcgtgg
     7741 acaacagcga gttccagcag ctgctgaacc agggcatccc tgtggcccct cacaccaccg
     7801 agcccatgct gatggaatac cccgaggcca tcacccggct cgtgacaggc gctcagaggc
     7861 ctcctgatcc agctcctgcc cctctgggag caccaggcct gcctaatgga ctgctgtctg
     7921 gcgacgagga cttcagctct atcgccgata tggatttctc agccttgctg ggctctggca
     7981 gcggcagccg ggattccagg gaagggatgt ttttgccgaa gcctgaggcc ggctccgcta
     8041 ttagtgacgt gtttgagggc cgcgaggtgt gccagccaaa acgaatccgg ccatttcatc
     8101 ctccaggaag tccatgggcc aaccgcccac tccccgccag cctcgcacca acaccaaccg
     8161 gtccagtaca tgagccagtc gggtcactga ccccggcacc agtccctcag ccactggatc
     8221 cagcgcccgc agtgactccc gaggccagtc acctgttgga ggatcccgat gaagagacga
     8281 gccaggctgt caaagccctt cgggagatgg ccgatactgt gattccccag aaggaagagg
     8341 ctgcaatctg tggccaaatg gacctttccc atccgccccc aaggggccat ctggatgagc
     8401 tgacaaccac acttgagtcc atgaccgagg atctgaacct ggactcaccc ctgaccccgg
     8461 aattgaacga gattctggat accttcctga acgacgagtg cctcttgcat gccatgcata
     8521 tcagcacagg actgtccatc ttcgacacat ctctgttttg acctaggaat cccgaggcct
     8581 ctaccgggta ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt tagcagcccc
     8641 gctgggcact tggcgctaca caagtggcct ctggcctcgc acacattcca catccaccgg
     8701 taggcgccaa ccggctccgt tctttggtgg ccccttcgcg ccaccttcta ctcctcccct
     8761 agtcaggaag ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac aaatggaagt
     8821 agcacgtctc actagtctcg tgcagatgga cagcaccgct gagcaatgga agcgggtagg
     8881 cctttggggc agcggccaat agcagctttg ctccttcgct ttctgggctc agaggctggg
     8941 aaggggtggg tccgggggcg ggctcagggg cgggctcagg ggcggggcgg gcgcccgaag
     9001 gtcctccgga ggcccggcat tctgcacgct tcaaaagcgc acgtctgccg cgctgttctc
     9061 ctcttcctca tctccgggcc tttcgacctg cagcccaagc ttaccatgac cgagtacaag
     9121 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc
     9181 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
     9241 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     9301 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     9361 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     9421 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     9481 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     9541 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     9601 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     9661 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgaaaacg ttgttgtttg
     9721 gggttgaatt actcttctag accgcggcgc gcccgataat caacctctgg attacaaaat
     9781 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     9841 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     9901 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     9961 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
    10021 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
    10081 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
    10141 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
    10201 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
    10261 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
    10321 gatctccctt tgggccgcct ccccgcatcg ataccgtcga cctcgatcga gacctagaaa
    10381 aacatggagc aatcacaagt agcaatacag cagctaccaa tgctgattgt gcctggctag
    10441 aagcacaaga ggaggaggag gtgggttttc cagtcacacc tcaggtacct ttaagaccaa
    10501 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
    10561 ggctaattca ctcccaacga agacaagata tccttgatct gtggatctac cacacacaag
    10621 gctacttccc tgattggcag aactacacac cagggccagg gatcagatat ccactgacct
    10681 ttggatggtg ctacaagcta gtaccagttg agcaagagaa ggtagaagaa gccaatgaag
    10741 gagagaacac ccgcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag
    10801 aagtattaga gtggaggttt gacagccgcc tagcatttca tcacatggcc cgagagctgc
    10861 atccggactg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct
    10921 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
    10981 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
    11041 ggaaaatctc tagcagcatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
    11101 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
    11161 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
    11221 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
    11281 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
    11341 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
    11401 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
    11461 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
    11521 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
    11581 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
    11641 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
    11701 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
    11761 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
    11821 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
    11881 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
    11941 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
    12001 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
    12061 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
    12121 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
    12181 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
    12241 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
    12301 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
    12361 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
    12421 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
    12481 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
    12541 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
    12601 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
    12661 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
    12721 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
    12781 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
    12841 gcacatttcc ccgaaaagtg ccacctgac
//
