LOCUS       sequence-308054-       11432 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector expressing human codon-optimized _C-terminal 
            Myc-tagged _Acidaminococcus sp. BV3L6 (As)-Cas12a nuclease with N- 
            and C-terminal NLS and Neomycin/G418/Geneticin resistance from 
            EF-1alpha promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11432)
  AUTHORS   Gonatopoulos-Pournatzis T, Aregger M, Brown KR, Farhangmehr S, 
            Braunschweig U, Ward HN, Ha KCH, Weiss A, Billmann M, Durbic T, 
            Myers CL, Blencowe BJ, Moffat J
  TITLE     Genetic interaction mapping and exon-resolution functional genomics 
            with a hybrid Cas9-Cas12a platform.
  JOURNAL   Nat Biotechnol. 2020 May;38(5):638-648. doi: 
            10.1038/s41587-020-0437-z. Epub 2020 Mar 16.
  PUBMED    32249828
REFERENCE   2  (bases 1 to 11432)
  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/s41587-020-0437-z"; journalName: "Nat Biotechnol"; date: 
            "2020-05"; volume: "38"; issue: "5"; pages: "638-648"
COMMENT     Sequence Label: plenti-As-Cas12a-2xNLS
FEATURES             Location/Qualifiers
     source          1..11432
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    88..109
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(124..129,130..147,148..154)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    162..178
                     /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)."
     primer_bind     167..189
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     186..202
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     186..202
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     221..241
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        223..241
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        269..495
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             496..676
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    723..848
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1341..1574
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1759..1803
                     /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"
     primer_bind     1981..2001
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     misc_feature    2202..2319
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2388..2599
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             2627..2647
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2672..6589
                     /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      6309..6318
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6590..6637
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6644..6673
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             6683..6736
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             6740..7534
                     /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
                     GLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6794..6813)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7404..7423
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     misc_feature    7553..8141
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7606..7626)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8024..8035)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8213..8446
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8524..8645
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8561..8580)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8615..8634
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8685..8820
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8840..8859)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8841..8859)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8869..8886)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8869..8885)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8878..8900)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9027..9482
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9114..9133)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9324..9345
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9508..9612
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9613..9681,9682..10473)
                     /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(9831..9850)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10644..11232
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11133..11152
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11386..11403
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
       61 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
      121 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
      181 tcacacagga aacagctatg accatgatta cgccaagcgc gcaattaacc ctcactaaag
      241 ggaacaaaag ctggagctgc aagcttaatg tagtcttatg caatactctt gtagtcttgc
      301 aacatggtaa cgatgagtta gcaacatgcc ttacaaggag agaaaaagca ccgtgcatgc
      361 cgattggtgg aagtaaggtg gtacgatcgt gccttattag gaaggcaaca gacgggtctg
      421 acatggattg gacgaaccac tgaattgccg cattgcagag atattgtatt taagtgccta
      481 gctcgataca taaacgggtc tctctggtta gaccagatct gagcctggga gctctctggc
      541 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
      601 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
      661 tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg
      721 agctctctcg acgcaggact cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc
      781 gactggtgag tacgccaaaa attttgacta gcggaggcta gaaggagaga gatgggtgcg
      841 agagcgtcag tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc
      901 cagggggaaa gaaaaaatat aaattaaaac atatagtatg ggcaagcagg gagctagaac
      961 gattcgcagt taatcctggc ctgttagaaa catcagaagg ctgtagacaa atactgggac
     1021 agctacaacc atcccttcag acaggatcag aagaacttag atcattatat aatacagtag
     1081 caaccctcta ttgtgtgcat caaaggatag agataaaaga caccaaggaa gctttagaca
     1141 agatagagga agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc
     1201 agacctggag gaggagatat gagggacaat tggagaagtg aattatataa atataaagta
     1261 gtaaaaattg aaccattagg agtagcaccc accaaggcaa agagaagagt ggtgcagaga
     1321 gaaaaaagag cagtgggaat aggagctttg ttccttgggt tcttgggagc agcaggaagc
     1381 actatgggcg cagcgtcaat gacgctgacg gtacaggcca gacaattatt gtctggtata
     1441 gtgcagcagc agaacaattt gctgagggct attgaggcgc aacagcatct gttgcaactc
     1501 acagtctggg gcatcaagca gctccaggca agaatcctgg ctgtggaaag atacctaaag
     1561 gatcaacagc tcctggggat ttggggttgc tctggaaaac tcatttgcac cactgctgtg
     1621 ccttggaatg ctagttggag taataaatct ctggaacaga tttggaatca cacgacctgg
     1681 atggagtggg acagagaaat taacaattac acaagcttaa tacactcctt aattgaagaa
     1741 tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg aattagataa atgggcaagt
     1801 ttgtggaatt ggtttaacat aacaaattgg ctgtggtata taaaattatt cataatgata
     1861 gtaggaggct tggtaggttt aagaatagtt tttgctgtac tttctatagt gaatagagtt
     1921 aggcagggat attcaccatt atcgtttcag acccacctcc caaccccgag gggacccaga
     1981 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
     2041 ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
     2101 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
     2161 aaattctaga tcttgagaca aatggcagta ttcatccaca attttaaaag aaaagggggg
     2221 attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga catacaaact
     2281 aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta cagggacagc
     2341 agagatccac tttggcgccg gctcgagtgg ctccggtgcc cgtcagtggg cagagcgcac
     2401 atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgaaccg gtgcctagag
     2461 aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga
     2521 gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg
     2581 gtttgccgcc agaacacagg tgtcgtgacg cggttacacc atggccccaa agaagaagcg
     2641 gaaggtcggt atccacggag tcccagcagc cacacagttc gagggcttta ccaacctgta
     2701 tcaggtgagc aagacactgc ggtttgagct gatcccacag ggcaagaccc tgaagcacat
     2761 ccaggagcag ggcttcatcg aggaggacaa ggcccgcaat gatcactaca aggagctgaa
     2821 gcccatcatc gatcggatct acaagaccta tgccgaccag tgcctgcagc tggtgcagct
     2881 ggattgggag aacctgagcg ccgccatcga ctcctataga aaggagaaaa ccgaggagac
     2941 aaggaacgcc ctgatcgagg agcaggccac atatcgcaat gccatccacg actacttcat
     3001 cggccggaca gacaacctga ccgatgccat caataagaga cacgccgaga tctacaaggg
     3061 cctgttcaag gccgagctgt ttaatggcaa ggtgctgaag cagctgggca ccgtgaccac
     3121 aaccgagcac gagaacgccc tgctgcggag cttcgacaag tttacaacct acttctccgg
     3181 cttttatgag aacaggaaga acgtgttcag cgccgaggat atcagcacag ccatcccaca
     3241 ccgcatcgtg caggacaact tccccaagtt taaggagaat tgtcacatct tcacacgcct
     3301 gatcaccgcc gtgcccagcc tgcgggagca ctttgagaac gtgaagaagg ccatcggcat
     3361 cttcgtgagc acctccatcg aggaggtgtt ttccttccct ttttataacc agctgctgac
     3421 acagacccag atcgacctgt ataaccagct gctgggagga atctctcggg aggcaggcac
     3481 cgagaagatc aagggcctga acgaggtgct gaatctggcc atccagaaga atgatgagac
     3541 agcccacatc atcgcctccc tgccacacag attcatcccc ctgtttaagc agatcctgtc
     3601 cgataggaac accctgtctt tcatcctgga ggagtttaag agcgacgagg aagtgatcca
     3661 gtccttctgc aagtacaaga cactgctgag aaacgagaac gtgctggaga cagccgaggc
     3721 cctgtttaac gagctgaaca gcatcgacct gacacacatc ttcatcagcc acaagaagct
     3781 ggagacaatc agcagcgccc tgtgcgacca ctgggataca ctgaggaatg ccctgtatga
     3841 gcggagaatc tccgagctga caggcaagat caccaagtct gccaaggaga aggtgcagcg
     3901 cagcctgaag cacgaggata tcaacctgca ggagatcatc tctgccgcag gcaaggagct
     3961 gagcgaggcc ttcaagcaga aaaccagcga gatcctgtcc cacgcacacg ccgccctgga
     4021 tcagccactg cctacaaccc tgaagaagca ggaggagaag gagatcctga agtctcagct
     4081 ggacagcctg ctgggcctgt accacctgct ggactggttt gccgtggatg agtccaacga
     4141 ggtggacccc gagttctctg cccggctgac cggcatcaag ctggagatgg agccttctct
     4201 gagcttctac aacaaggcca gaaattatgc caccaagaag ccctactccg tggagaagtt
     4261 caagctgaac tttcagatgc ctacactggc ctctggctgg gacgtgaata aggagaagaa
     4321 caatggcgcc atcctgtttg tgaagaacgg cctgtactat ctgggcatca tgccaaagca
     4381 gaagggcagg tataaggccc tgagcttcga gcccacagag aaaaccagcg agggctttga
     4441 taagatgtac tatgactact tccctgatgc cgccaagatg atcccaaagt gcagcaccca
     4501 gctgaaggcc gtgacagccc actttcagac ccacacaacc cccatcctgc tgtccaacaa
     4561 tttcatcgag cctctggaga tcacaaagga gatctacgac ctgaacaatc ctgagaagga
     4621 gccaaagaag tttcagacag cctacgccaa gaaaaccggc gaccagaagg gctacagaga
     4681 ggccctgtgc aagtggatcg acttcacaag ggattttctg tccaagtata ccaagacaac
     4741 ctctatcgat ctgtctagcc tgcggccatc ctctcagtat aaggacctgg gcgagtacta
     4801 tgccgagctg aatcccctgc tgtaccacat cagcttccag agaatcgccg agaaggagat
     4861 catggatgcc gtggagacag gcaagctgta cctgttccag atctataaca aggactttgc
     4921 caagggccac cacggcaagc ctaatctgca cacactgtat tggaccggcc tgttttctcc
     4981 agagaacctg gccaagacaa gcatcaagct gaatggccag gccgagctgt tctaccgccc
     5041 taagtccagg atgaagagga tggcacaccg gctgggagag aagatgctga acaagaagct
     5101 gaaggatcag aaaaccccaa tccccgacac cctgtaccag gagctgtacg actatgtgaa
     5161 tcacagactg tcccacgacc tgtctgatga ggccagggcc ctgctgccca acgtgatcac
     5221 caaggaggtg tctcacgaga tcatcaagga taggcgcttt accagcgaca agttcttttt
     5281 ccacgtgcct atcacactga actatcaggc cgccaattcc ccatctaagt tcaaccagag
     5341 ggtgaatgcc tacctgaagg agcaccccga gacacctatc atcggcatcg atcggggcga
     5401 gagaaacctg atctatatca cagtgatcga ctccaccggc aagatcctgg agcagcggag
     5461 cctgaacacc atccagcagt ttgattacca gaagaagctg gacaacaggg agaaggagag
     5521 ggtggcagca aggcaggcct ggtctgtggt gggcacaatc aaggatctga agcagggcta
     5581 tctgagccag gtcatccacg agatcgtgga cctgatgatc cactaccagg ccgtggtggt
     5641 gctggagaac ctgaatttcg gctttaagag caagaggacc ggcatcgccg agaaggccgt
     5701 gtaccagcag ttcgagaaga tgctgatcga taagctgaat tgcctggtgc tgaaggacta
     5761 tccagcagag aaagtgggag gcgtgctgaa cccataccag ctgacagacc agttcacctc
     5821 ctttgccaag atgggcaccc agtctggctt cctgttttac gtgcctgccc catatacatc
     5881 taagatcgat cccctgaccg gcttcgtgga ccccttcgtg tggaaaacca tcaagaatca
     5941 cgagagccgc aagcacttcc tggagggctt cgactttctg cactacgacg tgaaaaccgg
     6001 cgacttcatc ctgcacttta agatgaacag aaatctgtcc ttccagaggg gcctgcccgg
     6061 ctttatgcct gcatgggata tcgtgttcga gaagaacgag acacagtttg acgccaaggg
     6121 cacccctttc atcgccggca agagaatcgt gccagtgatc gagaatcaca gattcaccgg
     6181 cagataccgg gacctgtatc ctgccaacga gctgatcgcc ctgctggagg agaagggcat
     6241 cgtgttcagg gatggctcca acatcctgcc aaagctgctg gagaatgacg attctcacgc
     6301 catcgacacc atggtggccc tgatccgcag cgtgctgcag atgcggaact ccaatgccgc
     6361 cacaggcgag gactatatca acagccccgt gcgcgatctg aatggcgtgt gcttcgactc
     6421 ccggtttcag aacccagagt ggcccatgga cgccgatgcc aatggcgcct accacatcgc
     6481 cctgaagggc cagctgctgc tgaatcacct gaaggagagc aaggatctga agctgcagaa
     6541 cggcatctcc aatcaggact ggctggccta catccaggag ctgcgcaaca aaaggccggc
     6601 ggccacgaaa aaggccggcc aggcaaaaaa gaaaaaggga tccgaacaaa aactcatctc
     6661 agaagaggat ctgggcagtg gagagggcag aggaagtctg ctaacatgcg gtgacgtcga
     6721 ggagaatcct ggcccaacca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     6781 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     6841 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     6901 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     6961 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     7021 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     7081 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     7141 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     7201 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     7261 caaggcgcgc atgcccgacg gcgatgatct cgtcgtgacc catggcgatg cctgcttgcc
     7321 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     7381 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     7441 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     7501 cgccttctat cgccttcttg acgagttctt ctgataaacg cgttaagtcg acaatcaacc
     7561 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     7621 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     7681 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     7741 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     7801 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     7861 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     7921 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt
     7981 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     8041 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     8101 ccctcagacg agtcggatct ccctttgggc cgcctccccg cgtcgacttt aagaccaatg
     8161 acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg
     8221 ctaattcact cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta
     8281 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     8341 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     8401 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtac gtatagtagt
     8461 tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag
     8521 aggaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     8581 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     8641 tcttatcatg tctggctcta gctatcccgc ccctaactcc gcccatcccg cccctaactc
     8701 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     8761 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     8821 tagggacgta cccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt
     8881 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     8941 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     9001 gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg
     9061 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     9121 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     9181 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     9241 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     9301 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     9361 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     9421 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     9481 ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     9541 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     9601 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     9661 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     9721 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     9781 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     9841 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     9901 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     9961 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
    10021 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
    10081 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
    10141 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
    10201 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
    10261 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
    10321 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
    10381 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
    10441 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
    10501 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
    10561 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
    10621 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
    10681 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
    10741 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
    10801 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
    10861 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
    10921 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
    10981 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
    11041 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
    11101 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
    11161 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
    11221 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
    11281 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
    11341 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
    11401 gaggaagcgg aagagcgccc aatacgcaaa cc
//
