LOCUS       sequence-413627-       12294 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector expressing human codon-optimized opCas12a-Ultra 
            with 6xNLS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12294)
  AUTHORS   Xiao MS, Damodaran AP, Kumari B, Dickson E, Xing K, On TA, Parab N, 
            King HE, Perez AR, Guiblet WM, Duncan G, Che A, Chari R, Andresson 
            T, Vidigal JA, Weatheritt RJ, Aregger M, Gonatopoulos-Pournatzis T
  TITLE     Genome-scale exon perturbation screens uncover exons critical for 
            cell fitness.
  JOURNAL   Mol Cell. 2024 Jul 11;84(13):2553-2572.e19. doi: 
            10.1016/j.molcel.2024.05.024. Epub 2024 Jun 24.
  PUBMED    38917794
REFERENCE   2  (bases 1 to 12294)
  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: 
            "10.1016/j.molcel.2024.05.024"; journalName: "Mol Cell"; date: 
            "2024-07-11- 11"; volume: "84"; issue: "13"; pages: "2553-2572.e19"
COMMENT     Sequence Label: pLenti-opCas12a-Ultra-6xNLS
FEATURES             Location/Qualifiers
     source          1..12294
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     240..260
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        242..260
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        288..514
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             515..695
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    742..867
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1360..1593
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1778..1822
                     /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    2006..2123
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2172..3349
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2403..3340
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3297..3317
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      3355..3364
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3364..3387
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3388..7305
                     /codon_start=1
                     /product="CRISPR-associated protein from Acidaminococcus 
                     sp. BV3L6 (Zetsche et al., 2015)"
                     /label=AsCpf1
                     /translation="TQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARNDH
                     YKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRNA
                     IHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFDK
                     FTTYFSGFYRNRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHFE
                     NVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVLN
                     LAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLLR
                     NENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTGK
                     ITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTLK
                     KQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKAR
                     NYATKKPYSVEKFKLNFQRPTLARGWDVNKEKNNGAILFVKNGLYYLGIMPKQKGRYKA
                     LSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEPL
                     EITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSIDL
                     SSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKGH
                     HGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLKD
                     QKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFLFH
                     VPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVIDSTGKILEQRS
                     LNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAVV
                     VLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQF
                     TSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYDV
                     KTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIENH
                     RFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQMR
                     NSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKESK
                     DLKLQNGISNQDWLAYIQELRN"
     regulatory      7025..7034
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7315..7341
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             7351..7377
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             7387..7413
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             7423..7449
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             7459..7485
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             7495..7521
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             7528..7557
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             7567..7620
                     /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             7624..8418
                     /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(7678..7697)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8288..8307
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     misc_feature    8434..9022
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8487..8507)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8905..8916)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9094..9327
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    9405..9526
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9442..9461)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9496..9515
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      9566..9701
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(9721..9740)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9722..9740)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9750..9767)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9750..9766)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9759..9781)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9908..10363
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9995..10014)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10205..10226
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        10389..10493
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10494..10562,10563..11354)
                     /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(10712..10731)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11525..12113
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12014..12033
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12267..12284
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg
      241 caattaaccc tcactaaagg gaacaaaagc tggagctgca agcttaatgt agtcttatgc
      301 aatactcttg tagtcttgca acatggtaac gatgagttag caacatgcct tacaaggaga
      361 gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg ccttattagg
      421 aaggcaacag acgggtctga catggattgg acgaaccact gaattgccgc attgcagaga
      481 tattgtattt aagtgcctag ctcgatacat aaacgggtct ctctggttag accagatctg
      541 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
      601 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
      661 cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa
      721 gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg
      781 gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag
      841 aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg
      901 gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg
      961 gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc
     1021 tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga
     1081 tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac
     1141 accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag
     1201 caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga
     1261 attatataaa tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa
     1321 gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt
     1381 cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag
     1441 acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca
     1501 acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc
     1561 tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact
     1621 catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat
     1681 ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat
     1741 acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga
     1801 attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat
     1861 aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact
     1921 ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc
     1981 aaccccgagg ggacccagac cacaatttta aaagaaaagg ggggattggg gggtacagtg
     2041 caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa ttacaaaaac
     2101 aaattacaaa aattcaaaat tttcgggttt attacaggga cagcagagat ccactttggc
     2161 gccggctcga gggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg
     2221 agaagttggg gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa
     2281 actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt
     2341 atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac
     2401 aggtaagtgc cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg
     2461 tgccttgaat tacttccact ggctgcagta cgtgattctt gatcccgagc ttcgggttgg
     2521 aagtgggtgg gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt
     2581 tgaggcctgg cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg
     2641 tctcgctgct ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct
     2701 ttttttctgg caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt
     2761 ttttggggcc gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg
     2821 gggcctgcga gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc
     2881 tctggtgcct ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg
     2941 gtcggcacca gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc
     3001 aaaatggagg acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag
     3061 ggcctttccg tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag
     3121 gcacctcgat tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt
     3181 ttatgcgatg gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca
     3241 cttgatgtaa ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa
     3301 gcctcagaca gtggttcaaa gtttttttct tccatttcag gtgtcgtgag atccgccacc
     3361 atggattaca aagacgatga cgataagaca cagttcgagg gctttaccaa cctgtatcag
     3421 gtgagcaaga cactgcggtt tgagctgatc ccacagggca agaccctgaa gcacatccag
     3481 gagcagggct tcatcgagga ggacaaggcc cgcaatgatc actacaagga gctgaagccc
     3541 atcatcgatc ggatctacaa gacctatgcc gaccagtgcc tgcagctggt gcagctggat
     3601 tgggagaacc tgagcgccgc catcgactcc tatagaaagg agaaaaccga ggagacaagg
     3661 aacgccctga tcgaggagca ggccacatat cgcaatgcca tccacgacta cttcatcggc
     3721 cggacagaca acctgaccga tgccatcaat aagagacacg ccgagatcta caagggcctg
     3781 ttcaaggccg agctgtttaa tggcaaggtg ctgaagcagc tgggcaccgt gaccacaacc
     3841 gagcacgaga acgccctgct gcggagcttc gacaagttta caacctactt ctccggcttt
     3901 tatcggaaca ggaagaacgt gttcagcgcc gaggatatca gcacagccat cccacaccgc
     3961 atcgtgcagg acaacttccc caagtttaag gagaattgtc acatcttcac acgcctgatc
     4021 accgccgtgc ccagcctgcg ggagcacttt gagaacgtga agaaggccat cggcatcttc
     4081 gtgagcacct ccatcgagga ggtgttttcc ttcccttttt ataaccagct gctgacacag
     4141 acccagatcg acctgtataa ccagctgctg ggaggaatct ctcgggaggc aggcaccgag
     4201 aagatcaagg gcctgaacga ggtgctgaat ctggccatcc agaagaatga tgagacagcc
     4261 cacatcatcg cctccctgcc acacagattc atccccctgt ttaagcagat cctgtccgat
     4321 aggaacaccc tgtctttcat cctggaggag tttaagagcg acgaggaagt gatccagtcc
     4381 ttctgcaagt acaagacact gctgagaaac gagaacgtgc tggagacagc cgaggccctg
     4441 tttaacgagc tgaacagcat cgacctgaca cacatcttca tcagccacaa gaagctggag
     4501 acaatcagca gcgccctgtg cgaccactgg gatacactga ggaatgccct gtatgagcgg
     4561 agaatctccg agctgacagg caagatcacc aagtctgcca aggagaaggt gcagcgcagc
     4621 ctgaagcacg aggatatcaa cctgcaggag atcatctctg ccgcaggcaa ggagctgagc
     4681 gaggccttca agcagaaaac cagcgagatc ctgtcccacg cacacgccgc cctggatcag
     4741 ccactgccta caaccctgaa gaagcaggag gagaaggaga tcctgaagtc tcagctggac
     4801 agcctgctgg gcctgtacca cctgctggac tggtttgccg tggatgagtc caacgaggtg
     4861 gaccccgagt tctctgcccg gctgaccggc atcaagctgg agatggagcc ttctctgagc
     4921 ttctacaaca aggccagaaa ttatgccacc aagaagccct actccgtgga gaagttcaag
     4981 ctgaactttc agcggcctac actggcccgt ggctgggacg tgaataagga gaagaacaat
     5041 ggcgccatcc tgtttgtgaa gaacggcctg tactatctgg gcatcatgcc aaagcagaag
     5101 ggcaggtata aggccctgag cttcgagccc acagagaaaa ccagcgaggg ctttgataag
     5161 atgtactatg actacttccc tgatgccgcc aagatgatcc caaagtgcag cacccagctg
     5221 aaggccgtga cagcccactt tcagacccac acaaccccca tcctgctgtc caacaatttc
     5281 atcgagcctc tggagatcac aaaggagatc tacgacctga acaatcctga gaaggagcca
     5341 aagaagtttc agacagccta cgccaagaaa accggcgacc agaagggcta cagagaggcc
     5401 ctgtgcaagt ggatcgactt cacaagggat tttctgtcca agtataccaa gacaacctct
     5461 atcgatctgt ctagcctgcg gccatcctct cagtataagg acctgggcga gtactatgcc
     5521 gagctgaatc ccctgctgta ccacatcagc ttccagagaa tcgccgagaa ggagatcatg
     5581 gatgccgtgg agacaggcaa gctgtacctg ttccagatct ataacaagga ctttgccaag
     5641 ggccaccacg gcaagcctaa tctgcacaca ctgtattgga ccggcctgtt ttctccagag
     5701 aacctggcca agacaagcat caagctgaat ggccaggccg agctgttcta ccgccctaag
     5761 tccaggatga agaggatggc acaccggctg ggagagaaga tgctgaacaa gaagctgaag
     5821 gatcagaaaa ccccaatccc cgacaccctg taccaggagc tgtacgacta tgtgaatcac
     5881 agactgtccc acgacctgtc tgatgaggcc agggccctgc tgcccaacgt gatcaccaag
     5941 gaggtgtctc acgagatcat caaggatagg cgctttacca gcgacaagtt cctgttccac
     6001 gtgcctatca cactgaacta tcaggccgcc aattccccat ctaagttcaa ccagagggtg
     6061 aatgcctacc tgaaggagca ccccgagaca cctatcatcg gcatcgatcg gggcgagaga
     6121 aacctgatct atatcacagt gatcgactcc accggcaaga tcctggagca gcggagcctg
     6181 aacaccatcc agcagtttga ttaccagaag aagctggaca acagggagaa ggagagggtg
     6241 gcagcaaggc aggcctggtc tgtggtgggc acaatcaagg atctgaagca gggctatctg
     6301 agccaggtca tccacgagat cgtggacctg atgatccact accaggccgt ggtggtgctg
     6361 gagaacctga atttcggctt taagagcaag aggaccggca tcgccgagaa ggccgtgtac
     6421 cagcagttcg agaagatgct gatcgataag ctgaattgcc tggtgctgaa ggactatcca
     6481 gcagagaaag tgggaggcgt gctgaaccca taccagctga cagaccagtt cacctccttt
     6541 gccaagatgg gcacccagtc tggcttcctg ttttacgtgc ctgccccata tacatctaag
     6601 atcgatcccc tgaccggctt cgtggacccc ttcgtgtgga aaaccatcaa gaatcacgag
     6661 agccgcaagc acttcctgga gggcttcgac tttctgcact acgacgtgaa aaccggcgac
     6721 ttcatcctgc actttaagat gaacagaaat ctgtccttcc agaggggcct gcccggcttt
     6781 atgcctgcat gggatatcgt gttcgagaag aacgagacac agtttgacgc caagggcacc
     6841 cctttcatcg ccggcaagag aatcgtgcca gtgatcgaga atcacagatt caccggcaga
     6901 taccgggacc tgtatcctgc caacgagctg atcgccctgc tggaggagaa gggcatcgtg
     6961 ttcagggatg gctccaacat cctgccaaag ctgctggaga atgacgattc tcacgccatc
     7021 gacaccatgg tggccctgat ccgcagcgtg ctgcagatgc ggaactccaa tgccgccaca
     7081 ggcgaggact atatcaacag ccccgtgcgc gatctgaatg gcgtgtgctt cgactcccgg
     7141 tttcagaacc cagagtggcc catggacgcc gatgccaatg gcgcctacca catcgccctg
     7201 aagggccagc tgctgctgaa tcacctgaag gagagcaagg atctgaagct gcagaacggc
     7261 atctccaatc aggactggct ggcctacatc caggagctgc gcaacggcgg ctcccctgct
     7321 gctaaacgtg ttaagcttga tgggggtagc ccggcagcca agagagtcaa actcgacggc
     7381 ggctccccag cggcaaaaag ggtgaaacta gacgggggta gccccgccgc gaagcgtgta
     7441 aagctggatg gaggatcgcc tgcggctaag cgagtcaaat tagacggtgg gtcaccggct
     7501 gcaaagcgcg tcaaattgga tgctagcgaa caaaaactca tctcagaaga ggatctgggc
     7561 agtggagagg gcagaggaag tctgctaaca tgcggtgacg tcgaggagaa tcctggccca
     7621 accatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     7681 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     7741 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     7801 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     7861 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     7921 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     7981 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     8041 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     8101 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     8161 gacggcgatg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     8221 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     8281 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     8341 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     8401 cttgacgagt tcttctgaac gcgttaagtc gacaatcaac ctctggatta caaaatttgt
     8461 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     8521 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     8581 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     8641 gtgtgcactg tgtttgctga cgcaaccccc actggttggg acattgccac cacctgtcag
     8701 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     8761 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     8821 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc
     8881 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     8941 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     9001 tccctttggg ccgcctcccc gcgtcgactt taagaccaat gacttacaag gcagctgtag
     9061 atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa
     9121 gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg
     9181 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     9241 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     9301 tttagtcagt gtggaaaatc tctagcagta cgtatagtag ttcatgtcat cttattattc
     9361 agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag
     9421 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     9481 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct
     9541 agctatcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     9601 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     9661 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagggacgt acccaattcg
     9721 ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg
     9781 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     9841 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     9901 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     9961 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
    10021 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
    10081 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
    10141 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
    10201 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
    10261 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
    10321 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg
    10381 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
    10441 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
    10501 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
    10561 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
    10621 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
    10681 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
    10741 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
    10801 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
    10861 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
    10921 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
    10981 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
    11041 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
    11101 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
    11161 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
    11221 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
    11281 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
    11341 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
    11401 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
    11461 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
    11521 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
    11581 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
    11641 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
    11701 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
    11761 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
    11821 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
    11881 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
    11941 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
    12001 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
    12061 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
    12121 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
    12181 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
    12241 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaag
//
