LOCUS       sequence-413586-       12270 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector expressing human codon-optimized UltraCas12a with 
            6xNLS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12270)
  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 12270)
  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-UltraCas12a-6xNLS
FEATURES             Location/Qualifiers
     source          1..12270
                     /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"
     CDS             3361..7281
                     /codon_start=1
                     /product="CRISPR-associated protein from Acidaminococcus 
                     sp. BV3L6 (Zetsche et al., 2015)"
                     /label=AsCpf1
                     /translation="MTQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARND
                     HYKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRN
                     AIHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFD
                     KFTTYFSGFYENRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHF
                     ENVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVL
                     NLAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLL
                     RNENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTG
                     KITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTL
                     KKQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKA
                     RNYATKKPYSVEKFKLNFQRPTLASGWDVNKEKNNGAILFVKNGLYYLGIMPKQKGRYK
                     ALSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEP
                     LEITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSID
                     LSSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKG
                     HHGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLK
                     DQKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFLF
                     HVPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVIDSTGKILEQR
                     SLNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAV
                     VVLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQ
                     FTSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYD
                     VKTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIEN
                     HRFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQM
                     RNSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKES
                     KDLKLQNGISNQDWLAYIQELRN"
     regulatory      7001..7010
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7291..7317
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             7327..7353
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             7363..7389
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             7399..7425
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             7435..7461
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             7471..7497
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             7504..7533
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             7543..7596
                     /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             7600..8394
                     /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(7654..7673)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8264..8283
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     misc_feature    8410..8998
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8463..8483)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8881..8892)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9070..9303
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    9381..9502
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9418..9437)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9472..9491
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      9542..9677
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(9697..9716)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9698..9716)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9726..9743)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9726..9742)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9735..9757)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9884..10339
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9971..9990)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10181..10202
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        10365..10469
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10470..10538,10539..11330)
                     /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(10688..10707)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11501..12089
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11990..12009
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12243..12260
                     /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 atgacacagt tcgagggctt taccaacctg tatcaggtga gcaagacact gcggtttgag
     3421 ctgatcccac agggcaagac cctgaagcac atccaggagc agggcttcat cgaggaggac
     3481 aaggcccgca atgatcacta caaggagctg aagcccatca tcgatcggat ctacaagacc
     3541 tatgccgacc agtgcctgca gctggtgcag ctggattggg agaacctgag cgccgccatc
     3601 gactcctata gaaaggagaa aaccgaggag acaaggaacg ccctgatcga ggagcaggcc
     3661 acatatcgca atgccatcca cgactacttc atcggccgga cagacaacct gaccgatgcc
     3721 atcaataaga gacacgccga gatctacaag ggcctgttca aggccgagct gtttaatggc
     3781 aaggtgctga agcagctggg caccgtgacc acaaccgagc acgagaacgc cctgctgcgg
     3841 agcttcgaca agtttacaac ctacttctcc ggcttttatg agaacaggaa gaacgtgttc
     3901 agcgccgagg atatcagcac agccatccca caccgcatcg tgcaggacaa cttccccaag
     3961 tttaaggaga attgtcacat cttcacacgc ctgatcaccg ccgtgcccag cctgcgggag
     4021 cactttgaga acgtgaagaa ggccatcggc atcttcgtga gcacctccat cgaggaggtg
     4081 ttttccttcc ctttttataa ccagctgctg acacagaccc agatcgacct gtataaccag
     4141 ctgctgggag gaatctctcg ggaggcaggc accgagaaga tcaagggcct gaacgaggtg
     4201 ctgaatctgg ccatccagaa gaatgatgag acagcccaca tcatcgcctc cctgccacac
     4261 agattcatcc ccctgtttaa gcagatcctg tccgatagga acaccctgtc tttcatcctg
     4321 gaggagttta agagcgacga ggaagtgatc cagtccttct gcaagtacaa gacactgctg
     4381 agaaacgaga acgtgctgga gacagccgag gccctgttta acgagctgaa cagcatcgac
     4441 ctgacacaca tcttcatcag ccacaagaag ctggagacaa tcagcagcgc cctgtgcgac
     4501 cactgggata cactgaggaa tgccctgtat gagcggagaa tctccgagct gacaggcaag
     4561 atcaccaagt ctgccaagga gaaggtgcag cgcagcctga agcacgagga tatcaacctg
     4621 caggagatca tctctgccgc aggcaaggag ctgagcgagg ccttcaagca gaaaaccagc
     4681 gagatcctgt cccacgcaca cgccgccctg gatcagccac tgcctacaac cctgaagaag
     4741 caggaggaga aggagatcct gaagtctcag ctggacagcc tgctgggcct gtaccacctg
     4801 ctggactggt ttgccgtgga tgagtccaac gaggtggacc ccgagttctc tgcccggctg
     4861 accggcatca agctggagat ggagccttct ctgagcttct acaacaaggc cagaaattat
     4921 gccaccaaga agccctactc cgtggagaag ttcaagctga actttcagcg gcctacactg
     4981 gcctctggct gggacgtgaa taaggagaag aacaatggcg ccatcctgtt tgtgaagaac
     5041 ggcctgtact atctgggcat catgccaaag cagaagggca ggtataaggc cctgagcttc
     5101 gagcccacag agaaaaccag cgagggcttt gataagatgt actatgacta cttccctgat
     5161 gccgccaaga tgatcccaaa gtgcagcacc cagctgaagg ccgtgacagc ccactttcag
     5221 acccacacaa cccccatcct gctgtccaac aatttcatcg agcctctgga gatcacaaag
     5281 gagatctacg acctgaacaa tcctgagaag gagccaaaga agtttcagac agcctacgcc
     5341 aagaaaaccg gcgaccagaa gggctacaga gaggccctgt gcaagtggat cgacttcaca
     5401 agggattttc tgtccaagta taccaagaca acctctatcg atctgtctag cctgcggcca
     5461 tcctctcagt ataaggacct gggcgagtac tatgccgagc tgaatcccct gctgtaccac
     5521 atcagcttcc agagaatcgc cgagaaggag atcatggatg ccgtggagac aggcaagctg
     5581 tacctgttcc agatctataa caaggacttt gccaagggcc accacggcaa gcctaatctg
     5641 cacacactgt attggaccgg cctgttttct ccagagaacc tggccaagac aagcatcaag
     5701 ctgaatggcc aggccgagct gttctaccgc cctaagtcca ggatgaagag gatggcacac
     5761 cggctgggag agaagatgct gaacaagaag ctgaaggatc agaaaacccc aatccccgac
     5821 accctgtacc aggagctgta cgactatgtg aatcacagac tgtcccacga cctgtctgat
     5881 gaggccaggg ccctgctgcc caacgtgatc accaaggagg tgtctcacga gatcatcaag
     5941 gataggcgct ttaccagcga caagttcctg ttccacgtgc ctatcacact gaactatcag
     6001 gccgccaatt ccccatctaa gttcaaccag agggtgaatg cctacctgaa ggagcacccc
     6061 gagacaccta tcatcggcat cgatcggggc gagagaaacc tgatctatat cacagtgatc
     6121 gactccaccg gcaagatcct ggagcagcgg agcctgaaca ccatccagca gtttgattac
     6181 cagaagaagc tggacaacag ggagaaggag agggtggcag caaggcaggc ctggtctgtg
     6241 gtgggcacaa tcaaggatct gaagcagggc tatctgagcc aggtcatcca cgagatcgtg
     6301 gacctgatga tccactacca ggccgtggtg gtgctggaga acctgaattt cggctttaag
     6361 agcaagagga ccggcatcgc cgagaaggcc gtgtaccagc agttcgagaa gatgctgatc
     6421 gataagctga attgcctggt gctgaaggac tatccagcag agaaagtggg aggcgtgctg
     6481 aacccatacc agctgacaga ccagttcacc tcctttgcca agatgggcac ccagtctggc
     6541 ttcctgtttt acgtgcctgc cccatataca tctaagatcg atcccctgac cggcttcgtg
     6601 gaccccttcg tgtggaaaac catcaagaat cacgagagcc gcaagcactt cctggagggc
     6661 ttcgactttc tgcactacga cgtgaaaacc ggcgacttca tcctgcactt taagatgaac
     6721 agaaatctgt ccttccagag gggcctgccc ggctttatgc ctgcatggga tatcgtgttc
     6781 gagaagaacg agacacagtt tgacgccaag ggcacccctt tcatcgccgg caagagaatc
     6841 gtgccagtga tcgagaatca cagattcacc ggcagatacc gggacctgta tcctgccaac
     6901 gagctgatcg ccctgctgga ggagaagggc atcgtgttca gggatggctc caacatcctg
     6961 ccaaagctgc tggagaatga cgattctcac gccatcgaca ccatggtggc cctgatccgc
     7021 agcgtgctgc agatgcggaa ctccaatgcc gccacaggcg aggactatat caacagcccc
     7081 gtgcgcgatc tgaatggcgt gtgcttcgac tcccggtttc agaacccaga gtggcccatg
     7141 gacgccgatg ccaatggcgc ctaccacatc gccctgaagg gccagctgct gctgaatcac
     7201 ctgaaggaga gcaaggatct gaagctgcag aacggcatct ccaatcagga ctggctggcc
     7261 tacatccagg agctgcgcaa cggcggctcc cctgctgcta aacgtgttaa gcttgatggg
     7321 ggtagcccgg cagccaagag agtcaaactc gacggcggct ccccagcggc aaaaagggtg
     7381 aaactagacg ggggtagccc cgccgcgaag cgtgtaaagc tggatggagg atcgcctgcg
     7441 gctaagcgag tcaaattaga cggtgggtca ccggctgcaa agcgcgtcaa attggatgct
     7501 agcgaacaaa aactcatctc agaagaggat ctgggcagtg gagagggcag aggaagtctg
     7561 ctaacatgcg gtgacgtcga ggagaatcct ggcccaacca tgattgaaca agatggattg
     7621 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     7681 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     7741 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     7801 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     7861 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     7921 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     7981 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     8041 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     8101 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgatgatct cgtcgtgacc
     8161 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     8221 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     8281 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     8341 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgaacgcgt
     8401 taagtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     8461 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     8521 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     8581 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     8641 acccccactg gttgggacat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     8701 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     8761 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     8821 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     8881 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     8941 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcgt
     9001 cgactttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa
     9061 aggggggact ggaagggcta attcactccc aacgaagaca agatctgctt tttgcttgta
     9121 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     9181 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     9241 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     9301 gcagtacgta tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat
     9361 gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca
     9421 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     9481 ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc
     9541 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     9601 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     9661 aggctttttt ggaggcctag ggacgtaccc aattcgccct atagtgagtc gtattacgcg
     9721 cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     9781 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     9841 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc
     9901 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     9961 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
    10021 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
    10081 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
    10141 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
    10201 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
    10261 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
    10321 atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
    10381 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
    10441 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
    10501 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
    10561 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
    10621 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
    10681 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
    10741 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
    10801 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
    10861 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
    10921 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
    10981 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
    11041 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
    11101 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
    11161 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
    11221 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
    11281 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
    11341 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
    11401 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
    11461 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
    11521 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
    11581 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
    11641 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
    11701 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
    11761 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
    11821 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
    11881 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
    11941 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
    12001 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
    12061 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
    12121 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
    12181 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
    12241 gcagcgagtc agtgagcgag gaagcggaag
//
