LOCUS       sequence-467899-       12181 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of EnAsCas12a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12181)
  AUTHORS   Borck PC, Boyle I, Jankovic K, Bick N, Foster K, Lau AC, 
            Parker-Burns LI, Lubicki DA, Li T, Borah AA, Lofaso NJ, Das Sharma 
            S, Chan T, Kishen RV, Adeagbo A, Raghavan S, Aquilanti E, Prensner 
            JR, Krill-Burger JM, Golub TR, Campbell CD, Dempster JM, Chan EM, 
            Vazquez F
  TITLE     SKI complex loss renders 9p21.3-deleted or MSI-H cancers dependent 
            on PELO.
  JOURNAL   Nature. 2025 Feb 5. doi: 10.1038/s41586-024-08509-3.
  PUBMED    39910293
REFERENCE   2  (bases 1 to 12181)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nature. 
            2025 Feb 5. doi: 10.1038/s41586-024-08509-3."
COMMENT     Sequence Label: pRDA_174_neoR
FEATURES             Location/Qualifiers
     source          1..12181
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      1024..1159
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    1199..1320
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1210..1229)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1264..1283
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     LTR             1392..1625
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    1699..2287
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             1805..1816
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     2214..2234
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2297..3091)
                     /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="MIEQDRLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2408..2427)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3018..3037
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(3098..3151)
                     /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             complement(3167..3247)
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             complement(3254..3301)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(3302..7219)
                     /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
                     NYATKKPYSVEKFKLNFQMPTLARGWDVNREKNNGAILFVKNGLYYLGIMPKQKGRYKA
                     LSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEPL
                     EITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSIDL
                     SSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKGH
                     HGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLKD
                     QKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFFH
                     VPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVIDSTGKILEQRS
                     LNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAVV
                     VLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQF
                     TSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYDV
                     KTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIENH
                     RFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQMR
                     NSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKESK
                     DLKLQNGISNQDWLAYIQELRN"
     regulatory      3573..3582
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(7244..7264)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      7267..7276
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(7285..8462)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          7294..8231
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(7317..7337)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     misc_feature    8614..8729
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     CDS             complement(9007..9051)
                     /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    9236..9469
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    9962..10087
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             10134..10314
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(10315..10541)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     primer_bind     complement(10556..10573)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(10556..10572)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10565..10587)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(10781..10800)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     10900..10922
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(10960..10978)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        11046..11150
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(11151..11219,11220..12011)
                     /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(11369..11388)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga catgattacg aatttggtct ctgcaaaagt
     1021 ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggcc gaggcggcct
     1081 cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg
     1141 gcggagttag gggcgggatg ggcggagtta ggggcgggat agctagagcc agacatgata
     1201 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     1261 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     1321 cctctcactc tctgatattc atttctttgc aagttataaa tactgaataa taagatgaca
     1381 tgaactacta ctgctagaga ttttccacac tgactaaaag ggtctgaggg atctctagtt
     1441 accagagtca cacaacagac gggcacacac tacttgaagc actcaaggca agctttattg
     1501 aggcttaagc agtgggttcc ctagttagcc agagagctcc caggctcaga tctggtctaa
     1561 ccagagagac ccagtacaag caaaaagcag atcttatctt cgttgggagt gaattagccc
     1621 ttccagtccc cccttttctt ttaaaaagtg gctaagatct acagctgcct tgtaagtcat
     1681 tggtcttaaa ggtaccaggc ggggaggcgg cccaaagaga gatccgactc gtctgagggc
     1741 gaaggcggag acgcggaaga ggccgcagag ccggcagcag gccgcgggaa ggaaggtccg
     1801 ctggattgag ggccgaaggg acgtagcaga aggacgtccc gcgcagaatc caggtggcaa
     1861 cacaggcgag cagccaagga aaggacgatg atttccccga caacaccacg gaattgtcag
     1921 tgcccaacag ccgagcccct gtccagcagc gggcaaggca ggcggcgatg agttccgccg
     1981 tggcaatagg gagggggaaa gcgaaagtcc cggaaaggag ctgacaggtg gtggcaatgc
     2041 cccaaccagt gggggttgcg tcagcaaaca cagtgcacac cacgccacgt tgcctgacaa
     2101 cgggccacaa ctcctcataa agagacagca accaggattt atacaaggag gagaaaatga
     2161 aagccatacg ggaagcaata gcatgataca aaggcattaa agcagcgtat ccacatagcg
     2221 taaaaggagc aacatagtta agaataccag tcaatctttc acaaattttg taatccagag
     2281 gttgattgtc gacttatcag aagaactcgt caagaaggcg atagaaggcg atgcgctgcg
     2341 aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg ccgccaagct
     2401 cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc acacccagcc
     2461 ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc ggcaagcagg
     2521 catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gcgcgccttg agcctggcga
     2581 acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga tcgacaagac
     2641 cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc
     2701 aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg gatactttct
     2761 cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc aatagcagcc
     2821 agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg
     2881 ccagccacga tagccgcgct gcctcgtcct gcagttcatt cagggcaccg gacaggtcgg
     2941 tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg gcatcagagc
     3001 agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa gcggccggag
     3061 aacctgcgtg caatctatct tgttcaatca tcccggaagg gccgggattc tcctccacgt
     3121 caccgcatgt tagaaggctt cctctgccct cgcatcctgc ggccgcggca tagtcgggga
     3181 catcatatgg gtatgcataa tcaggcacgt cgtagggata agcgtaatct ggaacatcgt
     3241 atgggtagga tccctttttc ttttttgcct ggccggcctt tttcgtggcc gccggccttt
     3301 tgttgcgcag ctcctggatg taggccagcc agtcctgatt ggagatgccg ttctgcagct
     3361 tcagatcctt gctctccttc aggtgattca gcagcagctg gcccttcagg gcgatgtggt
     3421 aggcgccatt ggcatcggcg tccatgggcc actctgggtt ctgaaaccgg gagtcgaagc
     3481 acacgccatt cagatcgcgc acggggctgt tgatatagtc ctcgcctgtg gcggcattgg
     3541 agttccgcat ctgcagcacg ctgcggatca gggccaccat ggtgtcgatg gcgtgagaat
     3601 cgtcattctc cagcagcttt ggcaggatgt tggagccatc cctgaacacg atgcccttct
     3661 cctccagcag ggcgatcagc tcgttggcag gatacaggtc ccggtatctg ccggtgaatc
     3721 tgtgattctc gatcactggc acgattctct tgccggcgat gaaaggggtg cccttggcgt
     3781 caaactgtgt ctcgttcttc tcgaacacga tatcccatgc aggcataaag ccgggcaggc
     3841 ccctctggaa ggacagattt ctgttcatct taaagtgcag gatgaagtcg ccggttttca
     3901 cgtcgtagtg cagaaagtcg aagccctcca ggaagtgctt gcggctctcg tgattcttga
     3961 tggttttcca cacgaagggg tccacgaagc cggtcagggg atcgatctta gatgtatatg
     4021 gggcaggcac gtaaaacagg aagccagact gggtgcccat cttggcaaag gaggtgaact
     4081 ggtctgtcag ctggtatggg ttcagcacgc ctcccacttt ctctgctgga tagtccttca
     4141 gcaccaggca attcagctta tcgatcagca tcttctcgaa ctgctggtac acggccttct
     4201 cggcgatgcc ggtcctcttg ctcttaaagc cgaaattcag gttctccagc accaccacgg
     4261 cctggtagtg gatcatcagg tccacgatct cgtggatgac ctggctcaga tagccctgct
     4321 tcagatcctt gattgtgccc accacagacc aggcctgcct tgctgccacc ctctccttct
     4381 ccctgttgtc cagcttcttc tggtaatcaa actgctggat ggtgttcagg ctccgctgct
     4441 ccaggatctt gccggtggag tcgatcactg tgatatagat caggtttctc tcgccccgat
     4501 cgatgccgat gataggtgtc tcggggtgct ccttcaggta ggcattcacc ctctggttga
     4561 acttagatgg ggaattggcg gcctgatagt tcagtgtgat aggcacgtgg aaaaagaact
     4621 tgtcgctggt aaagcgccta tccttgatga tctcgtgaga cacctccttg gtgatcacgt
     4681 tgggcagcag ggccctggcc tcatcagaca ggtcgtggga cagtctgtga ttcacatagt
     4741 cgtacagctc ctggtacagg gtgtcgggga ttggggtttt ctgatccttc agcttcttgt
     4801 tcagcatctt ctctcccagc cggtgtgcca tcctcttcat cctggactta gggcggtaga
     4861 acagctcggc ctggccattc agcttgatgc ttgtcttggc caggttctct ggagaaaaca
     4921 ggccggtcca atacagtgtg tgcagattag gcttgccgtg gtggcccttg gcaaagtcct
     4981 tgttatagat ctggaacagg tacagcttgc ctgtctccac ggcatccatg atctccttct
     5041 cggcgattct ctggaagctg atgtggtaca gcaggggatt cagctcggca tagtactcgc
     5101 ccaggtcctt atactgagag gatggccgca ggctagacag atcgatagag gttgtcttgg
     5161 tatacttgga cagaaaatcc cttgtgaagt cgatccactt gcacagggcc tctctgtagc
     5221 ccttctggtc gccggttttc ttggcgtagg ctgtctgaaa cttctttggc tccttctcag
     5281 gattgttcag gtcgtagatc tcctttgtga tctccagagg ctcgatgaaa ttgttggaca
     5341 gcaggatggg ggttgtgtgg gtctgaaagt gggctgtcac ggccttcagc tgggtgctgc
     5401 actttgggat catcttggcg gcatcaggga agtagtcata gtacatctta tcaaagccct
     5461 cgctggtttt ctctgtgggc tcgaagctca gggccttata cctgcccttc tgctttggca
     5521 tgatgcccag atagtacagg ccgttcttca caaacaggat ggcgccattg ttcttctccc
     5581 gattcacgtc ccagccgcgg gccagtgtag gcatctgaaa gttcagcttg aacttctcca
     5641 cggagtaggg cttcttggtg gcataatttc tggccttgtt gtagaagctc agagaaggct
     5701 ccatctccag cttgatgccg gtcagccggg cagagaactc ggggtccacc tcgttggact
     5761 catccacggc aaaccagtcc agcaggtggt acaggcccag caggctgtcc agctgagact
     5821 tcaggatctc cttctcctcc tgcttcttca gggttgtagg cagtggctga tccagggcgg
     5881 cgtgtgcgtg ggacaggatc tcgctggttt tctgcttgaa ggcctcgctc agctccttgc
     5941 ctgcggcaga gatgatctcc tgcaggttga tatcctcgtg cttcaggctg cgctgcacct
     6001 tctccttggc agacttggtg atcttgcctg tcagctcgga gattctccgc tcatacaggg
     6061 cattcctcag tgtatcccag tggtcgcaca gggcgctgct gattgtctcc agcttcttgt
     6121 ggctgatgaa gatgtgtgtc aggtcgatgc tgttcagctc gttaaacagg gcctcggctg
     6181 tctccagcac gttctcgttt ctcagcagtg tcttgtactt gcagaaggac tggatcactt
     6241 cctcgtcgct cttaaactcc tccaggatga aagacagggt gttcctatcg gacaggatct
     6301 gcttaaacag ggggatgaat ctgtgtggca gggaggcgat gatgtgggct gtctcatcat
     6361 tcttctggat ggccagattc agcacctcgt tcaggccctt gatcttctcg gtgcctgcct
     6421 cccgagagat tcctcccagc agctggttat acaggtcgat ctgggtctgt gtcagcagct
     6481 ggttataaaa agggaaggaa aacacctcct cgatggaggt gctcacgaag atgccgatgg
     6541 ccttcttcac gttctcaaag tgctcccgca ggctgggcac ggcggtgatc aggcgtgtga
     6601 agatgtgaca attctcctta aacttgggga agttgtcctg cacgatgcgg tgtgggatgg
     6661 ctgtgctgat atcctcggcg ctgaacacgt tcttcctgtt ccgataaaag ccggagaagt
     6721 aggttgtaaa cttgtcgaag ctccgcagca gggcgttctc gtgctcggtt gtggtcacgg
     6781 tgcccagctg cttcagcacc ttgccattaa acagctcggc cttgaacagg cccttgtaga
     6841 tctcggcgtg tctcttattg atggcatcgg tcaggttgtc tgtccggccg atgaagtagt
     6901 cgtggatggc attgcgatat gtggcctgct cctcgatcag ggcgttcctt gtctcctcgg
     6961 ttttctcctt tctataggag tcgatggcgg cgctcaggtt ctcccaatcc agctgcacca
     7021 gctgcaggca ctggtcggca taggtcttgt agatccgatc gatgatgggc ttcagctcct
     7081 tgtagtgatc attgcgggcc ttgtcctcct cgatgaagcc ctgctcctgg atgtgcttca
     7141 gggtcttgcc ctgtgggatc agctcaaacc gcagtgtctt gctcacctga tacaggttgg
     7201 taaagccctc gaactgtgtg gctgctggga ctccgtggat accgaccttc cgcttcttct
     7261 ttggggccat ggtggcggct agaatcacga cacctgaaat ggaagaaaaa aactttgaac
     7321 cactgtctga ggcttgagaa tgaaccaaga tccaaactca aaaagggcaa attccaagga
     7381 gaattacatc aagtgccaag ctggcctaac ttcagtctcc acccactcag tgtggggaaa
     7441 ctccatcgca taaaacccct ccccccaacc taaagacgac gtactccaaa agctcgagaa
     7501 ctaatcgagg tgcctggacg gcgcccggta ctccgtggag tcacatgaag cgacggctga
     7561 ggacggaaag gcccttttcc tttgtgtggg tgactcaccc gcccgctctc ccgagcgccg
     7621 cgtcctccat tttgagctcc ctgcagcagg gccgggaagc ggccatcttt ccgctcacgc
     7681 aactggtgcc gaccgggcca gccttgccgc ccagggcggg gcgatacacg gcggcgcgag
     7741 gccaggcacc agagcaggcc ggccagcttg agactacccc cgtccgattc tcggtggccg
     7801 cgctcgcagg ccccgcctcg ccgaacatgt gcgctgggac gcacgggccc cgtcgccgcc
     7861 cgcggcccca aaaaccgaaa taccagtgtg cagatcttgg cccgcattta caagactatc
     7921 ttgccagaaa aaaagcgtcg cagcaggtca tcaaaaattt taaatggcta gagacttatc
     7981 gaaagcagcg agacaggcgc gaaggtgcca ccagattcgc acgcggcggc cccagcgccc
     8041 aggccaggcc tcaactcaag cacgaggcga aggggctcct taagcgcaag gcctcgaact
     8101 ctcccaccca cttccaaccc gaagctcggg atcaagaatc acgtactgca gccagtggaa
     8161 gtaattcaag gcacgcaagg gccataaccc gtaaagaggc caggcccgcg ggaaccacac
     8221 acggcactta cctgtgttct ggcggcaaac ccgttgcgaa aaagaacgtt cacggcgact
     8281 actgcactta tatacggttc tcccccaccc tcgggaaaaa ggcggagcca gtacacgaca
     8341 tcactttccc agtttacccc gcgccacctt ctctaggcac cggttcaatt gccgacccct
     8401 ccccccaact tctcggggac tgtgggcgat gtgcgctctg cccactgacg ggcaccggag
     8461 ccaattccca ctcctttcaa gacctagaag gtccattagc tgcaaagatt cctctctgtt
     8521 taaaacttta tccatctttg caactagatt cgaagaaccg gtgatgaatt cagtagcgtg
     8581 aagagcagag aacgagcacc gacaacaacg cagaaatttt gaatttttgt aatttgtttt
     8641 tgtaattctt tagtttgtat gtctgttgct attatgtcta ctattctttc ccctgcactg
     8701 taccccccaa tccccccttt tcttttaaat tgctcgatcc aagcaacgat accgtcgaga
     8761 tccgttcact aatcgaatgg atctgtctct gtctctctct ccaccttctt cttctattcc
     8821 ttcgggcctg tcgggtcccc tcggggttgg gaggtgggtc tgaaacgata atggtgaata
     8881 tccctgccta actctattca ctatagaaag tacagcaaaa actattctta aacctaccaa
     8941 gcctcctact atcattatga ataattttat ataccacagc caatttgtta tgttaaacca
     9001 attccacaaa cttgcccatt tatctaattc caataattct tgttcattct tttcttgctg
     9061 gttttgcgat tcttcaatta aggagtgtat taagcttgtg taattgttaa tttctctgtc
     9121 ccactccatc caggtcgtgt gattccaaat ctgttccaga gatttattac tccaactagc
     9181 attccaaggc acagcagtgg tgcaaatgag ttttccagag caaccccaaa tccccaggag
     9241 ctgttgatcc tttaggtatc tttccacagc caggattctt gcctggagct gcttgatgcc
     9301 ccagactgtg agttgcaaca gatgctgttg cgcctcaata gccctcagca aattgttctg
     9361 ctgctgcact ataccagaca ataattgtct ggcctgtacc gtcagcgtca ttgacgctgc
     9421 gcccatagtg cttcctgctg ctcccaagaa cccaaggaac aaagctccta ttcccactgc
     9481 tcttttttct ctctgcacca ctcttctctt tgccttggtg ggtgctactc ctaatggttc
     9541 aatttttact actttatatt tatataattc acttctccaa ttgtccctca tatctcctcc
     9601 tccaggtctg aagatcagtg gccgcttgct gtgcggtggt cttacttttg ttttgctctt
     9661 cctctatctt gtctaaagct tccttggtgt cttttatctc tatcctttga tgcacacaat
     9721 agagggttgc tactgtatta tataatgatc taagttcttc tgatcctgtc tgaagggatg
     9781 gttgtagctg tcccagtatt tgtctacagc cttctgatgt ttctaacagg ccaggattaa
     9841 ctgcgaatcg ttctagctcc ctgcttgccc atactatatg ttttaattta tattttttct
     9901 ttccccctgg ccttaaccga attttttccc atcgcgatct aattctcccc cgcttaatac
     9961 tgacgctctc gcacccatct ctctccttct agcctccgct agtcaaaatt tttggcgtac
    10021 tcaccagtcg ccgcccctcg cctcttgccg tgcgcgcttc agcaagccga gtcctgcgtc
    10081 gagagagctc ctctggtttc cctttcgctt tcaagtccct gttcgggcgc cactgctaga
    10141 gattttccac actgactaaa agggtctgag ggatctctag ttaccagagt cacacaacag
    10201 acgggcacac actacttgaa gcactcaagg caagctttat tgaggcttaa gcagtgggtt
    10261 ccctagttag ccagagagct cccaggctca gatctggtct aaccagagag acccgtttat
    10321 gtatcgagct aggcacttaa atacaatatc tctgcaatgc ggcaattcag tggttcgtcc
    10381 aatccatgtc agacccgtct gttgccttcc taataaggca cgatcgtacc accttacttc
    10441 caccaatcgg catgcacggt gctttttctc tccttgtaag gcatgttgct aactcatcgt
    10501 taccatgttg caagactaca agagtattgc ataagactac acgcgtcagc ttggcactgg
    10561 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
    10621 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
    10681 cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt ctccttacgc
    10741 atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg
    10801 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
    10861 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
    10921 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
    10981 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
    11041 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
    11101 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
    11161 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
    11221 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
    11281 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
    11341 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
    11401 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
    11461 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
    11521 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
    11581 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
    11641 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
    11701 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
    11761 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
    11821 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
    11881 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
    11941 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
    12001 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
    12061 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
    12121 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
    12181 c
//
