LOCUS       sequence-387207-       13376 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dCas12a effector fusion for manipulating transcription.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13376)
  AUTHORS   Tycko J, Van MV, Aradhana, DelRosso N, Ye H, Yao D, Valbuena R, 
            Vaughan-Jackson A, Xu X, Ludwig C, Spees K, Liu K, Gu M, Khare V, 
            Mukund AX, Suzuki PH, Arana S, Zhang C, Du PP, Ornstein TS, Hess GT,
            Kamber RA, Qi LS, Khalil AS, Bintu L, Bassik MC
  TITLE     Development of compact transcriptional effectors using 
            high-throughput measurements in diverse contexts.
  JOURNAL   Nat Biotechnol. 2024 Nov 1. doi: 10.1038/s41587-024-02442-6.
  PUBMED    39487265
REFERENCE   2  (bases 1 to 13376)
  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: "Nat 
            Biotechnol. 2024 Nov 1. doi: 10.1038/s41587-024-02442-6."
COMMENT     Sequence Label: pAR014 Lenti pEF-dEnAsCas12a-NLS-VPR-3XHA-T2A-BSD
FEATURES             Location/Qualifiers
     source          1..13376
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(321..1112,1113..1181))
                     /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     944..963
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1182..1286)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1354..1372
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1410..1432)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1532..1551
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1745..1767
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1759..1776
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1760..1776
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1791..2017
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             2018..2198
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2245..2370
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2863..3096
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3281..3325
                     /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    3603..3718
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     promoter        3870..5047
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          4101..5038
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     4995..5015
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             5072..5092
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5117..9037
                     /codon_start=1
                     /product="CRISPR-associated protein from Acidaminococcus 
                     sp. BV3L6 (Zetsche et al., 2015)"
                     /label=AsCpf1
                     /translation="MTQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARND
                     HYKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRN
                     AIHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFD
                     KFTTYFSGFYRNRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHF
                     ENVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVL
                     NLAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLL
                     RNENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTG
                     KITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTL
                     KKQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKA
                     RNYATKKPYSVEKFKLNFQMPTLARGWDVNREKNNGAILFVKNGLYYLGIMPKQKGRYK
                     ALSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEP
                     LEITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSID
                     LSSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKG
                     HHGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLK
                     DQKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFF
                     HVPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIARGERNLIYITVIDSTGKILEQR
                     SLNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAV
                     VVLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQ
                     FTSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYD
                     VKTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIEN
                     HRFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQM
                     RNSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKES
                     KDLKLQNGISNQDWLAYIQELRN"
     CDS             9038..9085
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             9092..9172
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             9194..9343
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             9368..9388
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9818..10174
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALL"
     CDS             10193..10762
                     /codon_start=1
                     /product="transcriptional activation domain from the human 
                     herpesvirus 4 (Epstein-Barr virus) replication and 
                     transcription activator Rta/BRLF1 (Hardwick et al., 1992; 
                     Chavez et al., 2015)"
                     /label=Rta AD
                     /translation="RDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWAN
                     RPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKAL
                     REMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEIL
                     DTFLNDECLLHAMHISTGLSIFDTSLF"
     CDS             10772..10825
                     /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             10832..11230
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    11240..11828
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(11293..11313)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(11711..11722)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             11902..12135
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    12207..12328
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12244..12263)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12298..12317
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      12368..12503
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(12536..12552)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12536..12552)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12549..12571)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12560..12576
                     /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)."
     promoter        complement(join(12584..12590,12591..12608,12609..12614))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12629..12650
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12767..12784)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(12938..13376,1..150))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
       61 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
      121 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
      181 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
      241 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
      301 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
      361 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
      421 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
      481 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
      541 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
      601 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
      661 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
      721 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
      781 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
      841 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
      901 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
      961 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     1021 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     1081 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     1141 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     1201 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     1261 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     1321 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     1381 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     1441 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     1501 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     1561 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
     1621 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     1681 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     1741 ttttcccagt cacgacgttg taaaacgacg gccagtgcca agctgacgcg tgtagtctta
     1801 tgcaatactc ttgtagtctt gcaacatggt aacgatgagt tagcaacatg ccttacaagg
     1861 agagaaaaag caccgtgcat gccgattggt ggaagtaagg tggtacgatc gtgccttatt
     1921 aggaaggcaa cagacgggtc tgacatggat tggacgaacc actgaattgc cgcattgcag
     1981 agatattgta tttaagtgcc tagctcgata cataaacggg tctctctggt tagaccagat
     2041 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
     2101 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
     2161 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg
     2221 aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc
     2281 acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc
     2341 tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga
     2401 tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta
     2461 tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa
     2521 ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt
     2581 agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa
     2641 gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca
     2701 cagcaagcgg ccactgatct tcagacctgg aggaggagat atgagggaca attggagaag
     2761 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     2821 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     2881 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     2941 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     3001 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     3061 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     3121 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     3181 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     3241 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     3301 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     3361 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     3421 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     3481 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     3541 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgttgct tggatcgagc
     3601 aatttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa
     3661 tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaatttct
     3721 gcgttgttgt cggtgctcgt tctctgctct tcacgctact gaattcatca ccggttcttc
     3781 gaatctagtt gcaaagatgg ataaagtttt aaacagagag gaatctttgc agctaatgga
     3841 ccttctaggt cttgaaagga gtgggaattg gctccggtgc ccgtcagtgg gcagagcgca
     3901 catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacc ggtgcctaga
     3961 gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg
     4021 agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg
     4081 ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc ccgcgggcct ggcctcttta
     4141 cgggttatgg cccttgcgtg ccttgaatta cttccactgg ctgcagtacg tgattcttga
     4201 tcccgagctt cgggttggaa gtgggtggga gagttcgagg ccttgcgctt aaggagcccc
     4261 ttcgcctcgt gcttgagttg aggcctggcc tgggcgctgg ggccgccgcg tgcgaatctg
     4321 gtggcacctt cgcgcctgtc tcgctgcttt cgataagtct ctagccattt aaaatttttg
     4381 atgacctgct gcgacgcttt ttttctggca agatagtctt gtaaatgcgg gccaagatct
     4441 gcacactggt atttcggttt ttggggccgc gggcggcgac ggggcccgtg cgtcccagcg
     4501 cacatgttcg gcgaggcggg gcctgcgagc gcggccaccg agaatcggac gggggtagtc
     4561 tcaagctggc cggcctgctc tggtgcctgg cctcgcgccg ccgtgtatcg ccccgccctg
     4621 ggcggcaagg ctggcccggt cggcaccagt tgcgtgagcg gaaagatggc cgcttcccgg
     4681 ccctgctgca gggagctcaa aatggaggac gcggcgctcg ggagagcggg cgggtgagtc
     4741 acccacacaa aggaaaaggg cctttccgtc ctcagccgtc gcttcatgtg actccacgga
     4801 gtaccgggcg ccgtccaggc acctcgatta gttctcgagc ttttggagta cgtcgtcttt
     4861 aggttggggg gaggggtttt atgcgatgga gtttccccac actgagtggg tggagactga
     4921 agttaggcca gcttggcact tgatgtaatt ctccttggaa tttgcccttt ttgagtttgg
     4981 atcttggttc attctcaagc ctcagacagt ggttcaaagt ttttttcttc catttcaggt
     5041 gtcgtgagtg tcgtgagcta gcgccaccat gccaaagaag aagcggaagg tcggtatcca
     5101 cggagtccca gcagccatga cacagttcga gggctttacc aacctgtatc aggtgagcaa
     5161 gacactgcgg tttgagctga tcccacaggg caagaccctg aagcacatcc aggagcaggg
     5221 cttcatcgag gaggacaagg cccgcaatga tcactacaag gagctgaagc ccatcatcga
     5281 tcggatctac aagacctatg ccgaccagtg cctgcagctg gtgcagctgg attgggagaa
     5341 cctgagcgcc gccatcgact cctatagaaa ggagaaaacc gaggagacaa ggaacgccct
     5401 gatcgaggag caggccacat atcgcaatgc catccacgac tacttcatcg gccggacaga
     5461 caacctgacc gatgccatca ataagagaca cgccgagatc tacaagggcc tgttcaaggc
     5521 cgagctgttt aatggcaagg tgctgaagca gctgggcacc gtgaccacaa ccgagcacga
     5581 gaacgccctg ctgcggagct tcgacaagtt tacaacctac ttctccggct tttatagaaa
     5641 caggaagaac gtgttcagcg ccgaggatat cagcacagcc atcccacacc gcatcgtgca
     5701 ggacaacttc cccaagttta aggagaattg tcacatcttc acacgcctga tcaccgccgt
     5761 gcccagcctg cgggagcact ttgagaacgt gaagaaggcc atcggcatct tcgtgagcac
     5821 ctccatcgag gaggtgtttt ccttcccttt ttataaccag ctgctgacac agacccagat
     5881 cgacctgtat aaccagctgc tgggaggaat ctctcgggag gcaggcaccg agaagatcaa
     5941 gggcctgaac gaggtgctga atctggccat ccagaagaat gatgagacag cccacatcat
     6001 cgcctccctg ccacacagat tcatccccct gtttaagcag atcctgtccg ataggaacac
     6061 cctgtctttc atcctggagg agtttaagag cgacgaggaa gtgatccagt ccttctgcaa
     6121 gtacaagaca ctgctgagaa acgagaacgt gctggagaca gccgaggccc tgtttaacga
     6181 gctgaacagc atcgacctga cacacatctt catcagccac aagaagctgg agacaatcag
     6241 cagcgccctg tgcgaccact gggatacact gaggaatgcc ctgtatgagc ggagaatctc
     6301 cgagctgaca ggcaagatca ccaagtctgc caaggagaag gtgcagcgca gcctgaagca
     6361 cgaggatatc aacctgcagg agatcatctc tgccgcaggc aaggagctga gcgaggcctt
     6421 caagcagaaa accagcgaga tcctgtccca cgcacacgcc gccctggatc agccactgcc
     6481 tacaaccctg aagaagcagg aggagaagga gatcctgaag tctcagctgg acagcctgct
     6541 gggcctgtac cacctgctgg actggtttgc cgtggatgag tccaacgagg tggaccccga
     6601 gttctctgcc cggctgaccg gcatcaagct ggagatggag ccttctctga gcttctacaa
     6661 caaggccaga aattatgcca ccaagaagcc ctactccgtg gagaagttca agctgaactt
     6721 tcagatgcct acactggcca gaggctggga cgtgaataga gagaagaaca atggcgccat
     6781 cctgtttgtg aagaacggcc tgtactatct gggcatcatg ccaaagcaga agggcaggta
     6841 taaggccctg agcttcgagc ccacagagaa aaccagcgag ggctttgata agatgtacta
     6901 tgactacttc cctgatgccg ccaagatgat cccaaagtgc agcacccagc tgaaggccgt
     6961 gacagcccac tttcagaccc acacaacccc catcctgctg tccaacaatt tcatcgagcc
     7021 tctggagatc acaaaggaga tctacgacct gaacaatcct gagaaggagc caaagaagtt
     7081 tcagacagcc tacgccaaga aaaccggcga ccagaagggc tacagagagg ccctgtgcaa
     7141 gtggatcgac ttcacaaggg attttctgtc caagtatacc aagacaacct ctatcgatct
     7201 gtctagcctg cggccatcct ctcagtataa ggacctgggc gagtactatg ccgagctgaa
     7261 tcccctgctg taccacatca gcttccagag aatcgccgag aaggagatca tggatgccgt
     7321 ggagacaggc aagctgtacc tgttccagat ctataacaag gactttgcca agggccacca
     7381 cggcaagcct aatctgcaca cactgtattg gaccggcctg ttttctccag agaacctggc
     7441 caagacaagc atcaagctga atggccaggc cgagctgttc taccgcccta agtccaggat
     7501 gaagaggatg gcacaccggc tgggagagaa gatgctgaac aagaagctga aggatcagaa
     7561 aaccccaatc cccgacaccc tgtaccagga gctgtacgac tatgtgaatc acagactgtc
     7621 ccacgacctg tctgatgagg ccagggccct gctgcccaac gtgatcacca aggaggtgtc
     7681 tcacgagatc atcaaggata ggcgctttac cagcgacaag ttctttttcc acgtgcctat
     7741 cacactgaac tatcaggccg ccaattcccc atctaagttc aaccagaggg tgaatgccta
     7801 cctgaaggag caccccgaga cacctatcat cggcatcgcc cggggcgaga gaaacctgat
     7861 ctatatcaca gtgatcgact ccaccggcaa gatcctggag cagcggagcc tgaacaccat
     7921 ccagcagttt gattaccaga agaagctgga caacagggag aaggagaggg tggcagcaag
     7981 gcaggcctgg tctgtggtgg gcacaatcaa ggatctgaag cagggctatc tgagccaggt
     8041 catccacgag atcgtggacc tgatgatcca ctaccaggcc gtggtggtgc tggagaacct
     8101 gaatttcggc tttaagagca agaggaccgg catcgccgag aaggccgtgt accagcagtt
     8161 cgagaagatg ctgatcgata agctgaattg cctggtgctg aaggactatc cagcagagaa
     8221 agtgggaggc gtgctgaacc cataccagct gacagaccag ttcacctcct ttgccaagat
     8281 gggcacccag tctggcttcc tgttttacgt gcctgcccca tatacatcta agatcgatcc
     8341 cctgaccggc ttcgtggacc ccttcgtgtg gaaaaccatc aagaatcacg agagccgcaa
     8401 gcacttcctg gagggcttcg actttctgca ctacgacgtg aaaaccggcg acttcatcct
     8461 gcactttaag atgaacagaa atctgtcctt ccagaggggc ctgcccggct ttatgcctgc
     8521 atgggatatc gtgttcgaga agaacgagac acagtttgac gccaagggca cccctttcat
     8581 cgccggcaag agaatcgtgc cagtgatcga gaatcacaga ttcaccggca gataccggga
     8641 cctgtatcct gccaacgagc tgatcgccct gctggaggag aagggcatcg tgttcaggga
     8701 tggctccaac atcctgccaa agctgctgga gaatgacgat tctcacgcca tcgacacgat
     8761 ggtggccctg atccgcagcg tgctgcagat gcggaactcc aatgccgcca caggcgagga
     8821 ctatatcaac agccccgtgc gcgatctgaa tggcgtgtgc ttcgactccc ggtttcagaa
     8881 cccagagtgg ccaatggacg ccgatgccaa tggcgcctac cacatcgccc tgaagggcca
     8941 gctgctgctg aatcacctga aggagagcaa ggatctgaag ctgcagaacg gcatctccaa
     9001 tcaggactgg ctggcctaca tccaggagct gcgcaacaaa aggccggcgg ccacgaaaaa
     9061 ggccggccag gcaaaaaaga aaaagggatc ctacccatac gatgttccag attacgctta
     9121 tccctacgac gtgcctgatt atgcataccc atatgatgtc cccgactatg ccggtggatc
     9181 cggatcaggc tccgacgcat tggacgattt tgatctggat atgctgggaa gtgacgccct
     9241 cgatgatttt gaccttgaca tgcttggttc ggatgccctt gatgactttg acctcgacat
     9301 gctcggcagt gacgcccttg atgatttcga cctggacatg ctgattaact ctagaagttc
     9361 cggatctccg aaaaagaaac gcaaagttgg tagccagtac ctgcccgaca ccgacgaccg
     9421 gcaccggatc gaggaaaagc ggaagcggac ctacgagaca ttcaagagca tcatgaagaa
     9481 gtcccccttc agcggcccca ccgaccctag acctccacct agaagaatcg ccgtgcccag
     9541 cagatccagc gccagcgtgc caaaacctgc cccccagcct taccccttca ccagcagcct
     9601 gagcaccatc aactacgacg agttccctac catggtgttc cccagcggcc agatctctca
     9661 ggcctctgct ctggctccag cccctcctca ggtgctgcct caggctcctg ctcctgcacc
     9721 agctccagcc atggtgtctg cactggctca ggcaccagca cccgtgcctg tgctggctcc
     9781 tggacctcca caggctgtgg ctccaccagc ccctaaacct acacaggccg gcgagggcac
     9841 actgtctgaa gctctgctgc agctgcagtt cgacgacgag gatctgggag ccctgctggg
     9901 aaacagcacc gatcctgccg tgttcaccga cctggccagc gtggacaaca gcgagttcca
     9961 gcagctgctg aaccagggca tccctgtggc ccctcacacc accgagccca tgctgatgga
    10021 ataccccgag gccatcaccc ggctcgtgac aggcgctcag aggcctcctg atccagctcc
    10081 tgcccctctg ggagcaccag gcctgcctaa tggactgctg tctggcgacg aggacttcag
    10141 ctctatcgcc gatatggatt tctcagcctt gctgggctct ggcagcggca gccgggattc
    10201 cagggaaggg atgtttttgc cgaagcctga ggccggctcc gctattagtg acgtgtttga
    10261 gggccgcgag gtgtgccagc caaaacgaat ccggccattt catcctccag gaagtccatg
    10321 ggccaaccgc ccactccccg ccagcctcgc accaacacca accggtccag tacatgagcc
    10381 agtcgggtca ctgaccccgg caccagtccc tcagccactg gatccagcgc ccgcagtgac
    10441 tcccgaggcc agtcacctgt tggaggatcc cgatgaagag acgagccagg ctgtcaaagc
    10501 ccttcgggag atggccgata ctgtgattcc ccagaaggaa gaggctgcaa tctgtggcca
    10561 aatggacctt tcccatccgc ccccaagggg ccatctggat gagctgacaa ccacacttga
    10621 gtccatgacc gaggatctga acctggactc acccctgacc ccggaattga acgagattct
    10681 ggataccttc ctgaacgacg agtgcctctt gcatgccatg catatcagca caggactgtc
    10741 catcttcgac acatctctgt ttggatctgg agagggcaga ggaagccttc taacatgcgg
    10801 tgacgtggag gagaatcccg gcccttccgg gatggccaag cctttgtctc aagaagaatc
    10861 caccctcatt gaaagagcaa cggctacaat caacagcatc cccatctctg aggactacag
    10921 cgtcgccagc gcagctctct ctagcgacgg ccgcatcttc actggtgtca atgtatatca
    10981 ttttactggg ggaccttgtg cagaactcgt ggtgctgggc actgctgctg ctgcggcagc
    11041 tggcaacctg acttgtatcg tcgcgatcgg aaatgagaac aggggcatct tgagcccctg
    11101 cggacggtgc cgacaggtgc ttctcgatct gcatcctggg atcaaagcca tagtgaagga
    11161 cagtgatgga cagccgacgg cagttgggat tcgtgaattg ctgccctctg gttatgtgtg
    11221 ggagggctaa taagtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg
    11281 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
    11341 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
    11401 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
    11461 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
    11521 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
    11581 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
    11641 gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
    11701 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
    11761 gcggcctctt ccgcgtctcc gccttcgccc tcagacgagt cggatctctc tttgggccgc
    11821 ctccccgcct ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact
    11881 ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga taagatctgc
    11941 tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct
    12001 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
    12061 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
    12121 ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa
    12181 agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat
    12241 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
    12301 gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac
    12361 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
    12421 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
    12481 gtgaggaggc ttttttggag gcctagactt ttgcagagac caaattcgta atcatgtcat
    12541 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
    12601 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
    12661 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
    12721 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
    12781 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
    12841 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
    12901 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
    12961 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
    13021 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
    13081 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
    13141 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
    13201 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
    13261 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
    13321 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actaga
//
