LOCUS       sequence-387119-       11831 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 11831)
  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 11831)
  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: pAR010 Lenti pEF-dEnAsCas12a-NLS-3XHA-T2A-BSD
FEATURES             Location/Qualifiers
     source          1..11831
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        25..251
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             252..432
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    479..604
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1097..1330
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1515..1559
                     /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    1837..1952
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     promoter        2104..3281
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2335..3272
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3229..3249
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3306..3326
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3351..7271
                     /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             7272..7319
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7326..7406
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             7461..7514
                     /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             7521..7919
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    7929..8517
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7982..8002)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8400..8411)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8591..8824
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8896..9017
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8933..8952)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8987..9006
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      9057..9192
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(9225..9241)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9225..9241)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9238..9260)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9249..9265
                     /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(9273..9279,9280..9297,9298..9303))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9318..9339
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9456..9473)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9627..10215)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(10386..11177,11178..11246))
                     /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     11009..11028
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11247..11351)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     11419..11437
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(11475..11497)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     11597..11616
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(11810..11831,1)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(11824..11831,1..10)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     join(11825..11831,1..10)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gacggccagt gccaagctga cgcgtgtagt cttatgcaat actcttgtag tcttgcaaca
       61 tggtaacgat gagttagcaa catgccttac aaggagagaa aaagcaccgt gcatgccgat
      121 tggtggaagt aaggtggtac gatcgtgcct tattaggaag gcaacagacg ggtctgacat
      181 ggattggacg aaccactgaa ttgccgcatt gcagagatat tgtatttaag tgcctagctc
      241 gatacataaa cgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
      301 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
      361 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
      421 aaatctctag cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac cagaggagct
      481 ctctcgacgc aggactcggc ttgctgaagc gcgcacggca agaggcgagg ggcggcgact
      541 ggtgagtacg ccaaaaattt tgactagcgg aggctagaag gagagagatg ggtgcgagag
      601 cgtcagtatt aagcggggga gaattagatc gcgatgggaa aaaattcggt taaggccagg
      661 gggaaagaaa aaatataaat taaaacatat agtatgggca agcagggagc tagaacgatt
      721 cgcagttaat cctggcctgt tagaaacatc agaaggctgt agacaaatac tgggacagct
      781 acaaccatcc cttcagacag gatcagaaga acttagatca ttatataata cagtagcaac
      841 cctctattgt gtgcatcaaa ggatagagat aaaagacacc aaggaagctt tagacaagat
      901 agaggaagag caaaacaaaa gtaagaccac cgcacagcaa gcggccactg atcttcagac
      961 ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa
     1021 aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa
     1081 aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta
     1141 tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc
     1201 agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag
     1261 tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc
     1321 aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt
     1381 ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg
     1441 agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc
     1501 aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt
     1561 ggaattggtt taacataaca aattggctgt ggtatataaa attattcata atgatagtag
     1621 gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc
     1681 agggatattc accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc
     1741 ccgaaggaat agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga
     1801 acggatctcg acggtatcgt tgcttggatc gagcaattta aaagaaaagg ggggattggg
     1861 gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa
     1921 ttacaaaaac aaattacaaa aattcaaaat ttctgcgttg ttgtcggtgc tcgttctctg
     1981 ctcttcacgc tactgaattc atcaccggtt cttcgaatct agttgcaaag atggataaag
     2041 ttttaaacag agaggaatct ttgcagctaa tggaccttct aggtcttgaa aggagtggga
     2101 attggctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg
     2161 gggggagggg tcggcaattg aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa
     2221 agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt
     2281 gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acaggtaagt
     2341 gccgtgtgtg gttcccgcgg gcctggcctc tttacgggtt atggcccttg cgtgccttga
     2401 attacttcca ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt
     2461 gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct
     2521 ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg
     2581 ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct
     2641 ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg
     2701 ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc
     2761 gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc
     2821 ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac
     2881 cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga
     2941 ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc
     3001 cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg
     3061 attagttctc gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga
     3121 tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt
     3181 aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga
     3241 cagtggttca aagttttttt cttccatttc aggtgtcgtg agtgtcgtga gctagcgcca
     3301 ccatgccaaa gaagaagcgg aaggtcggta tccacggagt cccagcagcc atgacacagt
     3361 tcgagggctt taccaacctg tatcaggtga gcaagacact gcggtttgag ctgatcccac
     3421 agggcaagac cctgaagcac atccaggagc agggcttcat cgaggaggac aaggcccgca
     3481 atgatcacta caaggagctg aagcccatca tcgatcggat ctacaagacc tatgccgacc
     3541 agtgcctgca gctggtgcag ctggattggg agaacctgag cgccgccatc gactcctata
     3601 gaaaggagaa aaccgaggag acaaggaacg ccctgatcga ggagcaggcc acatatcgca
     3661 atgccatcca cgactacttc atcggccgga cagacaacct gaccgatgcc atcaataaga
     3721 gacacgccga gatctacaag ggcctgttca aggccgagct gtttaatggc aaggtgctga
     3781 agcagctggg caccgtgacc acaaccgagc acgagaacgc cctgctgcgg agcttcgaca
     3841 agtttacaac ctacttctcc ggcttttata gaaacaggaa gaacgtgttc agcgccgagg
     3901 atatcagcac agccatccca caccgcatcg tgcaggacaa cttccccaag tttaaggaga
     3961 attgtcacat cttcacacgc ctgatcaccg ccgtgcccag cctgcgggag cactttgaga
     4021 acgtgaagaa ggccatcggc atcttcgtga gcacctccat cgaggaggtg ttttccttcc
     4081 ctttttataa ccagctgctg acacagaccc agatcgacct gtataaccag ctgctgggag
     4141 gaatctctcg ggaggcaggc accgagaaga tcaagggcct gaacgaggtg ctgaatctgg
     4201 ccatccagaa gaatgatgag acagcccaca tcatcgcctc cctgccacac agattcatcc
     4261 ccctgtttaa gcagatcctg tccgatagga acaccctgtc tttcatcctg gaggagttta
     4321 agagcgacga ggaagtgatc cagtccttct gcaagtacaa gacactgctg agaaacgaga
     4381 acgtgctgga gacagccgag gccctgttta acgagctgaa cagcatcgac ctgacacaca
     4441 tcttcatcag ccacaagaag ctggagacaa tcagcagcgc cctgtgcgac cactgggata
     4501 cactgaggaa tgccctgtat gagcggagaa tctccgagct gacaggcaag atcaccaagt
     4561 ctgccaagga gaaggtgcag cgcagcctga agcacgagga tatcaacctg caggagatca
     4621 tctctgccgc aggcaaggag ctgagcgagg ccttcaagca gaaaaccagc gagatcctgt
     4681 cccacgcaca cgccgccctg gatcagccac tgcctacaac cctgaagaag caggaggaga
     4741 aggagatcct gaagtctcag ctggacagcc tgctgggcct gtaccacctg ctggactggt
     4801 ttgccgtgga tgagtccaac gaggtggacc ccgagttctc tgcccggctg accggcatca
     4861 agctggagat ggagccttct ctgagcttct acaacaaggc cagaaattat gccaccaaga
     4921 agccctactc cgtggagaag ttcaagctga actttcagat gcctacactg gccagaggct
     4981 gggacgtgaa tagagagaag aacaatggcg ccatcctgtt tgtgaagaac ggcctgtact
     5041 atctgggcat catgccaaag cagaagggca ggtataaggc cctgagcttc gagcccacag
     5101 agaaaaccag cgagggcttt gataagatgt actatgacta cttccctgat gccgccaaga
     5161 tgatcccaaa gtgcagcacc cagctgaagg ccgtgacagc ccactttcag acccacacaa
     5221 cccccatcct gctgtccaac aatttcatcg agcctctgga gatcacaaag gagatctacg
     5281 acctgaacaa tcctgagaag gagccaaaga agtttcagac agcctacgcc aagaaaaccg
     5341 gcgaccagaa gggctacaga gaggccctgt gcaagtggat cgacttcaca agggattttc
     5401 tgtccaagta taccaagaca acctctatcg atctgtctag cctgcggcca tcctctcagt
     5461 ataaggacct gggcgagtac tatgccgagc tgaatcccct gctgtaccac atcagcttcc
     5521 agagaatcgc cgagaaggag atcatggatg ccgtggagac aggcaagctg tacctgttcc
     5581 agatctataa caaggacttt gccaagggcc accacggcaa gcctaatctg cacacactgt
     5641 attggaccgg cctgttttct ccagagaacc tggccaagac aagcatcaag ctgaatggcc
     5701 aggccgagct gttctaccgc cctaagtcca ggatgaagag gatggcacac cggctgggag
     5761 agaagatgct gaacaagaag ctgaaggatc agaaaacccc aatccccgac accctgtacc
     5821 aggagctgta cgactatgtg aatcacagac tgtcccacga cctgtctgat gaggccaggg
     5881 ccctgctgcc caacgtgatc accaaggagg tgtctcacga gatcatcaag gataggcgct
     5941 ttaccagcga caagttcttt ttccacgtgc ctatcacact gaactatcag gccgccaatt
     6001 ccccatctaa gttcaaccag agggtgaatg cctacctgaa ggagcacccc gagacaccta
     6061 tcatcggcat cgcccggggc gagagaaacc tgatctatat cacagtgatc gactccaccg
     6121 gcaagatcct ggagcagcgg agcctgaaca ccatccagca gtttgattac cagaagaagc
     6181 tggacaacag ggagaaggag agggtggcag caaggcaggc ctggtctgtg gtgggcacaa
     6241 tcaaggatct gaagcagggc tatctgagcc aggtcatcca cgagatcgtg gacctgatga
     6301 tccactacca ggccgtggtg gtgctggaga acctgaattt cggctttaag agcaagagga
     6361 ccggcatcgc cgagaaggcc gtgtaccagc agttcgagaa gatgctgatc gataagctga
     6421 attgcctggt gctgaaggac tatccagcag agaaagtggg aggcgtgctg aacccatacc
     6481 agctgacaga ccagttcacc tcctttgcca agatgggcac ccagtctggc ttcctgtttt
     6541 acgtgcctgc cccatataca tctaagatcg atcccctgac cggcttcgtg gaccccttcg
     6601 tgtggaaaac catcaagaat cacgagagcc gcaagcactt cctggagggc ttcgactttc
     6661 tgcactacga cgtgaaaacc ggcgacttca tcctgcactt taagatgaac agaaatctgt
     6721 ccttccagag gggcctgccc ggctttatgc ctgcatggga tatcgtgttc gagaagaacg
     6781 agacacagtt tgacgccaag ggcacccctt tcatcgccgg caagagaatc gtgccagtga
     6841 tcgagaatca cagattcacc ggcagatacc gggacctgta tcctgccaac gagctgatcg
     6901 ccctgctgga ggagaagggc atcgtgttca gggatggctc caacatcctg ccaaagctgc
     6961 tggagaatga cgattctcac gccatcgaca cgatggtggc cctgatccgc agcgtgctgc
     7021 agatgcggaa ctccaatgcc gccacaggcg aggactatat caacagcccc gtgcgcgatc
     7081 tgaatggcgt gtgcttcgac tcccggtttc agaacccaga gtggccaatg gacgccgatg
     7141 ccaatggcgc ctaccacatc gccctgaagg gccagctgct gctgaatcac ctgaaggaga
     7201 gcaaggatct gaagctgcag aacggcatct ccaatcagga ctggctggcc tacatccagg
     7261 agctgcgcaa caaaaggccg gcggccacga aaaaggccgg ccaggcaaaa aagaaaaagg
     7321 gatcctaccc atacgatgtt ccagattacg cttatcccta cgacgtgcct gattatgcat
     7381 acccatatga tgtccccgac tatgccggtg gatccggatc aggctccggg tcttcggttc
     7441 aaggaagacg tggatctgga gagggcagag gaagccttct aacatgcggt gacgtggagg
     7501 agaatcccgg cccttccggg atggccaagc ctttgtctca agaagaatcc accctcattg
     7561 aaagagcaac ggctacaatc aacagcatcc ccatctctga ggactacagc gtcgccagcg
     7621 cagctctctc tagcgacggc cgcatcttca ctggtgtcaa tgtatatcat tttactgggg
     7681 gaccttgtgc agaactcgtg gtgctgggca ctgctgctgc tgcggcagct ggcaacctga
     7741 cttgtatcgt cgcgatcgga aatgagaaca ggggcatctt gagcccctgc ggacggtgcc
     7801 gacaggtgct tctcgatctg catcctggga tcaaagccat agtgaaggac agtgatggac
     7861 agccgacggc agttgggatt cgtgaattgc tgccctctgg ttatgtgtgg gagggctaat
     7921 aagtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     7981 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     8041 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     8101 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     8161 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     8221 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     8281 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
     8341 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     8401 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     8461 cgcgtctccg ccttcgccct cagacgagtc ggatctctct ttgggccgcc tccccgcctg
     8521 gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa
     8581 aaggggggac tggaagggct aattcactcc caacgaagat aagatctgct ttttgcttgt
     8641 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     8701 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     8761 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     8821 agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata
     8881 tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     8941 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     9001 ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccatcc
     9061 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     9121 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     9181 tttttggagg cctagacttt tgcagagacc aaattcgtaa tcatgtcata gctgtttcct
     9241 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     9301 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     9361 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     9421 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     9481 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     9541 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     9601 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     9661 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     9721 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     9781 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     9841 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     9901 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     9961 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
    10021 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
    10081 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
    10141 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
    10201 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
    10261 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
    10321 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
    10381 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
    10441 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
    10501 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
    10561 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
    10621 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
    10681 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
    10741 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
    10801 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
    10861 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
    10921 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
    10981 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
    11041 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
    11101 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
    11161 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
    11221 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
    11281 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
    11341 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
    11401 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
    11461 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
    11521 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
    11581 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt
    11641 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc gccattcagg
    11701 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
    11761 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
    11821 cgttgtaaaa c
//
