LOCUS       sequence-387210-       11957 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 11957)
  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 11957)
  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: pAR013 Lenti pEF-dEnAsCas12a-NLS-VP64-3XHA-T2A...
FEATURES             Location/Qualifiers
     source          1..11957
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..110
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(148..170)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     270..289
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     483..505
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     497..514
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     498..514
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        529..755
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             756..936
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    983..1108
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1601..1834
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2019..2063
                     /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    2341..2456
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     promoter        2608..3785
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2839..3776
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3733..3753
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3810..3830
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3855..7775
                     /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             7776..7823
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7830..7910
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             7932..8081
                     /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             8091..8144
                     /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             8151..8549
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    8559..9147
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8612..8632)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9030..9041)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9221..9454
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    9526..9647
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9563..9582)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9617..9636
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      9687..9822
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(9855..9871)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9855..9871)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9868..9890)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9879..9895
                     /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(9903..9909,9910..9927,9928..9933))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9948..9969
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10086..10103)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10257..10845)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(11016..11807,11808..11876))
                     /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     11639..11658
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(11877..11957,1..24))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
       61 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
      121 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
      181 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
      241 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
      301 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca
      361 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
      421 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
      481 ttcccagtca cgacgttgta aaacgacggc cagtgccaag ctgacgcgtg tagtcttatg
      541 caatactctt gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc ttacaaggag
      601 agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt gccttattag
      661 gaaggcaaca gacgggtctg acatggattg gacgaaccac tgaattgccg cattgcagag
      721 atattgtatt taagtgccta gctcgataca taaacgggtc tctctggtta gaccagatct
      781 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
      841 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
      901 tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa
      961 agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac
     1021 ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta
     1081 gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg
     1141 ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg
     1201 ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg
     1261 ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag aagaacttag
     1321 atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga
     1381 caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca
     1441 gcaagcggcc actgatcttc agacctggag gaggagatat gagggacaat tggagaagtg
     1501 aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa
     1561 agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt
     1621 tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca
     1681 gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc
     1741 aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg
     1801 ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac
     1861 tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga
     1921 tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa
     1981 tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg
     2041 aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata
     2101 taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac
     2161 tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc
     2221 caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca
     2281 gagacagatc cattcgatta gtgaacggat ctcgacggta tcgttgcttg gatcgagcaa
     2341 tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata
     2401 gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca aaatttctgc
     2461 gttgttgtcg gtgctcgttc tctgctcttc acgctactga attcatcacc ggttcttcga
     2521 atctagttgc aaagatggat aaagttttaa acagagagga atctttgcag ctaatggacc
     2581 ttctaggtct tgaaaggagt gggaattggc tccggtgccc gtcagtgggc agagcgcaca
     2641 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaaccgg tgcctagaga
     2701 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag
     2761 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg
     2821 tttgccgcca gaacacaggt aagtgccgtg tgtggttccc gcgggcctgg cctctttacg
     2881 ggttatggcc cttgcgtgcc ttgaattact tccactggct gcagtacgtg attcttgatc
     2941 ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt
     3001 cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt
     3061 ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat
     3121 gacctgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caagatctgc
     3181 acactggtat ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca
     3241 catgttcggc gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc
     3301 aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg
     3361 cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc
     3421 ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac
     3481 ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt
     3541 accgggcgcc gtccaggcac ctcgattagt tctcgagctt ttggagtacg tcgtctttag
     3601 gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag
     3661 ttaggccagc ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat
     3721 cttggttcat tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt
     3781 cgtgagtgtc gtgagctagc gccaccatgc caaagaagaa gcggaaggtc ggtatccacg
     3841 gagtcccagc agccatgaca cagttcgagg gctttaccaa cctgtatcag gtgagcaaga
     3901 cactgcggtt tgagctgatc ccacagggca agaccctgaa gcacatccag gagcagggct
     3961 tcatcgagga ggacaaggcc cgcaatgatc actacaagga gctgaagccc atcatcgatc
     4021 ggatctacaa gacctatgcc gaccagtgcc tgcagctggt gcagctggat tgggagaacc
     4081 tgagcgccgc catcgactcc tatagaaagg agaaaaccga ggagacaagg aacgccctga
     4141 tcgaggagca ggccacatat cgcaatgcca tccacgacta cttcatcggc cggacagaca
     4201 acctgaccga tgccatcaat aagagacacg ccgagatcta caagggcctg ttcaaggccg
     4261 agctgtttaa tggcaaggtg ctgaagcagc tgggcaccgt gaccacaacc gagcacgaga
     4321 acgccctgct gcggagcttc gacaagttta caacctactt ctccggcttt tatagaaaca
     4381 ggaagaacgt gttcagcgcc gaggatatca gcacagccat cccacaccgc atcgtgcagg
     4441 acaacttccc caagtttaag gagaattgtc acatcttcac acgcctgatc accgccgtgc
     4501 ccagcctgcg ggagcacttt gagaacgtga agaaggccat cggcatcttc gtgagcacct
     4561 ccatcgagga ggtgttttcc ttcccttttt ataaccagct gctgacacag acccagatcg
     4621 acctgtataa ccagctgctg ggaggaatct ctcgggaggc aggcaccgag aagatcaagg
     4681 gcctgaacga ggtgctgaat ctggccatcc agaagaatga tgagacagcc cacatcatcg
     4741 cctccctgcc acacagattc atccccctgt ttaagcagat cctgtccgat aggaacaccc
     4801 tgtctttcat cctggaggag tttaagagcg acgaggaagt gatccagtcc ttctgcaagt
     4861 acaagacact gctgagaaac gagaacgtgc tggagacagc cgaggccctg tttaacgagc
     4921 tgaacagcat cgacctgaca cacatcttca tcagccacaa gaagctggag acaatcagca
     4981 gcgccctgtg cgaccactgg gatacactga ggaatgccct gtatgagcgg agaatctccg
     5041 agctgacagg caagatcacc aagtctgcca aggagaaggt gcagcgcagc ctgaagcacg
     5101 aggatatcaa cctgcaggag atcatctctg ccgcaggcaa ggagctgagc gaggccttca
     5161 agcagaaaac cagcgagatc ctgtcccacg cacacgccgc cctggatcag ccactgccta
     5221 caaccctgaa gaagcaggag gagaaggaga tcctgaagtc tcagctggac agcctgctgg
     5281 gcctgtacca cctgctggac tggtttgccg tggatgagtc caacgaggtg gaccccgagt
     5341 tctctgcccg gctgaccggc atcaagctgg agatggagcc ttctctgagc ttctacaaca
     5401 aggccagaaa ttatgccacc aagaagccct actccgtgga gaagttcaag ctgaactttc
     5461 agatgcctac actggccaga ggctgggacg tgaatagaga gaagaacaat ggcgccatcc
     5521 tgtttgtgaa gaacggcctg tactatctgg gcatcatgcc aaagcagaag ggcaggtata
     5581 aggccctgag cttcgagccc acagagaaaa ccagcgaggg ctttgataag atgtactatg
     5641 actacttccc tgatgccgcc aagatgatcc caaagtgcag cacccagctg aaggccgtga
     5701 cagcccactt tcagacccac acaaccccca tcctgctgtc caacaatttc atcgagcctc
     5761 tggagatcac aaaggagatc tacgacctga acaatcctga gaaggagcca aagaagtttc
     5821 agacagccta cgccaagaaa accggcgacc agaagggcta cagagaggcc ctgtgcaagt
     5881 ggatcgactt cacaagggat tttctgtcca agtataccaa gacaacctct atcgatctgt
     5941 ctagcctgcg gccatcctct cagtataagg acctgggcga gtactatgcc gagctgaatc
     6001 ccctgctgta ccacatcagc ttccagagaa tcgccgagaa ggagatcatg gatgccgtgg
     6061 agacaggcaa gctgtacctg ttccagatct ataacaagga ctttgccaag ggccaccacg
     6121 gcaagcctaa tctgcacaca ctgtattgga ccggcctgtt ttctccagag aacctggcca
     6181 agacaagcat caagctgaat ggccaggccg agctgttcta ccgccctaag tccaggatga
     6241 agaggatggc acaccggctg ggagagaaga tgctgaacaa gaagctgaag gatcagaaaa
     6301 ccccaatccc cgacaccctg taccaggagc tgtacgacta tgtgaatcac agactgtccc
     6361 acgacctgtc tgatgaggcc agggccctgc tgcccaacgt gatcaccaag gaggtgtctc
     6421 acgagatcat caaggatagg cgctttacca gcgacaagtt ctttttccac gtgcctatca
     6481 cactgaacta tcaggccgcc aattccccat ctaagttcaa ccagagggtg aatgcctacc
     6541 tgaaggagca ccccgagaca cctatcatcg gcatcgcccg gggcgagaga aacctgatct
     6601 atatcacagt gatcgactcc accggcaaga tcctggagca gcggagcctg aacaccatcc
     6661 agcagtttga ttaccagaag aagctggaca acagggagaa ggagagggtg gcagcaaggc
     6721 aggcctggtc tgtggtgggc acaatcaagg atctgaagca gggctatctg agccaggtca
     6781 tccacgagat cgtggacctg atgatccact accaggccgt ggtggtgctg gagaacctga
     6841 atttcggctt taagagcaag aggaccggca tcgccgagaa ggccgtgtac cagcagttcg
     6901 agaagatgct gatcgataag ctgaattgcc tggtgctgaa ggactatcca gcagagaaag
     6961 tgggaggcgt gctgaaccca taccagctga cagaccagtt cacctccttt gccaagatgg
     7021 gcacccagtc tggcttcctg ttttacgtgc ctgccccata tacatctaag atcgatcccc
     7081 tgaccggctt cgtggacccc ttcgtgtgga aaaccatcaa gaatcacgag agccgcaagc
     7141 acttcctgga gggcttcgac tttctgcact acgacgtgaa aaccggcgac ttcatcctgc
     7201 actttaagat gaacagaaat ctgtccttcc agaggggcct gcccggcttt atgcctgcat
     7261 gggatatcgt gttcgagaag aacgagacac agtttgacgc caagggcacc cctttcatcg
     7321 ccggcaagag aatcgtgcca gtgatcgaga atcacagatt caccggcaga taccgggacc
     7381 tgtatcctgc caacgagctg atcgccctgc tggaggagaa gggcatcgtg ttcagggatg
     7441 gctccaacat cctgccaaag ctgctggaga atgacgattc tcacgccatc gacacgatgg
     7501 tggccctgat ccgcagcgtg ctgcagatgc ggaactccaa tgccgccaca ggcgaggact
     7561 atatcaacag ccccgtgcgc gatctgaatg gcgtgtgctt cgactcccgg tttcagaacc
     7621 cagagtggcc aatggacgcc gatgccaatg gcgcctacca catcgccctg aagggccagc
     7681 tgctgctgaa tcacctgaag gagagcaagg atctgaagct gcagaacggc atctccaatc
     7741 aggactggct ggcctacatc caggagctgc gcaacaaaag gccggcggcc acgaaaaagg
     7801 ccggccaggc aaaaaagaaa aagggatcct acccatacga tgttccagat tacgcttatc
     7861 cctacgacgt gcctgattat gcatacccat atgatgtccc cgactatgcc ggtggatccg
     7921 gatcaggctc cgacgcattg gacgattttg atctggatat gctgggaagt gacgccctcg
     7981 atgattttga ccttgacatg cttggttcgg atgcccttga tgactttgac ctcgacatgc
     8041 tcggcagtga cgcccttgat gatttcgacc tggacatgct gggatctgga gagggcagag
     8101 gaagccttct aacatgcggt gacgtggagg agaatcccgg cccttccggg atggccaagc
     8161 ctttgtctca agaagaatcc accctcattg aaagagcaac ggctacaatc aacagcatcc
     8221 ccatctctga ggactacagc gtcgccagcg cagctctctc tagcgacggc cgcatcttca
     8281 ctggtgtcaa tgtatatcat tttactgggg gaccttgtgc agaactcgtg gtgctgggca
     8341 ctgctgctgc tgcggcagct ggcaacctga cttgtatcgt cgcgatcgga aatgagaaca
     8401 ggggcatctt gagcccctgc ggacggtgcc gacaggtgct tctcgatctg catcctggga
     8461 tcaaagccat agtgaaggac agtgatggac agccgacggc agttgggatt cgtgaattgc
     8521 tgccctctgg ttatgtgtgg gagggctaat aagtcgacaa tcaacctctg gattacaaaa
     8581 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     8641 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     8701 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     8761 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     8821 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     8881 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     8941 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc
     9001 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     9061 gcggcctgct gccggctctg cggcctcttc cgcgtctccg ccttcgccct cagacgagtc
     9121 ggatctctct ttgggccgcc tccccgcctg gtacctttaa gaccaatgac ttacaaggca
     9181 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc
     9241 caacgaagat aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga
     9301 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
     9361 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
     9421 agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt
     9481 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca
     9541 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     9601 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc
     9661 tagctatccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     9721 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     9781 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt tgcagagacc
     9841 aaattcgtaa tcatgtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     9901 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     9961 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
    10021 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
    10081 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
    10141 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
    10201 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
    10261 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
    10321 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
    10381 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
    10441 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
    10501 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
    10561 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
    10621 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
    10681 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
    10741 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
    10801 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
    10861 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
    10921 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
    10981 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
    11041 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
    11101 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
    11161 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
    11221 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
    11281 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
    11341 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
    11401 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
    11461 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
    11521 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
    11581 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
    11641 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
    11701 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
    11761 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
    11821 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
    11881 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
    11941 tttgaatgta tttagaa
//
