LOCUS       sequence-496435-       12953 bp    DNA     circular SYN 07-JAN-2026
DEFINITION  Overexpression of EnAsCas12a-BFP with HBB IVS2 intron in 5' UTR with
            blasticidin resistance gene. Contains minimal A2UCOE and Cp36 
            recombination site..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12953)
  AUTHORS   Arana S, Du PP, Vaughan-Jackson A, Enright N, Spees K, Valbuena R, 
            Garcia CA, Nguyen T, Venida A, Seczynska M, Bintu L, Lehner PJ, 
            Prolo LM, Bassik MC
  TITLE     Reduced Cas9 transgene silencing by incorporation of intron 
            sequences.
  JOURNAL   Nat Commun. 2025 Nov 27;16(1):10656. doi: 
            10.1038/s41467-025-65669-0.
  PUBMED    41309578
REFERENCE   2  (bases 1 to 12953)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 7, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2025-11-27"; volume: "16(1):10656. doi"; pages: " 
            10.1038/s41467-025-65669-0"
COMMENT     Sequence Label: pPD687 A2UCOE-HBB_IVS2-EnAsCas12a-BFP
FEATURES             Location/Qualifiers
     source          1..12953
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        500..1709
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     1546..1565
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             1713..2111
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    2138..2186
                     /note="synthetic polyadenylation signal"
     CDS             join(2452..2520,2521..3312)
                     /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(2670..2689)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3325..3461
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3613..3684
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3689..3735)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3753..3801
                     /note="synthetic polyadenylation signal"
     promoter        4627..5805
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          4858..5796
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     5753..5773
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      6787..6796
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6799..6819
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6844..10761
                     /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      10481..10490
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             10762..10809
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             10825..11523
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     polyA_signal    11575..11696
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11586..11605)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     11640..11659
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      11811..11857
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      11887..12023
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    12068..12139
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(12365..12953)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12445..12464)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
       61 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
      181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcattt
      241 aaatttccga actctccaag gccctcgtcg gaaaatcttc aaacctttcg tccgatccat
      301 cttgcaggct acctctcgaa cgaactatcg caagtctctt ggccggcctt gcgccttggc
      361 tattgcttgg cagcgcctat cgccaggtat tactccaatc ccgaatatcc gagatcggga
      421 tcacccgaga gaagttcaac ctacatcctc aatcccgatc tatccgagat ccgaggaata
      481 tcgaaatcgg ggcgcgcctg gcctccgcgc cgggttttgg cgcctcccgc gggcgccccc
      541 ctcgtcacgg cgagcgctgc cacgtcagac gaagggcgca ggagcgtcct gatccttccg
      601 cccggacgct caggacagcg gcccgctgct cataagactc ggccttagaa ccccagtatc
      661 agcagaagga cattttagga cgggacttgg gtgactctag ggcactggtt ttctttccag
      721 agagcggaac aggcgaggaa aagtagtccc ttctcggcga ttctgcggag ggatctccgt
      781 ggggcggtga acgccgatga ttatataagg acgcgccggg tgtggcacag ctagttccgt
      841 cgcagccggg atttgggtcg cggttcttgt ttgtggatcg ctgtgatcgt cacttggtga
      901 gtagcgggct gctgggctgg ccggggcttt cgtggccgcc gggccgctcg gtgggacgga
      961 agcgtgtgga gagaccgcca agggctgtag tctgggtccg cgagcaaggt tgccctgaac
     1021 tgggggttgg ggggagcgca gcaaaatggc ggctgttccc gagtcttgaa tggaagacgc
     1081 ttgtgaggcg ggctgtgagg tcgttgaaac aaggtggggg gcatggtggg cggcaagaac
     1141 ccaaggtctt gagcccttcg ctaatgcggg aaagctctta ttcgggtgag atgggctggg
     1201 caccatctgg ggaccctgac gtgaagtttg tcactgactg gagaactcgg tttgtcgtct
     1261 gttgcggggg cggcagttat ggcggtgccg ttgggcagtg cacccgtacc tttgggagcg
     1321 cgcgccctcg tcgtgtcgtg acgtcacccg ttctgttggc ttataatgca gggtggggcc
     1381 acctgccggt aggtgtgcgg taggcttttc tccgtcgcag gacgcagggt tcgggcctag
     1441 ggtaggctct cctgaatcga caggcgccgg acctctggtg aggggaggga taagtgaggc
     1501 gtcagtttct ttggtcggtt ttatgtacct atcttcttaa gtagctgaag ctccggtttt
     1561 gaactatgcg ctcggggttg gcgagtgtgt tttgtgaagt tttttaggca ccttttgaaa
     1621 tgtaatcatt tgggtcaata tgtaattttc agtgttagac ttgtaaattg tccgctaaat
     1681 tctggccgtt tttggctttt ttgttagaca acatggccaa gcctttgtct caagaagaat
     1741 ccaccctcat tgaaagagca acggctacaa tcaacagcat ccccatctct gaagactaca
     1801 gcgtcgccag cgcagctctc tctagcgacg gccgcatctt cactggtgtc aatgtatatc
     1861 attttactgg gggaccttgt gcagaactcg tggtgctggg cactgctgct gctgcggcag
     1921 ctggcaacct gacttgtatc gtcgcgatcg gaaatgagaa caggggcatc ttgagcccct
     1981 gcggacggtg ccgacaggtg cttctcgatc tgcatcctgg gatcaaagcc atagtgaagg
     2041 acagtgatgg acagccgacg gcagttggga ttcgtgaatt gctgccctct ggttatgtgt
     2101 gggagggcta agctagtatg taagcctagt cttagataat aaaatcgcta tccatcgaag
     2161 atggatgtgt gttggttttt tgtgtgtgta acgctaggcg cgcctggtgt accgagaacg
     2221 atcctctcag tgcgagtctc gacgatccat atcgttgctt ggcagtcagc cagtcggaat
     2281 ccagcttggg acccaggaag tccaatcgtc agatattgta ctcaagcctg gtcacggcag
     2341 cgtaccgatc tgtttaaacc tagatattga tagtctgatc ggtcaacgta taatcgagtc
     2401 ctagcttttg caaacatcta tcaagagaca ggatcagcag gaggctttcg catgagtatt
     2461 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     2521 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc gcgagtgggt
     2581 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgc
     2641 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     2701 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtat
     2761 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     2821 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat tggaggaccg
     2881 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     2941 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     3001 atggcaacaa ccttgcgtaa actattaact ggcgaactac ttactctagc ttcccggcaa
     3061 cagttgatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     3121 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     3181 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     3241 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     3301 aagcattggt aaccgattct aggtgcattg gcgcagaaaa aaatgcctga tgcgacgctg
     3361 cgcgtcttat actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc
     3421 cacttccata cgtgtcctcc ttaccagaaa tttatcctta agatcccgaa tcgtttaaac
     3481 tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta cgcgaacagc cgtggcgctc
     3541 atttgctcgt cgggcatcga atctcgtcag ctatcgtcag cttacctttt tggcagcgat
     3601 cgcggctccc gacatcttgg accattagct ccacaggtat cttcttccct ctagtggtca
     3661 taacagcagc ttcagctacc tctcaattca aaaaacccct caagacccgt ttagaggccc
     3721 caaggggtta tgctatcaat cgttgcgtta cacacacaaa aaaccaacac acatccatct
     3781 tcgatggata gcgattttat tatctaactg ctgatcgagt gtagccctta accgcttttg
     3841 aatgtccgtc ttagttaatc cctaattgaa cgctccaaaa ggtgacttcc aatagggatt
     3901 tattcctttt aaaattaacg gcataatcgt agatcgcatg ccgggaggtg gtccctgcag
     3961 ttacgccaat gataaccccc gccagaaaaa tcttagtagc cttccctttt tgttttccgt
     4021 gccccaactc ggcggattga ctcggcccct tccggaaaca cccgaatcaa cttctagtca
     4081 aattattgtt cacgccgcaa tgacccaccc ctggcccgcg tctgtggaac tgacccctgg
     4141 tgtacaggag agttcgctgc tgaaagtggt cccaaagggg tactagtttt taagctccca
     4201 actccccctc ccccagcgtc tggaggattc cacaccctcg caccgcaggg gcgaggaagt
     4261 gggcggagtc cggttttggc gccagccgct gaggctgcca agcagaaaag ccaccgctga
     4321 ggagactccg gtcactgtcc tcgccccgcc tcccccttcc ctccccttgg ggaccaccgg
     4381 gcgccacgcc gcgaacggta agtgccgcgg tcgtcggcgc ctccgccctc cccctagggc
     4441 cccaattccc agcgggcgcg gcgcgcggcc cctccccccg ccgggcgcgc gcccgctgcc
     4501 ccgcccttcg tggccgcccg gcgtgggcgg tgccacccct ccccccggcg gccccgcgcg
     4561 cagctcccgg ctccctcccc cttcggatgt ggcttgagct gtaggcgcgg aggggctagc
     4621 tcgtgaggct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag
     4681 ttggggggag gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg
     4741 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata
     4801 agtgcactag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggta
     4861 agtgccgtgt gtggttcccg cgggcctggc ctctttacgg gttatggccc ttgcgtgcct
     4921 tgaattactt ccacctggct gcagtacgtg attcttgatc ccgagcttcg ggttggaagt
     4981 gggtgggaga gttcgtggcc ttgcgcttaa ggagcccctt cgcctcgtgc ttgagttgtg
     5041 gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc
     5101 gctgctttcg ataagtctct agccatttaa aatttttgat gacctgctgc gacgcttttt
     5161 ttctggcaag atagtcttgt aaatgcgggc caagatcagc acactggtat ttcggttttt
     5221 ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca catgttcggc gaggcggggc
     5281 ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc aagctgcccg gcctgctctg
     5341 gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg
     5401 gcaccagttg cgtgagcgga aagatggccg cttcccggcc ctgctgcagg gagcacaaaa
     5461 tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac ccacacaaag gaaaagggcc
     5521 tttccgtcct cagccgtcgc ttcatgtgac tccacggagt accgggcgcc gtccaggcac
     5581 ctcgattagt tctccagctt ttggagtacg tcgtctttag gttgggggga ggggttttat
     5641 gcgatggagt ttccccacac tgagtgggtg gagactgaag ttaggccagc ttggcacttg
     5701 atgtaattct ccttggaatt tgcccttttt gagtttggat cttggttcat tctcaagcct
     5761 cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt cgtgaaaact acccctaaaa
     5821 gccaaaagat ctttgtcgat cctaccatcc actcgacaca cccgccagcg gccgctgcca
     5881 agcttgctag gaattccgcg tctgcaggat atcaagcttg cggtacgtga gtctatggga
     5941 cgcttgatgt tttctttccc cttcttttct atggttaagt tcatgtcata ggaaggggat
     6001 aagtaacagg gtacagttta gaatgggaaa cagacgaatg attgcatcag tgtggaagtc
     6061 tcaggatcgt tttagtttct tttatttgct gttcataaca attgttttct tttgtttaat
     6121 tcttgctttc tttttttttc ttctccgcaa tttttactat tatacttaat gccttaacat
     6181 tgtgtataac aaaaggaaat atctctgaga tacattaagt aacttaaaaa aaaactttac
     6241 acagtctgcc tagtacatta ctatttggaa tatatgtgtg cttatttgca tattcataat
     6301 ctccctactt tattttcttt tatttttaat tgatacataa tcattataca tatttatggg
     6361 ttaaagtgta atgttttaat atgtgtacac atattgacca aatcagggta attttgcatt
     6421 tgtaatttta aaaaatgctt tcttctttta atatactttt ttgtttatct tatttctaat
     6481 actttcccta atctctttct ttcagggcaa taatgataca atgtatcatg cctctttgca
     6541 ccattctaaa gaataacagt gataatttct gggttaaggc aatagcaata tctctgcata
     6601 taaatatttc tgcatataaa ttgtaactga tgtaagaggt ttcatattgc taatagcagc
     6661 tacaatccag ctaccattct gcttttattt tatggttggg ataaggctgg attattctga
     6721 gtccaagcta ggcccttttg ctaatcatgt tcatacctct tatcttcctc ccacagcgcg
     6781 ggcccggcca ccatggcccc aaagaagaag cggaaggtcg gtatccacgg agtcccagca
     6841 gccacacagt tcgagggctt taccaacctg tatcaggtga gcaagacact gcggtttgag
     6901 ctgatcccac agggcaagac cctgaagcac atccaggagc agggcttcat cgaggaggac
     6961 aaggcccgca atgatcacta caaggagctg aagcccatca tcgatcggat ctacaagacc
     7021 tatgccgacc agtgcctgca gctggtgcag ctggattggg agaacctgag cgccgccatc
     7081 gactcctata gaaaggagaa aaccgaggag acaaggaacg ccctgatcga ggagcaggcc
     7141 acatatcgca atgccatcca cgactacttc atcggccgga cagacaacct gaccgatgcc
     7201 atcaataaga gacacgccga gatctacaag ggcctgttca aggccgagct gtttaatggc
     7261 aaggtgctga agcagctggg caccgtgacc acaaccgagc acgagaacgc cctgctgcgg
     7321 agcttcgaca agtttacaac ctacttctcc ggcttttatc ggaacaggaa gaacgtgttc
     7381 agcgccgagg atatcagcac agccatccca caccgcatcg tgcaggacaa cttccccaag
     7441 tttaaggaga attgtcacat cttcacacgc ctgatcaccg ccgtgcccag cctgcgggag
     7501 cactttgaga acgtgaagaa ggccatcggc atcttcgtga gcacctccat cgaggaggtg
     7561 ttttccttcc ctttttataa ccagctgctg acacagaccc agatcgacct gtataaccag
     7621 ctgctgggag gaatctctcg ggaggcaggc accgagaaga tcaagggcct gaacgaggtg
     7681 ctgaatctgg ccatccagaa gaatgatgag acagcccaca tcatcgcctc cctgccacac
     7741 agattcatcc ccctgtttaa gcagatcctg tccgatagga acaccctgtc tttcatcctg
     7801 gaggagttta agagcgacga ggaagtgatc cagtccttct gcaagtacaa gacactgctg
     7861 agaaacgaga acgtgctgga gacagccgag gccctgttta acgagctgaa cagcatcgac
     7921 ctgacacaca tcttcatcag ccacaagaag ctggagacaa tcagcagcgc cctgtgcgac
     7981 cactgggata cactgaggaa tgccctgtat gagcggagaa tctccgagct gacaggcaag
     8041 atcaccaagt ctgccaagga gaaggtgcag cgcagcctga agcacgagga tatcaacctg
     8101 caggagatca tctctgccgc aggcaaggag ctgagcgagg ccttcaagca gaaaaccagc
     8161 gagatcctgt cccacgcaca cgccgccctg gatcagccac tgcctacaac cctgaagaag
     8221 caggaggaga aggagatcct gaagtctcag ctggacagcc tgctgggcct gtaccacctg
     8281 ctggactggt ttgccgtgga tgagtccaac gaggtggacc ccgagttctc tgcccggctg
     8341 accggcatca agctggagat ggagccttct ctgagcttct acaacaaggc cagaaattat
     8401 gccaccaaga agccctactc cgtggagaag ttcaagctga actttcagat gcctacactg
     8461 gcccgcggct gggacgtgaa tcgggagaag aacaatggcg ccatcctgtt tgtgaagaac
     8521 ggcctgtact atctgggcat catgccaaag cagaagggca ggtataaggc cctgagcttc
     8581 gagcccacag agaaaaccag cgagggcttt gataagatgt actatgacta cttccctgat
     8641 gccgccaaga tgatcccaaa gtgcagcacc cagctgaagg ccgtgacagc ccactttcag
     8701 acccacacaa cccccatcct gctgtccaac aatttcatcg agcctctgga gatcacaaag
     8761 gagatctacg acctgaacaa tcctgagaag gagccaaaga agtttcagac agcctacgcc
     8821 aagaaaaccg gcgaccagaa gggctacaga gaggccctgt gcaagtggat cgacttcaca
     8881 agggattttc tgtccaagta taccaagaca acctctatcg atctgtctag cctgcggcca
     8941 tcctctcagt ataaggacct gggcgagtac tatgccgagc tgaatcccct gctgtaccac
     9001 atcagcttcc agagaatcgc cgagaaggag atcatggatg ccgtggagac aggcaagctg
     9061 tacctgttcc agatctataa caaggacttt gccaagggcc accacggcaa gcctaatctg
     9121 cacacactgt attggaccgg cctgttttct ccagagaacc tggccaagac aagcatcaag
     9181 ctgaatggcc aggccgagct gttctaccgc cctaagtcca ggatgaagag gatggcacac
     9241 cggctgggag agaagatgct gaacaagaag ctgaaggatc agaaaacccc aatccccgac
     9301 accctgtacc aggagctgta cgactatgtg aatcacagac tgtcccacga cctgtctgat
     9361 gaggccaggg ccctgctgcc caacgtgatc accaaggagg tgtctcacga gatcatcaag
     9421 gataggcgct ttaccagcga caagttcttt ttccacgtgc ctatcacact gaactatcag
     9481 gccgccaatt ccccatctaa gttcaaccag agggtgaatg cctacctgaa ggagcacccc
     9541 gagacaccta tcatcggcat cgatcggggc gagagaaacc tgatctatat cacagtgatc
     9601 gactccaccg gcaagatcct ggagcagcgg agcctgaaca ccatccagca gtttgattac
     9661 cagaagaagc tggacaacag ggagaaggag agggtggcag caaggcaggc ctggtctgtg
     9721 gtgggcacaa tcaaggatct gaagcagggc tatctgagcc aggtcatcca cgagatcgtg
     9781 gacctgatga tccactacca ggccgtggtg gtgctggaga acctgaattt cggctttaag
     9841 agcaagagga ccggcatcgc cgagaaggcc gtgtaccagc agttcgagaa gatgctgatc
     9901 gataagctga attgcctggt gctgaaggac tatccagcag agaaagtggg aggcgtgctg
     9961 aacccatacc agctgacaga ccagttcacc tcctttgcca agatgggcac ccagtctggc
    10021 ttcctgtttt acgtgcctgc cccatataca tctaagatcg atcccctgac cggcttcgtg
    10081 gaccccttcg tgtggaaaac catcaagaat cacgagagcc gcaagcactt cctggagggc
    10141 ttcgactttc tgcactacga cgtgaaaacc ggcgacttca tcctgcactt taagatgaac
    10201 agaaatctgt ccttccagag gggcctgccc ggctttatgc ctgcatggga tatcgtgttc
    10261 gagaagaacg agacacagtt tgacgccaag ggcacccctt tcatcgccgg caagagaatc
    10321 gtgccagtga tcgagaatca cagattcacc ggcagatacc gggacctgta tcctgccaac
    10381 gagctgatcg ccctgctgga ggagaagggc atcgtgttca gggatggctc caacatcctg
    10441 ccaaagctgc tggagaatga cgattctcac gccatcgaca ccatggtggc cctgatccgc
    10501 agcgtgctgc agatgcggaa ctccaatgcc gccacaggcg aggactatat caacagcccc
    10561 gtgcgcgatc tgaatggcgt gtgcttcgac tcccggtttc agaacccaga gtggcccatg
    10621 gacgccgatg ccaatggcgc ctaccacatc gccctgaagg gccagctgct gctgaatcac
    10681 ctgaaggaga gcaaggatct gaagctgcag aacggcatct ccaatcagga ctggctggcc
    10741 tacatccagg agctgcgcaa caaaaggccg gcggccacga aaaaggccgg ccaggcaaaa
    10801 aagaaaaagg gcagcagcgg atccagcgag ctgattaagg agaacatgca catgaagctg
    10861 tacatggagg gcaccgtgga caaccatcac ttcaagtgca catccgaggg cgaaggcaag
    10921 ccctacgagg gcacccagac catgagaatc aaggtggtcg agggcggccc tctccccttc
    10981 gccttcgaca tcctggctac tagcttcctc tacggcagca agaccttcat caaccacacc
    11041 cagggcatcc ccgacttctt caagcagtcc ttccctgagg gcttcacatg ggagagagtc
    11101 accacatacg aagacggggg cgtgctgacc gctacccagg acaccagcct ccaggacggc
    11161 tgcctcatct acaacgtcaa gatcagaggg gtgaacttca catccaacgg ccctgtgatg
    11221 cagaagaaaa cactcggctg ggaggccttc accgagacgc tgtaccccgc tgacggcggc
    11281 ctggaaggca gaaacgacat ggccctgaag ctcgtgggcg ggagccatct gatcgcaaac
    11341 atcaagacca catatagatc caagaaaccc gctaagaacc tcaagatgcc tggcgtctac
    11401 tatgtggact acagactgga aagaatcaag gaggccaaca acgagaccta cgtcgagcag
    11461 cacgaggtgg cagtggccag atactgcgac ctccctagca aactggggca caagcttaat
    11521 tagtaactag cttgactgac tgagatacag cgtaccttca gctcacagac atgataagat
    11581 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
    11641 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
    11701 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa
    11761 gcaagtaaaa cctctacaaa tgtggtattg gcccatctct atcggtatcg tagcataacc
    11821 ccttggggcc tctaaacggg tcttgagggg ttttttgtgc ccctcgggcc ggattgctat
    11881 ctaccggcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac
    11941 atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct
    12001 ccttaccaga aatttatcct taaggtcgtc agctatcctg caggcgatct ctcgatttcg
    12061 atcaagacat tcctttaatg gtcttttctg gacaccacta ggggtcagaa gtagttcatc
    12121 aaactttctt ccctccctaa tctcattggt taccttgggc tatcgaaact taattaacca
    12181 gtcaagtcag ctacttggcg agatcgactt gtctgggttt cgactacgct cagaattgcg
    12241 tcagtcaagt tcgatctggt ccttgctatt gcacccgttc tccgattacg agtttcattt
    12301 aaatcatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
    12361 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
    12421 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
    12481 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
    12541 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
    12601 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
    12661 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
    12721 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
    12781 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
    12841 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
    12901 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caa
//
