LOCUS       sequence-455504-       16012 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of beta actin driven BFP and molecular barcode array for 
            SABER-seq in zebrafish. Also contains heat-shock activated mCherry 
            reporter and CreERT2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16012)
  AUTHORS   Siniscalco AM, Perera RP, Greenslade JE, Veeravenkatasubramanian H, 
            Masters A, Doll HM, Raj B
  TITLE     Barcoding Notch signaling in the developing brain.
  JOURNAL   Development. 2024 Nov 22:dev.203102. doi: 10.1242/dev.203102.
  PUBMED    39575683
REFERENCE   2  (bases 1 to 16012)
  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: 
            "Development. 2024 Nov 22:dev.203102. doi: 10.1242/dev.203102."
COMMENT     Sequence Label: p140Tol2-Olactb:SABER
FEATURES             Location/Qualifiers
     source          1..16012
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(34..462)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(144..165)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     356..375
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     589..611
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     603..620
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     604..620
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        630..648
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    join(2499..2510,2511..2673)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     CDS             join(7144..7146,7147..7149,7150..7863)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7450..7469)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7797..7818
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    8301..8435
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    8644..8778
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(8851..9744)
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPVKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATA"
     CDS             complement(8857..9792)
                     /codon_start=1
                     /product="mutated ligand-binding domain of the human 
                     estrogen receptor (Feil et al., 1997)"
                     /label=ERT2
                     /translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
                     YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
                     LMIGLVWRSMEHPVKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
                     VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
                     AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEET
                     DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
     CDS             complement(9805..10830)
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="SNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLL
                     SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP
                     RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL
                     GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER
                     WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR
                     YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED
                     GD"
     CDS             complement(10837..10890)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             complement(10891..11598)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(11166..11185)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     11435..11453
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      11595..11604
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(11625..11649)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    join(13542..13729,13730..13741)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(13775..13791)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13775..13791)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(13828..13848)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(13828..13846)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(13867..13883)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13867..13883)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(13880..13902)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    13891..13907
                     /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(13915..13921,13922..13939,13940..13945))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13960..13981
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(14098..14115)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(14269..14857)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(14349..14368)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(15028..15819,15820..15888))
                     /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     15651..15670
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(15889..15993)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
       61 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
      121 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
      181 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
      241 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
      301 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
      361 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
      421 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc
      481 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
      541 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
      601 gttgtaaaac gacggccagt gagcgcgcgt aatacgactc actatagggc gaattgggta
      661 ccaaatagta ggaattaccc acctgtacaa gtgctgaaaa cttggatgaa taagcccgtt
      721 tgccattttc tactgctatt ttaaatcttt tctgtatttg tctgcatttg tctttaccct
      781 tgaataaatg ttttagttgt tttttttcaa ttatgactgt gtttaacaga cattattaca
      841 ataatatgta atagtagtac acactattat tatgtaatag tacttgttga ctgtatttga
      901 gaactggacc gagtgagtgt tacgtcaccc attcaaaatg acttacttct ggctccaaca
      961 aaatgaagtt aattcagttg ccatttttca ctgtatggac atcgccgtgt tggagctaga
     1021 cgttgccaat aagcaagaaa gagccgagat gcgtcgagtc tgagtcaccg ttcctatggc
     1081 aacccctcta accaatcaga agtaagcttg ttggaagtcc acagcctacc acttgaaagc
     1141 gggctgcaca aaatctgtca aacgttttga acgttggatg tgagagcaca tacttttatt
     1201 aaggcatctg gttggtcagt ttataatatc aacaacttgg gctacagaaa agaaaagtta
     1261 ttacagaaat tatgattaac aagtacatgt taaataaaga ttttaatatg aatgccacca
     1321 ctggagcatt catgccattt ggagcttctt cctgtttgga tcactagaag gaggaggtca
     1381 ctcattacag ttctcatata cagtcgttgg ttggttggtt ggttggtaga ttgattgatt
     1441 gattgattga ttgattgatt gattgattga ttgattgatt gattgattga ttgattgatt
     1501 gattggtagt caaaataaga aataatttcc acagattcat tacagaaatg attaaatgca
     1561 tacataaaaa actggggggg ggggggatac aacaacacac ttaagtcaca tttgcctacg
     1621 taaagaaaag taaagaaaat caatagctat attttacatc tccttttttt gctgtctttt
     1681 aattagcctt gttttgctgt tatcttgatt gaaactgtaa cttcttcacc tgcttctttt
     1741 ctttgtaggt tttgccagcc cagaaacagg catggctgtg caaggccaga gcaccatgca
     1801 caatgcgctg catgtcttca tgaacggctc aatgtcctca gtccagggct cagccaacga
     1861 ccccatcttc ctccttcacc atgctttcat tgacaggtaa caaacacgtc atgacattag
     1921 actgcacagt ttttgacaaa gttcatacaa tctgttgttt atagctgcta caattagtga
     1981 agtttgtgaa tgtacttgga tgagcagcga aagatcaatt gagatcaatt gttagagttt
     2041 ggttgccctg cagagcaaag aacaaaaaat aatctggtgg ctttactgcg tgaggttatt
     2101 attggtggaa tagaaacaca aaacataatt gcatttattt gtttaatttt ttatcttatc
     2161 ttaactttca tcttgcatat ttgtttctta catcatttct agcatctttg agcgctggct
     2221 aagaactcat cagcctcccc ggtccatcta cccacgtacc aatgcaccaa ttggccacaa
     2281 tgacggctac tacatggtgc cattccttcc tctttatagg aatggagact acctcctgtc
     2341 caacaatgct cttggatacg agtacgccta cctgttggac ccaggtcatt gcacaacacc
     2401 agaaatgccc tctgatctgc aaaagacgtg aatatctgtt cagacaccca tatccactct
     2461 gttccacaca ggtcagaggt ttgtccagga gttcttgaca gaggtgtaaa aagtactcaa
     2521 aaattttact caagtgaaag tacaagtact tagggaaaat tttactcaat taaaagtaaa
     2581 agtatctggc tagaatctta cttgagtaaa agtaaaaaag tactccatta aaattgtact
     2641 tgagtattaa ggaagtaaaa gtaaaagcaa gaaagaaaac tagagattct tgtttaagct
     2701 tttaatctca aaaaacatta aatgaaatgc atacaaggtt ttatcctgct ttagaactgt
     2761 ttgtatttaa ttatcaaact ataagacaga caatctaatg ccagtacacg ctactcaaag
     2821 ttgtaaaacc tcagatttaa cttcagtaga agctgattct caaaattgtt agtgtcaagc
     2881 ctagctcttt tggggctgaa aagcaatcct gcagtgctga aaagcctctc acaggcagcc
     2941 gatgcgggaa gaggtgtatt agtcttgata gagaggctgc aaatagcagg aaacgtgagc
     3001 agagactccc tggtgtctga aacacaggcc agatgggccc tcgagcaggg ggatcatcta
     3061 atcaagcaca aataaggggc gtgtaacaca aaagccagcg accctttcca atgcaaatca
     3121 aacttgcaat tctttgccgt ttttatcatt taagtgtcgg cttaaggtcc actatcagat
     3181 gtaaacagcc ttatctaaca aaggtatcat tacattctga aattctcagg catgcaagct
     3241 agcttatgac gcactaggga gtgccaccct tcctttcgcc ctaacttcgt gataactcgc
     3301 gcgtttcact caacagctgc atccgcccta gtgctactgg gagttgtagt atacaagacg
     3361 cttacaggct gaatgttctg tcaagacccc gcctctagca ctttgggaat tcaattacag
     3421 tggtcatgaa tattatatct aaccacagag ctaaagaaaa aacataccaa gacagttttt
     3481 tttctgttga tgggtggtca ttctacattg aagtgaatgg ggtctgactg gtttgtttta
     3541 cgtgtcagct agtgctttct tggtgaactg atctggctca tttttttact ctgacctcat
     3601 ctgagaattg ttcaaaacaa atgcacgata tttaatactg ctgataagaa ggattcgctt
     3661 cacttgcagg tgaaccctgg tgtggcaata gtgttaatga aaacaaacta ctcggcagta
     3721 aaagtatgtg acatgaataa aaaaaaatca caaactggaa gaagatcaga tcatgaaaga
     3781 attgtaagaa tgtcccattg tacgatgtgt tgtggtaaag tggtgaagtc acttatcaat
     3841 ccatgatggt cagagaacta gcaggactgg ttttcttttc tttgtttttt tttatttcct
     3901 tcatcctggt actgcccact tctggccttc aaacaaagtc aagtattttt tttattcaaa
     3961 agcagatcaa atttagaagg taacatcatc tgcccatttg aatcaagtct gtagaactat
     4021 cttagtatga acttgccata atgcttcata agaagtgatt gcaaatcacc taccacacag
     4081 ctggatgagc tggaactctg acacatacat tcattcacct gcaatcacgt aaggagccta
     4141 ctacgaaggg acactcctgt gaggcaggat tacatccttt cccagccaac ccctgtctgt
     4201 gtttttccag tttcagcagt cagtccttcc agtgtgctct taactttacc agaacccgcc
     4261 ctcccagaat actgacaaat agaaataaaa atgtgtttta gatggacgca ccttgaatgg
     4321 gttacagatt tcatttcagg cctgtaaata agtaaatgca gtgagttggc ctttgatcga
     4381 caagaacgat tcattccata tgagagtgtt ttttatgaga gatttaaaac tcttacataa
     4441 agaattattt acatacaact ctcgagagag aactgtttca ccccacaagc agaagctttt
     4501 atttcagcgg gttaataaaa caaaacaaaa atggacattt aaaagttagg tctgctgatc
     4561 agaaatgcaa gaatctgcta attcagaagg aaatgaagca tgttttgttt gaaacttgat
     4621 taaaaagctg aaacgtttcc atgtataaag tggaaatgtg ttgatgaaaa taaagaccat
     4681 ttaaaactta aagccacgaa tgaatttaag gaagaggaga tggcatttag cataaaagaa
     4741 ttggtgtaac tgagagcgag gaagaggaag tggtacaaga ggaagaaaag tagttgctgc
     4801 ttgtgtctcg cagacagtga catgtttaaa tggcctctgg gttatttaac ttgtatgggc
     4861 agctatgttt ctggagcaaa aaaagtttaa ataaactgtg aaactaatga atggaagaca
     4921 agtaaggtaa aagcaaagaa ccctaagcct ataaaaccca aagtcaggaa agttacaccc
     4981 tttgcagtaa accacagaga ggaaataaga gcatgtgtga ctttatttgt ctctctctga
     5041 gtttaatcga ttaaggtgaa tgtatagtag cgtatgggtt gggtctgaca gtggttattt
     5101 tcactcaata ggtcatttat gactaaaaga agatgcaagt cagccttaat gtcctcttat
     5161 gtggaccttt tttcttcatt tgtaggggaa agaggaagta gcaagagctc tgagaagtta
     5221 acaagaatcc agactaatgc catacatggt caattggtat tagtgagatt gagtttttac
     5281 cattgcttag tgggaaaagt ggttattgtc catcctctga cagggcactt ttacagtcca
     5341 tagtgtcttt tgtgagtcat gtgtttggct tggagggtct gtgactccat ctgaggtggc
     5401 attctgcttt ccagagtgta cagcaacatc cacgttgtaa ttgtcctcaa attgcatcaa
     5461 acatatcttg catttccaga ctgaaatctt gagagacagg ttttaagagg aaaagtattc
     5521 acagggagga gcagcctgtg ctctgctgtt tgaatgatca gataagcagg gggacttgct
     5581 cgacagcttt atacaggttc atattttata tttatttgaa actttgtaaa ctctatatgt
     5641 attaatattt gttcaagaaa cattatatcc ttagattagg cctcacatca atccctatcc
     5701 ttttattttc ccctagcccg ttactgttat cgcctccctc tttctacgct cagtgcacca
     5761 cagcgtgcac ccgacgtgcc ccagtgagtg acgctggacc aatcacatgc cgcgattccg
     5821 aaagtttacc ttttatggaa agagccgggc aacggacgga ctataaatac cacgtgccca
     5881 cggctagcaa attcactctg agcgccgtca cacacagctt gtgcgggata tcattcgcct
     5941 gaaaccgatt cccttgaaac gaaaagcccc ccacccaaag gtaagataga gacgatattt
     6001 ggtttaattc aaatgttttg gaaaccttac gattgtttaa ttgtgcagtt taattcttaa
     6061 ttggaggtga ccattagcag aattttgtgg ccactaaatt tatttttagg cctgaaatgg
     6121 ttaaactttt aattatcttt ttaaaattga caattttgtt tgttttggca tcaatcaagt
     6181 tgtaccaaaa ttagccactt aaacgcacag gtgccaattg attgtaaccg gcaaatggct
     6241 tttcagtttg gttttaattg tatattttat ataggcaact aaataattac tattcgtatc
     6301 ttacatggtg gggtgaattt ttcgagtcct tctacggacg ttacccgtcc gagttgaggg
     6361 tgtctggatc ctggcgggtc cctttgtgcg ccgttacggg gtgtgaccta ctttagcata
     6421 ttagcctagc ttcatgctaa cacgcccttc tagacttgac tctgaatcct cacatgtagg
     6481 ggggtgggac tgttatacat ttgtttgtat aaaatgtaat aacccttaaa gcattttaaa
     6541 tgtattcgat taaattgacc atgtaactgt agtttgacgg cgaagaaaag ggcaaatgac
     6601 aaatcattca ttcaagaaaa ggcgtggctt tcaggaaatg attgatcagt catgggctcg
     6661 cacacgtttt aatttcctac taaagcttga cgggtcgtgg tgctttatgc cagcaacgac
     6721 ttcgcacaaa ttatgacgca gtgattaaga aatttttcag tcttgtgtaa caatgggagg
     6781 gaacggggac aaagtagagc tgtaatgaac cagtgggagt aggctttcgt cgccgggtcc
     6841 gatagtctaa agtccctccc ctcttgctgt actttggccg cgtgccttgc cggtagccgc
     6901 aaagctgctc aaaaccgaac tatgtcctta tatggtaaca agagaatgca gcaccacttc
     6961 cttttgtctg ggctcggaat gtagcgctct acctccacca agcggacact ttggggaact
     7021 gccgctccct gacaggaaat gggacatctg actgcaaagc agatgcagat gggcgtgtct
     7081 ttggtgttaa cgcctttttt gtttgttctt ttccagttta gccgtcgaca ccggtctgac
     7141 aacatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     7201 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     7261 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     7321 accctcgtga ccaccctgac ctggggcgtg cagtgcttcg cccgctaccc cgaccacatg
     7381 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     7441 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     7501 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     7561 cacaagctgg agtacaacgc catcagcgac aacgtctata tcaccgccga caagcagaag
     7621 aacggcatca aggccaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     7681 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     7741 cactacctga gcacccagtc caagctgagc aaagacccca acgagaagcg cgatcacatg
     7801 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     7861 taaagcggcg tacgctcgag ctcaagcttc gtcgtagatc ttcatcgtga tgcgtttaca
     7921 accaaacctg ccagtgcgcc gcaatttatt cgtgtggatg cattattacg tgtctttgcg
     7981 catgagtcgg cggcgtttct gcctgtgcca actgcgcaga gagacgcgag tctgacatga
     8041 tgatgaagag tcgaggagag tgagccagtg cgccggaaac tacatggggt cagctcgtct
     8101 ctccagcagt tcccccgagt cgtgccctga agctgaagga cggcggccac ggagttcaag
     8161 tccatctaca tggccaagct gcccggctac tactacgtgg acgcggactg gaggacttct
     8221 ggggaggtgc tcgacctcga gacaaatggc agcccggggg atccaaactc gagaaagcta
     8281 gcaaaccatg gtacgatgat gatccagaca tgataagata cattgatgag tttggacaaa
     8341 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     8401 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     8461 tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat
     8521 gtggtatggc tgattatgat cctctagatc agtggccgga cgcgtatggc gcgccgcgaa
     8581 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     8641 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     8701 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     8761 tcttatcatg tctggatcta cgtaatacga ctcactatag ttctagaggc tcgagaggcc
     8821 ttgaattccg ccagggagct cagatcttca tcaagctgtg gcagggaaac cctctgcctc
     8881 ccccgtgatg taatactttt gcaaggaatg cgatgaagta gagcccgcag tggccaagtg
     8941 gctttggtcc gtctcctcca cggatgcccc tccacggcta gtgggcgcat gtaggcggtg
     9001 ggcgtccgcc gcctccagca gcaggtcata gaggggcacc acgttcttgc acttcatgct
     9061 gtacagatgc tccatgcctt tgttactcat gtgcctgatg tgggagagga tgaggaggag
     9121 ctgggccagc cgctggtgct gctgctgcag ggtcaggcct gccttggcca tcaggtggat
     9181 caaagtgtct gtgatcttgt ccaggactcg gtggatatgg tccttctctt ccagagactt
     9241 cagggtgctg gacagaaatg tgtacactcc agaattaagc aaaataatag atttgaggca
     9301 cacaaactcc tctccctgca gattcatcat gcggaaccga gatgatgtag ccagcagcat
     9361 gtcgaagatc tccaccatgc cctctacaca ttttccctgg ttcctgtcca agagcaagtt
     9421 aggagcaaac agtagcttca ctgggtgctc catggagcgc cagacgagac caatcatcag
     9481 gatctctagc caggcacatt ctagaaggtg gacctgatca tggagggtca aatccacaaa
     9541 gcctggcacc ctcttcgccc agttgatcat gtgaaccagc tccctgtctg ccaggttggt
     9601 cagtaagccc atcatcgaag cttcactgaa gggtctggta ggatcatact cggaatagag
     9661 tatggggggc tcagcatcca acaaggcact gaccatctgg tcggccgtca gggacaaggc
     9721 caggctgttc ttcttagagc gtttgatcat gagcgggctt ggccaaaggt tggcagctct
     9781 catgtctcca gcagatggct cgagatcgcc atcttccagc aggcgcacca ttgcccctgt
     9841 ttcactatcc aggttacgga tatagttcat gacaatattt acattggtcc agccaccagc
     9901 ttgcatgatc tccggtattg aaactccagc gcgggccata tctcgcgcgg ctccgacacg
     9961 ggcactgtgt ccagaccagg ccaggtatct ctgaccagag tcatccttag cgccgtaaat
    10021 caatcgatga gttgcttcaa aaatcccttc cagggcgcga gttgatagct ggctggtggc
    10081 agatggcgcg gcaacaccat tttttctgac ccggcaaaac aggtagttat tcggatcatc
    10141 agctacacca gagacggaaa tccatcgctc gaccagttta gttaccccca ggctaagtgc
    10201 cttctctaca cctgcggtgc taaccagcgt tttcgttctg ccaatatgga ttaacattct
    10261 cccaccgtca gtacgtgaga tatctttaac cctgatcctg gcaatttcgg ctatacgtaa
    10321 cagggtgtta taagcaatcc ccagaaatgc cagattacgt atatcctggc agcgatcgct
    10381 attttccatg agtgaacgaa cctggtcgaa atcagtgcgt tcgaacgcta gagcctgttt
    10441 tgcacgttca ccggcatcaa cgttttcttt tcggatccgc cgcataacca gtgaaacagc
    10501 attgctgtca cttggtcgtg gcagcccgga ccgacgatga agcatgttta gctggcccaa
    10561 atgttgctgg atagttttta ctgccagacc gcgcgcctga agatatagaa gataatcgcg
    10621 aacatcttca ggttctgcgg gaaaccattt ccggttattc aacttgcacc atgccgccca
    10681 cgaccggcaa acggacagaa gcattttcca ggtatgctca gaaaacgcct ggcgatccct
    10741 gaacatgtcc atcaggttct tgcgaacctc atcactcgtt gcatcgaccg gtaatgcagg
    10801 caaattttgg tgtacggtca gtaaattgga gctagcaggg ccgggattct cctccacgtc
    10861 accgcatgtt agaagacttc ctctgccctc cttgtacagc tcgtccatgc cgccggtgga
    10921 gtggcggccc tcggcgcgtt cgtactgttc cacgatggtg tagtcctcgt tgtgggaggt
    10981 gatgtccaac ttgatgttga cgttgtaggc gccgggcagc tgcacgggct tcttggcctt
    11041 gtaggtggtc ttgacctcag cgtcgtagtg gccgccgtcc ttcagcttca gcctctgctt
    11101 gatctcgccc ttcagggcgc cgtcctcggg gtacatccgc tcggaggagg cctcccagcc
    11161 catggtcttc ttctgcatta cggggccgtc ggaggggaag ttggtgccgc gcagcttcac
    11221 cttgtagatg aactcgccgt cctgcaggga ggagtcctgg gtcacggtca ccacgccgcc
    11281 gtcctcgaag ttcatcacgc gctcccactt gaagccctcg gggaaggaca gcttcaagta
    11341 gtcggggatg tcggcggggt gcttcacgta ggccttggag ccgtacatga actgagggga
    11401 caggatgtcc caggcgaagg gcagggggcc acccttggtc accttcagct tggcggtctg
    11461 ggtgccctcg taggggcggc cctcgccctc gccctcgatc tcgaactcgt ggccgttcac
    11521 ggagccctcc atgtgcacct tgaagcgcat gaactccttg atgatggcca tgttatcctc
    11581 ctcgcccttg ctcaccatgg tggccggccc atatggcccg ggccgttcaa cttttttgta
    11641 caaacttggc aggaaaaaaa aacaattaga attaatttta tatttataca atttatggtg
    11701 caattgtttc attatgaaag gatttttcta gtcaacaagt gaaattctct taataaccaa
    11761 gttattatta gttgtacaag taaaacgcta attttctttt gtagacaaca ttaacacaaa
    11821 ttcatttagg acatttgcta gtatttatgg ttattatgct catataaaat aatgtcagca
    11881 acataaagta aatgcccgtc tgttcattgc tttttttaaa aatccttatt gtgaaaacgg
    11941 gaaatataat tttctagtag acctaaaata agatgaacat aggcagtcca catgtaaaaa
    12001 gattacatat acaaagttga ttttttgaca ttaagtacat ataatctcta tataattctg
    12061 attagccaat agagtacaat atattgagac ttaccttaca gtagattctt aaatagagcg
    12121 aataaacacc tcgtcgggga aaaagtcctg tcaggctgct cttttcttct ttgtcaagtc
    12181 gcttctcttc ggttgagttt ctctgcggtc tctctttcgc cctttaaata cagcagagcg
    12241 cgcacccgtt cagactctac actgttctgg aagcctccag tgaattcgac caatcaaaac
    12301 acagcaggca gccgcgtcac ttcttcagct gtcattgaga ggagccgctg agaggaatgt
    12361 agtggagatt tccagagttt taccgtgaaa ttacgcgaaa cagcggtcaa aaatacaaag
    12421 agatttaaaa tgtttttttt atttgagtgt tttattgtac ttattgctga aatatgtgcg
    12481 gttgtgtgat gtgttgtttt tccagttgca atgaatgaac aaactaaaat gataaaatgt
    12541 ttgaacgatt cagtaaaaaa aaatgcaata aaaaaatctt gctgaaatgt tttacacagt
    12601 agcaaaatcg ggcctgtggt gttttcagct acatagaagg caaacctgtt ttatataagc
    12661 taatagcttt ggtcagattt gtttaacgtt tcaacaaaac aaatggatgt ttactgtact
    12721 atattgattg gttaaaacca atactttatt ctgggattgc agtcttaact atttttgtag
    12781 tgttaaaacc aacttgctaa cttgagaggt catgtttacc ctcagagaaa taggcacaag
    12841 tctctttgtt taatattgga gatcatcctc cttgttcagt cgtggtctgt atttaatttt
    12901 aatgtatatg attgccgtaa aagtgttaga aagtttaccc tggtgctatg aaatcaatct
    12961 gctgcttttg acccagttag tgtgtcttta atctaactta atgagacccc agtgtggtca
    13021 acaatcaata gaaacaaagt gtgttgtttt ataactgtta actcctatta ttttttatct
    13081 tctgtagcat atagacaaaa catgctcgtt ctaatagttt aataaaatta atttgatttt
    13141 tggaaataac caagcgacac cccatccgga gtcgacgggc ccatctggcc tgtgtttcag
    13201 acaccaggga gtcctaggag atctgcgaag atacggccac gggtgctctt gatcctgtgg
    13261 ctgattttgg actgtgctgc tcgcagctgc tgatgaatca catacttcct ccattttctt
    13321 ccactgattg actgttataa tttccctaat ttccaggtca aggtgctgtg cattgtggta
    13381 atagatgtga catgacgtca cttccaaagg accaatgaac atgtctgacc aatttcatat
    13441 aatgtgaaaa cgattttcat aggcagaata aataacattt aaattaaact gggcatcagc
    13501 gcaattcaat tggtttggta atagcaaggg aaaatagaat gaagtgatct ccaaaaaata
    13561 agtacttttt gactgtaaat aaaattgtaa ggagtaaaaa gtactttttt ttctaaaaaa
    13621 atgtaattaa gtaaaagtaa aagtattgat ttttaattgt actcaagtaa agtaaaaatc
    13681 cccaaaaata atacttaagt acagtaatca agtaaaatta ctcaagtact ttacacctct
    13741 ggttcttgac cccctacctt cagcaagccc agcagatcca ctagttctag agcggccgcc
    13801 accgcggtgg agctccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta
    13861 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
    13921 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
    13981 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
    14041 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
    14101 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
    14161 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
    14221 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
    14281 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
    14341 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
    14401 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
    14461 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
    14521 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
    14581 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
    14641 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
    14701 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
    14761 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
    14821 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
    14881 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
    14941 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
    15001 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
    15061 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
    15121 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
    15181 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
    15241 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
    15301 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
    15361 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
    15421 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
    15481 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
    15541 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
    15601 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
    15661 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
    15721 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
    15781 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
    15841 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
    15901 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
    15961 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tg
//
