LOCUS       sequence-429738-       20687 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For making transgenic fish.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 20687)
  AUTHORS   Bertrand JY, Chi NC, Santoso B, Teng S, Stainier DY, Traver D
  TITLE     Haematopoietic stem cells derive directly from aortic endothelium 
            during development.
  JOURNAL   Nature. 2010 Mar 4;464(7285):108-11. doi: 10.1038/nature08738. Epub 
            2010 Feb 14.
  PUBMED    20154733
REFERENCE   2  (bases 1 to 20687)
  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"; doi: 
            "10.1038/nature08738"; journalName: "Nature"; date: "2010-03-4- 4"; 
            volume: "464"; issue: "7285"; pages: "108-11"
COMMENT     Sequence Label: bactin2:loxP-STOP-loxP-DsRed-express
FEATURES             Location/Qualifiers
     source          1..20687
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     50..69
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        50..68
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    join(96..107,108..295)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     polyA_signal    734..868
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(879..1556)
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAEGRHH
                     LFL"
     primer_bind     complement(941..964)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(1139..1158)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1356..1376
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     1408..1426
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      1553..1562
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    1597..1630
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      1661..1670
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1695..1816
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(2241..2261)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          2391..2456
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     polyA_signal    2548..2669
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(3094..3114)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          3244..3309
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     polyA_signal    3401..3522
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(3947..3967)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          4097..4162
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     promoter        4772..5271
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(4807..4830)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     5204..5223
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             5315..5914
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5315..5334)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5811..5831
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     protein_bind    6455..6488
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     9254..9270
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     9255..9271
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    join(17670..17832,17833..17844)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     CDS             17856..17867
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(17877..17897)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(17877..17895)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(17939..17955)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(17939..17955)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(17952..17974)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    17963..17979
                     /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(17987..17993,17994..18011,18012..18017))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    18032..18053
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(18170..18187)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(18341..18929)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(18421..18440)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(19100..19891,19892..19960))
                     /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     19723..19742
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(19961..20065)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(20091..20546)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(20228..20249)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     20440..20459
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     join(20673..20687,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(20687,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgag cgcgtaggga taacagggta atgcgcgcgt aatacgactc
       61 actatagggc gaattgggta ccgggggtca agaaccagag gtgtaaagta cttgagtaat
      121 tttacttgat tactgtactt aagtattatt tttggggatt tttactttac ttgagtacaa
      181 ttaaaaatca atacttttac ttttacttaa ttacattttt ttagaaaaaa aagtactttt
      241 tactccttac aattttattt acagtcaaaa agtacttatt ttttggagat cacttcattc
      301 tattttccct tgctattacc aaaccaattg aattgcgctg atgcccagtt taatttaaat
      361 gttatttatt ctgcctatga aaatcgtttt cacattatat gaaattggtc agacatgttc
      421 attggtcctt tggaagtgac gtcatgtcac atctattacc acaatgcaca gcaccttgac
      481 ctggaaatta gggaaattat aacagtcaat cagtggaaga aaatggagga agtatgtgat
      541 tcatcagcag ctgcgagcag cacagtccaa aatcagccac aggatcaaga gcacccgtgg
      601 ccgtatcttc gcagatctga tctagaggat cataatcagc cataccacat ttgtagaggt
      661 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
      721 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
      781 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
      841 catcaatgta tcttatcatg tctggatcct cgaggccgct acaggaacag gtggtggcgg
      901 ccctcggcgc gctcgtactg ctccacgatg gtgtagtcct cgttgtggga ggtgatgtcc
      961 agcttggagt ccacgtagta gtagccgggc agctgcacgg gcttcttggc catgtagatg
     1021 gacttgaact ccaccaggta gtggccgccg tccttcagct tcagggcctt gtggatctcg
     1081 cccttcagca cgccgtcgcg ggggtacagg cgctcggtgg aggcctccca gcccatagtc
     1141 ttcttctgca ttacggggcc gtcggagggg aagttcacgc cgatgaactt caccttgtag
     1201 atgaaggagc cgtcctgcag ggaggagtcc tgggtcacgg tcaccacgcc gccgtcctcg
     1261 aagttcatca cgcgctccca cttgaagccc tcggggaagg acagcttctt gtagtcgggg
     1321 atgtcggcgg ggtgcttcac gtacaccttg gagccgtact ggaactgggg ggacaggatg
     1381 tcccaggcga agggcagggg gccgcccttg gtcaccttca gcttggcggt ctgggtgccc
     1441 tcgtaggggc ggccctcgcc ctcgccctcg atctcgaact cgtggccgtt cacggagccc
     1501 tccatgcgca ccttgaagcg catgaactcc ttgatgacgt cctcggagga ggccatggtg
     1561 gcgaccggtg gatcccgggc ccgcggtacc gtcgacataa cttcgtataa tgtatgctat
     1621 acgaagttat taggtccctc gacctgcaga cttactcaag ccatggtggc tagcttgcgg
     1681 aattccagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     1741 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     1801 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     1861 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg ctgattatga
     1921 tctctagtca aggcactata catcaaatat tccttattaa cccctttaca aattaaaaag
     1981 ctaaaggtac acaatttttg agcatagtta ttaatagcag acactctatg cctgtgtgga
     2041 gtaagaaaaa acagtatgtt atgattataa ctgttatgcc tacttataaa ggttacagaa
     2101 tatttttcca taattttctt gtatagcagt gcagcttttt cctttgtggt gtaaatagca
     2161 aagcaagcaa gagttctatt actaaacaca gcatgactca aaaaacttag caattctgaa
     2221 ggaaagtcct tggggtcttc tacctttctc ttcttttttg gaggagtaga atgttgagag
     2281 tcagcagtag cctcatcatc actagatggc atttcttctg agcaaaacag gttttcctca
     2341 ttaaaggcat tccaccactg ctcccattca tcagttccat aggttggaat ctaaaataca
     2401 caaacaatta gaatcagtag tttaacacat tatacactta aaaattttat atttacctta
     2461 gagctttaaa tctctgtagg tagtttgtcc aattatgtca caccacagaa gtaaggttcc
     2521 ttcacaaaga tccctcgacc agacatataa gatacattga tgagtttgga caaaccacaa
     2581 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     2641 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     2701 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     2761 tggctgatta tgatctctag tcaaggcact atacatcaaa tattccttat taaccccttt
     2821 acaaattaaa aagctaaagg tacacaattt ttgagcatag ttattaatag cagacactct
     2881 atgcctgtgt ggagtaagaa aaaacagtat gttatgatta taactgttat gcctacttat
     2941 aaaggttaca gaatattttt ccataatttt cttgtatagc agtgcagctt tttcctttgt
     3001 ggtgtaaata gcaaagcaag caagagttct attactaaac acagcatgac tcaaaaaact
     3061 tagcaattct gaaggaaagt ccttggggtc ttctaccttt ctcttctttt ttggaggagt
     3121 agaatgttga gagtcagcag tagcctcatc atcactagat ggcatttctt ctgagcaaaa
     3181 caggttttcc tcattaaagg cattccacca ctgctcccat tcatcagttc cataggttgg
     3241 aatctaaaat acacaaacaa ttagaatcag tagtttaaca cattatacac ttaaaaattt
     3301 tatatttacc ttagagcttt aaatctctgt aggtagtttg tccaattatg tcacaccaca
     3361 gaagtaaggt tccttcacaa agatccctcg accagacata taagatacat tgatgagttt
     3421 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     3481 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     3541 cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc
     3601 tacaaatgtg gtatggctga ttatgatctc tagtcaaggc actatacatc aaatattcct
     3661 tattaacccc tttacaaatt aaaaagctaa aggtacacaa tttttgagca tagttattaa
     3721 tagcagacac tctatgcctg tgtggagtaa gaaaaaacag tatgttatga ttataactgt
     3781 tatgcctact tataaaggtt acagaatatt tttccataat tttcttgtat agcagtgcag
     3841 ctttttcctt tgtggtgtaa atagcaaagc aagcaagagt tctattacta aacacagcat
     3901 gactcaaaaa acttagcaat tctgaaggaa agtccttggg gtcttctacc tttctcttct
     3961 tttttggagg agtagaatgt tgagagtcag cagtagcctc atcatcacta gatggcattt
     4021 cttctgagca aaacaggttt tcctcattaa aggcattcca ccactgctcc cattcatcag
     4081 ttccataggt tggaatctaa aatacacaaa caattagaat cagtagttta acacattata
     4141 cacttaaaaa ttttatattt accttagagc tttaaatctc tgtaggtagt ttgtccaatt
     4201 atgtcacacc acagaagtaa ggttccttca caaagatccc tcgatcgaga aaaaaaatat
     4261 aaaagagatg gaggaacggg aaaaagttag ttgtggtgat aggtggcaag tggtattccg
     4321 taagaacaac aagaaaagca tttcatatta tggctgaact gagcgaacaa gtgcaaaatt
     4381 taagcatcaa cgacaacaac gagaatggtt atgttcctcc tcacttaaga ggaaaaccaa
     4441 gaagtgccag aaataacagt agcaactaca ataacaacaa cggcggctac aacggtggcc
     4501 gtggcggtgg cagcttcttt agcaacaacc gtcgtggtgg ttacggcaac ggtggtttct
     4561 tcggtggaaa caacggtggc agcagatcta acggccgttc tggtggtaga tggatcgatg
     4621 gcaaacatgt cccagctcca agaaacgaaa aggccgagat cgccatattt ggtgtccccg
     4681 aggatccctg tggaaaaaaa agggacagga taagtatgac atcatcaagg aaaccctgga
     4741 ctactgcgcc cttcgacctc gaaattctac cgggtagggg aggcgctttt cccaaggcag
     4801 tctggagcat gcgctttagc agccccgctg ggcacttggc gctacacaag tggcctctgg
     4861 cctcgcacac attccacatc caccggtagg cgccaaccgg ctccgttctt tggtggcccc
     4921 ttcgcgccac cttctactcc tcccctagtc aggaagttcc cccccgcccc gcagctcgcg
     4981 tcgtgcagga cgtgacaaat ggaagtagca cgtctcacta gtctcgtgca gatggacagc
     5041 accgctgagc aatggaagcg ggtaggcctt tggggcagcg gccaatagca gctttgctcc
     5101 ttcgctttct gggctcagag gctgggaagg ggtgggtccg ggggcgggct caggggcggg
     5161 ctcaggggcg gggcgggcgc ccgaaggtcc tccggaggcc cggcattctg cacgcttcaa
     5221 aagcgcacgt ctgccgcgct gttctcctct tcctcatctc cgggcctttc gacctgcatc
     5281 catctagatc tcgatcgagc agctgaagct taccatgacc gagtacaagc ccacggtgcg
     5341 cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga
     5401 ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct
     5461 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
     5521 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
     5581 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
     5641 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
     5701 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
     5761 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc
     5821 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg
     5881 cacctggtgc atgacccgca agcccggtgc ctgacgcccg ccccacgacc cgcagcgccc
     5941 gaccgaaagg agcgcacgac cccatgcatc gatgatatca gatccccggg atgcagaaat
     6001 tgatgatcta ttaaacaata aagatgtcca ctaaaatgga agtttttcct gtcatacttt
     6061 gttaagaagg gtgagaacag agtacctaca ttttgaatgg aaggattgga gctacggggg
     6121 tgggggtggg gtgggattag ataaatgcct gctctttact gaaggctctt tactattgct
     6181 ttatgataat gtttcatagt tggatatcat aatttaaaca agcaaaacca aattaagggc
     6241 cagctcattc ctcccactca tgatctatag atctatagat ctctcgtggg atcattgttt
     6301 ttctcttgat tcccactttg tggttctaag tactgtggtt tccaaatgtg tcagtttcat
     6361 agcctgaaga acgagatcag cagcctctgt tccacataca cttcattctc agtattgttt
     6421 tgccaagttc taattccatc agaagctggt cgagataact tcgtataatg tatgctatac
     6481 gaagttatgt cgactgcaga attcgaagct tgagctcgag atccggctga actgtaaaag
     6541 aaagggaaac tgttagaatt gcactgctgc cagggagagg cgtttcagca agcatgtgac
     6601 catctggagt caacttcctg ttttgctctg cattctacag tctgaccgct ggtgggcgct
     6661 agtgacagcg cgcgcctcag atatgccaga caaaggaagt ccctctgcat tctctcatta
     6721 ttaccataaa aggcaatggt ttgagccgct ttgcggctgc cggcggtgtg gcactcagcc
     6781 agagtcgtag ggctcagtaa aaaggaacag atttcgacat tcatcacacc aggaccactc
     6841 cctaaatttt acacaaatga agtttcacat cctcttccac cctcgcctgc gatcaattac
     6901 acaaccgcat ttcaacgggt gtctgccgcc gggtgttact gcgtcacaga tctgcagtca
     6961 gtccacggac tcctcgtgag cggagacttc acactaaagc atttctggac ggtcactctc
     7021 agtcggttca gttcatagtc tagcctcaaa aatcaccgag atcctgaaaa actgatgggc
     7081 tctgcttcga cagtcacagc ggctgaagtg cagccagaga actccgcagc cgttattaaa
     7141 gtggaccaca cctctaaaat tcaaatcttt catagcggat acaacacttc actaacgtta
     7201 acgttaatct tcagcttcat taaagacaga gggggctgca ggctacattt aaatagaaca
     7261 cataacaaaa agaaaattaa ccaagttaaa actaaatgcc aatacccgtt actaataaca
     7321 agatcttcga tcttataaag cacgaatgag ccgatgttag attgatgggg tttacgttac
     7381 atcagatgag agctggttag ccggctagct aaagtaggtc acaccctgct gcagcactgc
     7441 ctgcacaaag aacttcccat gccggctatc catcaagtgt gcaacatgct caccagcacc
     7501 ctcaaaactg tagttaaaaa tcacatttat ggctgttaaa gagagattag ccagaacaca
     7561 actttatatt aagacttaac acccatttca caccggccta ttcaatgttc aatgtcgtgc
     7621 ctacgactgg tcacgttaaa ttaccgacat tctcacctgc cacatccact aacgctacaa
     7681 aaaagttaag atacagccaa tatttattaa atgtaacata gattatcaaa agaaccacca
     7741 cttataaatc aaacctagta gataactaca aagagaccgt tatgaactgt ctaaagactt
     7801 catcaaatca tatttatcca tttacacatt ttacttaaca ttaatagata gaaactataa
     7861 ttaaacgctt aaaaactgta aaaccaactc accttgggtt tggggggttg tcgacttaag
     7921 agaatgggtt acaagcagat gatattccgc actagctgcg tgtggaggag ctcaaagtga
     7981 ggctgaaaag ctgggcgcgc tcttttatat gacggcacat acccggctct agccataaaa
     8041 ggtaaacttt cggagctctg tgctctgatt ggtccagcgt cacacacccg gctgtcgcgt
     8101 gcacattgct acacttgctt cttgcaactt gagagtgaaa gaaggggaag cttaactggt
     8161 taaagaacaa ggccatggcg atttcctata aaactggaat ttttatttgc aaattttaac
     8221 acagaatttt tcgtattttt ctgcgacaac gccaagactc aattaaactt ttcaaaaagt
     8281 ctgtgcagag ttacaagtgg cagaatgcgt cataatgatg gatgcctcca tgatcctgtg
     8341 ttgtgttctc aatctaacac aacagcagct ctgtaggtca tttacctctt gtgttcacga
     8401 aataccgtaa tacaaacaat gggggatcta atgagtactt ttccatgtat aaaatggaca
     8461 ttttttttat ccaaattagg atgacactac ttgtagtatg ttgtatgagc aactagctgg
     8521 tcttagctca gtagtaatgc actttaagaa agtgctttaa taactttgaa tgaatgggac
     8581 ctgcattcca tgccacattg cttcactatt tttgtgtgta caagttacac aatgatgtat
     8641 aaagtgagtg atagcagtta catttttgca agtgggccta cctcaggttt aatgttcatt
     8701 agaaatgtta tatgtcaagt cttgatctta aatagttaaa ttttctgttc ataatttcat
     8761 gtcgtgtcta tgcagacata acatttcttt ttaaagaggt ttataaggat aaatcctaag
     8821 aacataattc tcattaaaag tagaaaaaaa aaacactgaa agtgttggtg atcagctaca
     8881 ataaataata ttttgcacac atttcagata ttgccacact gaataacaga taaagctgtg
     8941 cttgagcagc agtgatcatg ataaactaca tgtgactttt tactgtgtaa cttgggagta
     9001 atgcacatgg ccttcagtgg gagttataaa caaaaacagc tgattgtgtt aaaagtgctt
     9061 gctgatacca aggacaatga ggtgatgtga cttttatttt gtttttggca aattccaagt
     9121 attttgacaa gactttttac aattctgagc attttacctc taaaatatgg aagtttctca
     9181 gagtaaacgt tgagtatagg taatgactag gcccacacag aatctgtgcg cacagaaatc
     9241 tgcagatgcg gcctcgaggt cgacggtatc gataagcttg atatctgctg ataccaagga
     9301 caatgaggtg atgtgacttt tattttgttt ttggcaaatt ccaagtattt tgacaagact
     9361 ttttacaatt ctgagcattt tacctctaaa atatggaagt ttctcagagt aaacgttgag
     9421 tataggtaat gactaggccc acacagaatc tgtgcgcaca gaaatctgca gatttcccca
     9481 gatctttagc ccatcattga gtttatttat tgacttgtgt atttggattt gctctttagt
     9541 gtttggactc tcagcagtaa aaattaactc acactgaact aaactgaact gaacttaaac
     9601 tcttaaaaaa tggactgaca cagtttcaat ttactatact aatcctctaa gttaagctgc
     9661 tttgacacaa tctacattgt aaaagcactc tagaaataaa gacaaaggtg taaatatatg
     9721 tttattcagt ttttaaatac atttccgtaa tattattgac taatatgaaa gtgttcattt
     9781 gattacaata tggtctgaaa gtaatgtttt ctgtctttta gaagatatat tatgtgagag
     9841 acttgctttg tcgaccaaat aattggatct aggtggactt gcattgtata tatttattaa
     9901 aagttaaaaa ggtattattt ttttttattt catatagtaa gtttttagtt acgatactcc
     9961 cgaaaaaatc tcgcataaat cacagattgt ttacaaaatt ctctgcagaa aaagcaaaaa
    10021 atgtcagcag attctgtctg gccctagcta tgacccaaca gagcaaacag aataaataca
    10081 acgaaattta atatacagca tcagatataa aagtgaatgt tttatagctg tttgtgtata
    10141 gaaaaaatga gaaaaaacag acataaacag catcatagta tgaatgtcta aaaacttgtt
    10201 atctgcaaac tggatactca tcaataaggt ctcctccttc catcccccaa catcttttca
    10261 aagaatgcca agcatccatc cacacccccc tttataaaca cacaggtttt tataagataa
    10321 gcaaatattt agcaccaatc tgtggttaat gtgatgtaac acacaacaaa caatgcattc
    10381 agcatatcca gtcaattcta ttttttaacc aaccatgctt ctctttgtaa cttttgtaga
    10441 ttgcacaggg gatcttgtta cagcagatgt gcttttgtga gtccatagtt cccccgagga
    10501 ccaccaccct tccctgatag cagcccacta cagccttccc attcatctct aaataaggta
    10561 ctgtcctaat gaacctcagc tttgccccat gacataagat ttccagagca ataaagtgtt
    10621 gttgactgtc agtgtgataa tgtcattgtc ctttagaatg ccaaagtctc ttatctcaat
    10681 tctgatacca aagctctccc agtacaccat tcatgcataa tgataagaaa catatttatg
    10741 aagttgttct caaaggaaca ctttttgggc gtgtcatgct gcatttcata gtttgttgtt
    10801 tggcacccac ataaataaaa caagtgtgtt tttgataata ataataataa gggatgcacc
    10861 gataccagta tcaggttttg ggtcgatact tggttaaaat attcttactc gtattgcaca
    10921 tgaaatgctg aaaccacgca ccaataccac tcaacaagat ttcattacat ggatgtgatt
    10981 tgttgtccta cctgacatcg atgtcttgat gtattaatga atgtctgata tattaatagc
    11041 ttagtcttta taaaccatcc gtttataaaa tacactttta acaaatctat aggcttctga
    11101 cgaataaacc gaatattcaa tatgatcttt gatcatgtaa tgtgtaaagt tgtgatggcg
    11161 caagcataac ttatccttta aatgtccacg tttaatcggt aatcgataca gggttgtcaa
    11221 caacgtgcgt atattcttag cttaaaatat ccttctggta aagatttttg tgttcatcct
    11281 tgcgacaggc aagaaataaa accttctttc aaattaatta cattgaaacc aaatccaaga
    11341 agtatcagaa tcggtactca gtattgtcga gtaacagaaa ctattatact tgtactcata
    11401 tcggtttgta aaaaagtggt atcggtttat ctctaatcat tcattcattc attcattttc
    11461 aacttagtcc tttaattaac caggggtcac cataggggaa tgaaccgcca acttatcctg
    11521 catatgtttt acacagcaga tgtccttcca gttgcaacct aacactggga aacacctata
    11581 cactctcatt cacacacaca cacacactca ctatggtcaa tttagcttac cccattcacc
    11641 tatacatcat gtctttggac ttgtgagggg aaaccagagc accgggagga aacctacttg
    11701 aacagggaga acatgcaaac tccacacaga actgccaact gatccagccg gggctctaaa
    11761 cagcaaccta cttgctgtga ggtaattgtg ccacccatcg cgccaccgtg atgtccatcc
    11821 ctaataataa taatacagtt tatttatagt gcgcttttct cagacccaaa gcgctacaat
    11881 gcatacaagg caaacagtaa caaagcagaa caataaaaaa gctgtaaaaa caccacaata
    11941 agacaataaa atatgaacaa tacaatgtaa ttattaaatt gacaaaaata atcaacctaa
    12001 attaagacca acagtgaaaa ggtgagtctt gaaaagtttc ttcgaactgg aattcctgat
    12061 attttttctt tccttatttt cttttatgta tttttttttt accctccttg tcttgggaat
    12121 agcggaagcg aagcccttca tttgtcctat aaacagcaga aggaccaccg attgatttta
    12181 gattgacttg ttcttggttg cttgatggtg cagatactca ggcttgaaag gaagtcacat
    12241 atatactgac agctggcctc ttagtctaga gattggatta accccctaca ttatgaccca
    12301 aatggattca tatgtggtta ctcatcgcct tatatgagtg taaataatta gatacttttc
    12361 tcttatgcaa tattatgtga caaatcatta tttaagatac atcatagtgg gtttacattc
    12421 aaaatggtgt tacttcagca gacaaagaca tattttgtgt cttcatttgt attgtcaaac
    12481 cacttgcaac aggatttgat actaatcaag aaaaaaaagc cttgttcctc atttatgttc
    12541 ctcagcagaa cagttgtcac cgtgtccttt cttgccattg caaagcttca cttcctcttt
    12601 gctgtaaaag ctggtagggg aggaccgcta gtccatttat ggctgtttgc ttgcatgtac
    12661 aaaattgtca acatattgct ttgcatactt gggtttgtca cttgtcatat tttagcggta
    12721 gttcctgcga agattaacgt tttgttatta cattgaatga ttaggttgca gctgggaggg
    12781 catccatatg ctggataagt tggcggttca ttccgctgtg gctacttttg atgaataaag
    12841 ggattaagcc aaagaaaatg aatgaatgat tgaatgatta tttctaaaca cttattctac
    12901 tgacaaaaac agtcaaaaaa tatttttgtt ttgtgtttca cagaagacag tcaccaaggt
    12961 ttggagtgac aaagtggaaa acttgtacat tttggggtga actatctctt taaggctgca
    13021 tatgctttca aaatgtgtac ataatgctct ctgtataaga aagttctgct cgcttcagtg
    13081 ctggctgatc aatagagatc aagggcttat tagcacccca ctgtgcccac tgctaaacag
    13141 gggctcaatc tctgctactg agatataaat aagtcatgtt gggatcattt caacatgctg
    13201 ctctagagat tgcatcaatc atattcataa tttgtcaact ccaagcgcca tggttcataa
    13261 tttgtcaact ccaagcgtta cgattgcata gttttgaaca tttgacactg attataacat
    13321 ttaatgcatg ctacacattg aatccaccac tatgtggcct gcagttatct cctccaaatg
    13381 catggggatt tacccagagc tgaagggggt tatgatttgg gaccactgtg tttgggtcag
    13441 tttctagcaa tatctgcttt tgtaccaaga ccctaggcct cctaatctag cctccaccat
    13501 ctgtctgccc gtgagagaga gagaaggctg agcaggggat tggccctgga atgagtcaga
    13561 cgtgctcgtc cacacttcag actcagtcag cctttaaacc agcatgggag agattactgt
    13621 gccaagcacc atagcttggg cctgccacag gaagctgagt aattcaagca gatactttag
    13681 cataatagat ttttatgtgt tcaagagcag ttttaacatt caaaatatgt agtgcaagac
    13741 ggaaatgatc ctcttttaaa ggcatagttt acccaacaat aaacactgtc attatttact
    13801 caccgtggtg ttgtctttag ccttatttta aatgaccgtt aattgggtaa gttaaattgt
    13861 ccgtagtgtg tgtgtgaatg agagtgtatg gatgttttcg agggatgggt tgcggctgga
    13921 agagcatctg ctgcataaaa catgctagat aagttggtgg ttcattccgc tgtggcaact
    13981 cctgataaat aaacggacta agctgtaaat aaaatgaaca aatgaccgtt aaataaatga
    14041 attaggggct ttttcatttc tgaaagacaa atatgcaatc tttaaggggg atatagatat
    14101 tgccttcaag aatgtgtgct tatagtcact ttaaataatc atgttagaca tttgcaaaac
    14161 aaattggctc caccgaaacc gtaatgtaaa taacaaaaga ttgccgccat tcattttgtt
    14221 tgccgttaat accttattca ttagagcttt gttgctcaag agtttcatgt taatgtctct
    14281 tgcgcatgaa atatctgtca ttcctgagtg atgtctgtgt gaaactgatg aggtgtggcg
    14341 ctcacagcat ctctggctag actgaggtat cccaaggccc aggtccaaca ttaatattgg
    14401 cagaggttca tattagggtc atttgtgttg aagtgaatac atcaactttc agcccacctt
    14461 taatagcact gccagtcttg ctgaatgttt ggcattgatc taataattaa gtctttttaa
    14521 gcttggagtt tcaacatatt cgatctggta aagtaaatat gttgattgaa ataaagctct
    14581 ttccttaaac gatgtaaatt ccactccctg aaaaccaaag gccggtgtta gttgccaaat
    14641 aagggaatga cctatgcatc tcttacagga agtgcgtttt taaaggatgt catttcttca
    14701 gcaagtttgc acaggaaatc gcattgaaat attctgctgt gggagaaagc ttagccaact
    14761 aaggaatgta caacttagta gtagtgctta ctccttttat cgtgatgctg tgcagtgcac
    14821 acacacctca ctctcactta tgtgtggaaa gaaacaacag tacagcatgc aaacaagatt
    14881 ttaaatgact tcgtaaacag ctgtttgcct catctgggcg cttaattagc tccagcagaa
    14941 tctagcatag agtttataaa aactttcagt ttaattatgt tgtctgcttt ctataattaa
    15001 tgtaattctt acattaaagg gctttttatt tgttcagttg ttttatgaat agttaatcct
    15061 taatataatg ctgtgctaaa aaacaaataa ctgcttaatt ttgggctcaa tatagcattt
    15121 gtgttttgtt ttttgttatg agtaatctaa aggaaaaccc tgctgaggtt ttctgatgac
    15181 tataactctt tgccacaatc cttacaagtc atttgccctt tctataatga tgcattggct
    15241 atactaagag gcaacagaca tgttctttct tgttgagcaa ttcagaagct ttttggctta
    15301 ggtgtggcac aaaactctgg aaaatgactt gcacataatg tgcatatgat ttagagaaat
    15361 ataacatctt aaatataaaa aatatatgaa aagcattgta aaattgtcaa tattataata
    15421 tacctccagt actataaaac tccatgacat gtaagtatga tagcaacgct tgacatcttt
    15481 agaaatgtga agaatttcga tgcaaagata tgagatacgg cataacaaga catcctgcta
    15541 ggcatgcgct gcatttttcc tcaaactttg tccacaaaag gattcttcat atttatttcc
    15601 tcaagggctt tagctgtaga ccgttctgtg cactttggtt gtgttaggcc tcgccctctg
    15661 gtcgatgtaa cagcaaccgc cataaaagga aatctttaag tgtgtcacat ggtaacaatg
    15721 tcaagaattc aaggcccttg tcacatggga ctttctcatg gaaatcagat tgaaacttct
    15781 aggcctaacg aatgaagaaa cgctaagtta gagaagctgg ggagttttga tcaaacaatg
    15841 acatttattt tatttctggc ttgctgtagt ttcactctgc acactgtgag actctttcat
    15901 tgacacatat tcgtatgtaa aatccatata tattcagctt ttacatcata cagtaatgtt
    15961 ccttgtttgt ttgtagcatt ttcatttgtt gtgtctttct gccaaagcaa cagtgtttct
    16021 cattcattcc tccatttgaa gggttgccta tgatctgcct cacgatgagt aagaaggaag
    16081 tggtttgaaa taatgacagc gaggtcatat ccatatatag tcagcctttg gcaaggcaat
    16141 gatgactggt gatgtctcaa cttcagaaag ctgatttcat atcactgttt ttcctcaatg
    16201 tcctgttcac attgatttgc tcgctttgca ctttcacaca ctaacgcttg acacagtcaa
    16261 taagcccaga taaactatca acagtctgtc gttccctttc agaaactctc tcactcttaa
    16321 tctgcctctc ccaaacacgc acacgttctt actcactcac accatcactc tcatgctccg
    16381 cccgatctaa cactgtttag aaaccaaaca attaccagta tgcgagtgga tgggcggagc
    16441 ttagctttat ctccagcttc tcaggctcca aataaggaaa ggactagcag gtagcgattg
    16501 ccaaacttag tcatcaaagc aagcatctca ttcattgcac actcagcttg tgttttgctt
    16561 atgagaagct ctgagatctg tctgctttgc aaaattccat caaataacat gctaatggat
    16621 ttattagcga ttgaggtctg tgaatgagtt tggagttgct tgtcacttcc ttcgactgga
    16681 tttgcaagag gaaatggcat tgtgttgtga cagaagtcat gtatcttagg aactgacttg
    16741 tggatgtggc cacagcaggt tttcctttgc ttttcagggc ggtaaattcc tttctgcaga
    16801 ttgttatatc gctgttgatc atttactgaa ccaatgtcat gttgaagatt gaagtacact
    16861 aaagagttcc tctgttgaga agaaaacaac actcagtgac caaatttgtc agtaaatagt
    16921 tgtgaatgat tatgtcagtg cctttaccct cataggcatc tgatacatgg caatttatgc
    16981 aaatggcaca tgagatacaa ggtcatattt ggatttcaca tagagtcaag gatgaaatta
    17041 ttcatacccc tggccactgt tggggaaagt tactttagaa agttatgtat tacattattt
    17101 agttaaaaaa gtaactgagc tacttatatt taatatttat ggtaagtaat gtgttaaatt
    17161 acttttgagt tctttctgga ttattttctc tgatctggct gaagtttgat ctcttttaaa
    17221 acctaatgac aatacttcta aataatgaga atacgaagaa atgtaaggct ctaccctgtc
    17281 tgttcttgcg cggccgccac cgcggcccat ctggcctgtg tttcagacac cagggagtct
    17341 ctgctcacgt ttcctgctat ttgcagcctc tctatcaaga ctaatacacc tcttcccgca
    17401 tcggctgcct gtgagaggct tttcagcact gcaggattgc ttttcagccc caaaagagct
    17461 aggcttgaca ctaacaattt tgagaatcag cttctactga agttaaatct gaggttttac
    17521 aactttgagt agcgtgtact ggcattagat tgtctgtctt atagtttgat aattaaatac
    17581 aaacagttct aaagcaggat aaaaccttgt atgcatttca tttaatgttt tttgagatta
    17641 aaagcttaaa caagaatctc tagttttctt tcttgctttt acttttactt ccttaatact
    17701 caagtacaat tttaatggag tactttttta cttttactca agtaagattc tagccagata
    17761 cttttacttt taattgagta aaattttccc taagtacttg tactttcact tgagtaaaat
    17821 ttttgagtac tttttacacc tctgtcaaga actcctggga gctccagctt ttgttccctt
    17881 tagtgagggt taattgcgcg cattaccctg ttatccctac gcgcttggcg taatcatggt
    17941 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
    18001 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
    18061 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
    18121 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
    18181 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
    18241 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
    18301 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
    18361 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
    18421 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
    18481 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
    18541 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
    18601 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
    18661 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
    18721 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
    18781 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
    18841 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
    18901 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
    18961 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
    19021 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
    19081 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
    19141 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
    19201 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
    19261 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
    19321 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
    19381 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
    19441 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
    19501 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
    19561 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
    19621 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
    19681 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
    19741 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
    19801 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
    19861 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
    19921 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
    19981 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
    20041 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct aaattgtaag
    20101 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
    20161 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
    20221 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
    20281 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
    20341 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
    20401 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
    20461 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
    20521 gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca actgttggga
    20581 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
    20641 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgtt
//
