LOCUS       sequence-47281-e       17461 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 17461)
  AUTHORS   Zou J, Mali P, Huang X, Dowey SN, Cheng L
  TITLE     Site-specific gene correction of a point mutation in human iPS cells
            derived from an adult patient with sickle cell disease.
  JOURNAL   Blood. 2011 Oct 27;118(17):4599-608. Epub 2011 Aug 31.
  PUBMED    21881051
REFERENCE   2  (bases 1 to 17461)
  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: "Blood."; 
            date: "2011-10-27"; volume: "118(17)"; pages: "4599-608. Epub 2011 
            Aug 31"
COMMENT     Sequence Label: BD2
FEATURES             Location/Qualifiers
     source          1..17461
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1..17
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    complement(6001..6034)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        6045..6544
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(6080..6103)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     6477..6496
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             6562..7587
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             6562..7587
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    7897..8121
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    complement(8128..8161)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    9463..9857
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     promoter        10215..11393
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          10446..11384
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     11341..11361
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     promoter        11410..11428
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      12651..12660
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(12657..12659,12660..12662,12663..13376)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(12657..12659,12660..12662,12663..13373)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(12702..12723)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(12963..12982)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    13416..13640
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(13691..13709)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(13716..13733)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(13716..13732)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13725..13747)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      13873..14301
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(13960..13979)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     14170..14191
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             14645..15439
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(14699..14718)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     15309..15328
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(15457..15525,15526..16317)
                     /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(15675..15694)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      16488..17076
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     16977..16996
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     17230..17247
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    17364..17385
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(17400..17405,17406..17423,17424..17430)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    17438..17454
                     /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)."
     primer_bind     join(17443..17461,1..4)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 caggaaacag ctatgaccat gattacgcca agcttggtac cgagctcgga tccactagta
       61 acggccgcca gtgtgctgga attcgccctt atcggtaccg tgtgtaagaa ggttcctgag
      121 gctctacaga tagggagcac ttgtttattt tacaaagagt acatgggaaa agagaaaagc
      181 aagggaaccg tacaaggcat taatgggtga cacttctacc tccaaagagc agaaattatc
      241 aagaactctt gatacaaaga taatactggc actgcagagg ttctagggaa gacctcaacc
      301 ctaagacata gcctcaaggg taatgctacg attaaactcc aacaattact gagaaaataa
      361 tgtgctcaat taaaggcata atgattactc aagacaatgt tatgttgtct ttcttcctcc
      421 ttcctttgcc tgcacattgt agcccataat actatacccc atcaagtgtt cctgctccaa
      481 gaaatagctt cctcctctta cttgccccag aacatctctg taaagaattt cctcttatct
      541 tcccatattt cagtcaagat tcattgctca cgtattactt gtgacctctc ttgaccccag
      601 ccacaataaa cttctctata ctacccaaaa aatctttcca aaccctcccc gacaccatat
      661 ttttatattt ttcttattta tttcatgcac acacacacac tccgtgcttt ataagcaatt
      721 ctgcctattc tctaccttct tacatgccta ctgtgcctca tattaaattc atcaatgggc
      781 agaaagaaaa tatttattca agaaaacagt gaatgaatga acgaatgagt aaatgagtaa
      841 atgaaggaat gattattcct tgctttagaa cttctggaat tagaggacaa tattaataat
      901 accatcgcac agtgtttctt tgttgttaat gctacaacat acaaagagga agcatgcagt
      961 aaacaaccga acagttattt cctttctgat cataggagta atattttttt ccttgagcac
     1021 catttttgcc ataggtaaaa ttagaaggat ttttagaact ttctcagttg tatacatttt
     1081 taaaaatctg tattatatgc atgttgatta attttaaact tacttgaata cctaaacaga
     1141 atctgttgtt tccttgtgtt tgaaagtgct ttcacagtaa ctctgtctgt actgccagaa
     1201 tatactgaca atgtgttata gttaactgtt ttgatcacaa cattttgaat tgactggcag
     1261 cagaagctct tttatatcca tgtgttttcc ttaagtcatt atacatagta ggcactgaga
     1321 actcttcata tctgaataag atatttagga accactggtt tacatatcag aagcagagct
     1381 actcagggca ttttggggaa gatcactttc acattcctga gcatagggaa gttctcataa
     1441 gagtaagata ttaaaaggag atacttgtgt ggtattcgaa agacagtaag agagattgta
     1501 gaccttatga tcttgatagg gaaaacaaac tacattcctt tctccaaaag tcaaaaaaaa
     1561 agagcaaata tagcttacta taccttctat tcctacacca ttagaagtag tcagtgagtc
     1621 taggcaagat gttggcccta aaaatccaaa taccagagaa ttcatgagaa catcacctgg
     1681 atgggacatg tgccgagcac acacaattac tatatgctag gcattgctat cttcatattg
     1741 aagatgagga ggtcaagaga tgaaaaaaga cttggcacct tgttgttata ttaaaattat
     1801 ttgttagagt agagcttttg taagagtcta ggagtgtggg agctaaatga tgatacacat
     1861 ggacacaaag aatagatcaa cagacaccca ggcctacttg agggttgagg gtgggaagag
     1921 ggagacgatg aaaaagaacc tattgggtat taagttcatc actgagtgat gaaataatct
     1981 gtacatcaag acccagtgat atgcaattta cctatataac ttgtacatgt acccccaaat
     2041 ttaaaatgaa agttaaaaca aagtatagga atggaattaa ttcctcaaga tttggcttta
     2101 attttatttg ataatttatc aaatggttgt ttttcttttc tcactatggc gttgctttat
     2161 aaactatgtt cagtatgtct gaatgaaagg gtgtgtgtgt gtgtgaaaga gagggagaga
     2221 ggaagggaag agaggacgta ataatgtgaa tttgagttca tgaaaatttt tcaataaaat
     2281 aatttaatgt caggagaatt aagcctaata gtctcctaaa tcatccatct cttgagcttc
     2341 agagcagtcc tctgaattaa tgcctacatg tttgtaaagg gtgttcagac tgaagccaag
     2401 attctacctc taaagagatg caatctcaaa tttatctgaa gactgtacct ctgctctcca
     2461 taaattgaca ccatggccca cttaatgagg ttaaaaaaaa gctaattctg aatgaaaatc
     2521 tgagcccagt ggaggaaata ttaatgaaca aggtgcagac tgaaatataa atttttctgt
     2581 aataattatg catatacttt agcaaagttc tgtctatgtt gactttattg ctttttggta
     2641 agaaatacaa ctttttaaag tgaactaaac tatcctattt ccaaactatt ttgtgtgtgt
     2701 gcggtttgtt tctatgggtt ctggttttct tggagcattt ttatttcatt ttaattaatt
     2761 aattctgaga gctgctgagt tgtgtttact gagagattgt gtatctgcga gagaagtctg
     2821 tagcaagtag ctagactgtg cttgacctag gaacatatac agtagattgc taaaatgtct
     2881 cacttgggga attttagact aaacagtaga gcatgtataa aaatactcta gtcaagtgct
     2941 gcttttgaaa caaatgataa aaccacactc ccatagatga gtgtcatgat tttcatggag
     3001 gaagttaata ttcatcctct aagtataccc agactagggc cattctgata taaaacatta
     3061 ggacttaaga aagattaata gactggagta aaggaaatgg acctctgtct ctctcgctgt
     3121 ctcttttttg aggacttgtg tgtgtgtgtg tgtgtgtgtg tgtgtgttgt ggtcagtggg
     3181 gctggaataa aagtagaata gacctgcacc tgctgtggca tccattcaca gagtagaagc
     3241 aagctcacaa tagtgaagat gtcagtaagc ttgaatagtt tttcaggaac tttgaatgct
     3301 gatttagatt tgaaactgag gctctgacca taaccaaatt tgcactattt attgcttctt
     3361 gaaacttatt tgcctggtat gcctgggctt ttgatggtct tagtatagct tgcagccttg
     3421 tccctgcagg gtattatggg taatagaaag aaaagtctgc gttacactct agtcacacta
     3481 agtaactacc attggaaaag caacccctgc cttgaagcca ggatgatggt atctgcagca
     3541 gttgccaaca caagagaagg atccatagtt catcatttaa aaaagaaaac aaaatagaaa
     3601 aaggaaaact atttctgagc ataagaagtt gtagggtaag tctttaagaa ggtgacaatt
     3661 tctgccaatc aggatttcaa agctcttgct ttgacaattt tggtctttca gaatactata
     3721 aatataacct atattataat ttcataaagt ctgtgcattt tctttgaccc aggatatttg
     3781 caaaagacat attcaaactt ccgcagaaca ctttatttca catatacatg cctcttatat
     3841 cagggatgtg aaacagggtc ttgaaaactg tctaaatcta aaacaatgct aatgcaggtt
     3901 taaatttaat aaaataaaat ccaaaatcta acagccaagt caaatctgta tgttttaaca
     3961 tttaaaatat tttaaagacg tcttttccca ggattcaaca tgtgaaatct tttctcaggg
     4021 atacacgtgt gcctagatcc tcattgcttt agttttttac agaggaatga atataaaaag
     4081 aaaatactta aattttatcc ctcttacctc tataatcata cataggcata attttttaac
     4141 ctaggctcca gatagccata gaagaaccaa acactttctg cgtgtgtgag aataatcaga
     4201 gtgagatttt ttcacaagta cctgatgagg gttgagacag gtagaaaaag tgagagatct
     4261 ctatttattt agcaataata gagaaagcat ttaagagaat aaagcaatgg aaataagaaa
     4321 tttgtaaatt tccttctgat aactagaaat agaggatcca gtttcttttg gttaacctaa
     4381 attttatttc attttattgt tttattttat tttattttat tttattttgt gtaatcgtag
     4441 tttcagagtg ttagagctga aaggaagaag taggagaaac atgcaaagta aaagtataac
     4501 actttcctta ctaaaccgac atgggtttcc aggtaggggc aggattcagg atgactgaca
     4561 gggcccttag ggaacactga gaccctacgc tgacctcata aatgcttgct acctttgctg
     4621 ttttaattac atcttttaat agcaggaagc agaactctgc acttcaaaag tttttcctca
     4681 cctgaggagt taatttagta caaggggaaa aagtacaggg ggatgggaga aaggcgatca
     4741 cgttgggaag ctatagagaa agaagagtaa attttagtaa aggaggttta aacaaacaaa
     4801 atataaagag aaataggaac ttgaatgaag gaaatgattt taaaacgcag tattcttagt
     4861 ggactagagg aaaaaaataa tctgagccaa gtagaagacc ttttcccctc ctacccctac
     4921 tttctaagtc acagaggctt tttgttcccc cagacactct tgcagattag tccaggcaga
     4981 aacagttaga tgtccccagt taacctccta tttgacacca ctgattaccc cattgatagt
     5041 cacactttgg gttgtaagtg actttttatt tatttgtatt tttgactgca ttaagaggtc
     5101 tctagttttt tatctcttgt ttcccaaaac ctaataagta actaatgcac agagcacatt
     5161 gatttgtatt tattctattt ttagacataa tttattagca tgcatgagca aattaagaaa
     5221 aacaacaaca aatgaatgca tatatatgta tatgtatgtg tgtacatata cacatatata
     5281 tatatttttt ttttcttttc ttaccagaag gttttaatcc aaataaggag aagatatgct
     5341 tagaactgag gtagagtttt catccattct gtcctgtaag tattttgcat attctggaga
     5401 cgcaggaaga gatccatcta catatcccaa agctgaatta tggtagacaa aactcttcca
     5461 cttttagtgc atcaatttct tatttgtgta ataagaaaat tgggaaaacg atcttcaata
     5521 tgcttaccaa gctgtgattc caaatattac gtaaatacac ttgcaaagga ggatgttttt
     5581 agtagcaatt tgtactgatg gtatggggcc aagagatata tcttagaggg agggctgagg
     5641 gtttgaagtc caactcctaa gccagtgcca gaagagccaa ggacaggtac ggctgtcatc
     5701 acttagacct caccctgtgg agccacaccc tagggttggc caatctactc ccaggagcag
     5761 ggagggcagg agccagggct gggcataaaa gtcagggcag agccatctat tgcttacatt
     5821 tgcttctgac acaactgtgt tcactagcaa cctcaaacag acaccatggt gcacctgact
     5881 cctgaggaga agtctgccgt tactgccctg tggggcaagg tgaacgtgga tgaagttggt
     5941 ggtgaggccc tgggcaggtt ggtatcaagg ttacaagaca ggtttaagga gaccgctagc
     6001 ataacttcgt atagcataca ttatacgaag ttatgaattc taccgggtag gggaggcgct
     6061 tttcccaagg cagtctggag catgcgcttt agcagccccg ctgggcactt ggcgctacac
     6121 aagtggcctc tggcctcgca cacattccac atccaccggt aggcgccaac cggctccgtt
     6181 ctttggtggc cccttcgcgc caccttctac tcctccccta gtcaggaagt tcccccccgc
     6241 cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca ctagtctcgt
     6301 gcagatggac agcaccgctg agcaatggaa gcgggtaggc ctttggggca gcggccaata
     6361 gcagctttgc tccttcgctt tctgggctca gaggctggga aggggtgggt ccgggggcgg
     6421 gctcaggggc gggctcaggg gcggggcggg cgcccgaagg tcctccggag gcccggcatt
     6481 ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc tcttcctcat ctccgggcct
     6541 ttcgacctgc atcccgccac catgaaaaag cctgaactca ccgcgacgtc tgtcgagaag
     6601 tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctcgga gggcgaagaa
     6661 tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc
     6721 gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg
     6781 attccggaag tgcttgacat tggggaattc agcgagagcc tgacctattg catctcccgc
     6841 cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc tgttctgcag
     6901 ccggtcgcgg aggccatgga tgcgatcgct gcggccgatc ttagccagac gagcgggttc
     6961 ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt catatgcgcg
     7021 attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc
     7081 gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga agtccggcac
     7141 ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg cataacagcg
     7201 gtcattgact ggagcgaggc gatgttcggg gattcccaat acgaggtcgc caacatcttc
     7261 ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga gcggaggcat
     7321 ccggagcttg caggatcgcc gcggctccgg gcgtatatgc tccgcattgg tcttgaccaa
     7381 ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca gggtcgatgc
     7441 gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc ccgcagaagc
     7501 gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc
     7561 agcactcgtc cgagggcaaa ggaatagagt agatgccgac cgaacaagag ctgatttcga
     7621 gaacgcctca gccagcaact cgcgcgagcc tagcaaggca aatgcgagag aacggcctta
     7681 cgcttggtgg cacagttctc gtccacagtt cgctaagctc gctcggctgg gtcgcgggag
     7741 ggccggtcgc agtgattcag gcccttctgg attgtgttgg tccccagggc acgattgtca
     7801 tgcccacgca ctcgggtgat ctgactgatc ccgcagattg gagatcgccg cccgtgcctg
     7861 ccgattgggt gcagatccgt cgagttaacg cctcgactgt gccttctagt tgccagccat
     7921 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     7981 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     8041 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     8101 gggatgcggt gggctctatg ggatatcata acttcgtata gcatacatta tacgaagtta
     8161 tgctagcaat agaaactggg catgtggaga cagagaagac tcttgggttt ctgataggca
     8221 ctgactctct ctgcctattg gtctattttc ccacccttag gctgctggtg gtctaccctt
     8281 ggacccagag gttctttgag tcctttgggg atctgtccac tcctgatgct gttatgggca
     8341 accctaaggt gaaggctcat ggcaagaaag tgctcggtgc ctttagtgat ggcctggctc
     8401 acctggacaa cctcaagggc acctttgcca cactgagtga gctgcactgt gacaagctgc
     8461 acgtggatcc tgagaacttc agggtgagtc tatgggaccc ttgatgtttt ctttcccctt
     8521 cttttctatg gttaagttca tgtcatagga aggggagaag taacagggta cagtttagaa
     8581 tgggaaacag acgaatgatt gcatcagtgt ggaagtctca ggatcgtttt agtttctttt
     8641 atttgctgtt cataacaatt gttttctttt gtttaattct tgctttcttt ttttttcttc
     8701 tccgcaattt ttactattat acttaatgcc ttaacattgt gtataacaaa aggaaatatc
     8761 tctgagatac attaagtaac ttaaaaaaaa actttacaca gtctgcctag tacattacta
     8821 tttggaatat atgtgtgctt atttgcatat tcataatctc cctactttat tttcttttat
     8881 ttttaattga tacataatca ttatacatat ttatgggtta aagtgtaatg ttttaatatg
     8941 tgtacacata ttgaccaaat cagggtaatt ttgcatttgt aattttaaaa aatgctttct
     9001 tcttttaata tacttttttg tttatcttat ttctaatact ttccctaatc tctttctttc
     9061 agggcaataa tgatacaatg tatcatgcct ctttgcacca ttctaaagaa taacagtgat
     9121 aatttctggg ttaaggcaat agcaatattt ctgcatataa atatttctgc atataaattg
     9181 taactgatgt aagaggtttc atattgctaa tagcagctac aatccagcta ccattctgct
     9241 tttattttat ggttgggata aggctggatt attctgagtc caagctaggc ccttttgcta
     9301 atcatgttca tacctcttat cttcctccca cagctcctgg gcaacgtgct ggtctgtgtg
     9361 ctggcccatc actttggcaa agaattcacc ccaccagtgc aggctgccta tcagaaagtg
     9421 gtggctggtg tggctaatgc cctggcccac aagtatcact aagctcgctt tcttgctgtc
     9481 caatttctat taaaggttcc tttgttccct aagtccaact actaaactgg gggatattat
     9541 gaagggcctt gagcatctgg attctgccta ataaaaaaca tttattttca ttgcaatgat
     9601 gtatttaaat tatttctgaa tattttacta aaaagggaat gtgggaggtc agtgcattta
     9661 aaacataaag aaatgaagag ctagttcaaa ccttgggaaa atacactata tcttaaactc
     9721 catgaaagaa ggtgaggctg caaacagcta atgcacattg gcaacagccc ctgatgccta
     9781 tgccttattc atccctcaga aaaggattca agtagaggct tgatttggag gttaaagttt
     9841 tgctatgctg tattttacat tacttattgt tttagctgtc ctcatgaatg tcttttcact
     9901 acccatttgc ttatcctgca tctctcagcc ttgactccac tcagttctct tgcttagaga
     9961 taccaccttt cccctgaagt gttccttcca tgttttacgg cgagatggtt tctcctcgcc
    10021 tggccactca gccttagttg tctctgttgt cttatagagg tctacttgaa gaaggaaaaa
    10081 caggggtcat ggtttgactg tcctgtgagc ccttcttctc gacgataagc tttgcaaaga
    10141 tggataaagt tttaaacaga gaggaatctt tgcagctaat ggaccttcta ggtcttgaaa
    10201 ggagtgcctc gtgaggctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc
    10261 ccgagaagtt ggggggaggg gtcggcaatt gaaccggtgc ctagagaagg tggcgcgggg
    10321 taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac
    10381 cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa
    10441 cacaggtaag tgccgtgtgt ggttcccgcg ggcctggcct ctttacgggt tatggccctt
    10501 gcgtgccttg aattacttcc acctggctgc agtacgtgat tcttgatccc gagcttcggg
    10561 ttggaagtgg gtgggagagt tcgaggcctt gcgcttaagg agccccttcg cctcgtgctt
    10621 gagttgaggc ctggcctggg cgctggggcc gccgcgtgcg aatctggtgg caccttcgcg
    10681 cctgtctcgc tgctttcgat aagtctctag ccatttaaaa tttttgatga cctgctgcga
    10741 cgcttttttt ctggcaagat agtcttgtaa atgcgggcca agatctgcac actggtattt
    10801 cggtttttgg ggccgcgggc ggcgacgggg cccgtgcgtc ccagcgcaca tgttcggcga
    10861 ggcggggcct gcgagcgcgg ccaccgagaa tcggacgggg gtagtctcaa gctggccggc
    10921 ctgctctggt gcctggcctc gcgccgccgt gtatcgcccc gccctgggcg gcaaggctgg
    10981 cccggtcggc accagttgcg tgagcggaaa gatggccgct tcccggccct gctgcaggga
    11041 gctcaaaatg gaggacgcgg cgctcgggag agcgggcggg tgagtcaccc acacaaagga
    11101 aaagggcctt tccgtcctca gccgtcgctt catgtgactc cacggagtac cgggcgccgt
    11161 ccaggcacct cgattagttc tcgagctttt ggagtacgtc gtctttaggt tggggggagg
    11221 ggttttatgc gatggagttt ccccacactg agtgggtgga gactgaagtt aggccagctt
    11281 ggcacttgat gtaattctcc ttggaatttg ccctttttga gtttggatct tggttcattc
    11341 tcaagcctca gacagtggtt caaagttttt ttcttccatt tcaggtgtcg tgaggaatta
    11401 gcttggtact aatacgactc actataggga gacccaagct ggctagttaa gcttgatgct
    11461 accggactca gatccttggt ggcgtgaaac tcccgcacct ctttggcaag cgccttgtag
    11521 aagcgcgtat ggcttcgtac ccctgccatc aacacgcgtc tgcgttcgac caggctgcgc
    11581 gttctcgcgg ccatagcaac cgacgtacgg cgttgcgccc tcgccggcag caagaagcca
    11641 cggaagtccg cccggagcag aaaatgccca cgctactgcg ggtttatata gacggtcccc
    11701 acgggatggg gaaaaccacc accacgcaac tgctggtggc cctgggttcg cgcgacgata
    11761 tcgtctacgt acccgagccg atgacttact ggcgggtgct gggggcttcc gagacaatcg
    11821 cgaacatcta caccacacaa caccgcctcg accagggtga gatatcggcc ggggacgcgg
    11881 cggtggtaat gacaagcgcc cagataacaa tgggcatgcc ttatgccgtg accgacgccg
    11941 ttctggctcc tcatatcggg ggggaggctg ggagctcaca tgccccgccc ccggccctca
    12001 ccctcatctt cgaccgccat cccatcgccg ccctcctgtg ctacccggcc gcgcgatacc
    12061 ttatgggcag catgaccccc caggccgtgc tggcgttcgt ggccctcatc ccgccgacct
    12121 tgcccggcac aaacatcgtg ttgggggccc ttccggagga cagacacatc gaccgcctgg
    12181 ccaaacgcca gcgccccggc gagcggcttg acctggctat gctggccgcg attcgccgcg
    12241 tttacgggct gcttgccaat acggtgcggt atctgcaggg cggcgggtcg tggcgggagg
    12301 attggggaca gctttcgggg acggccgtgc cgccccaggg tgccgagccc cagagcaacg
    12361 cgggcccacg accccatatc ggggacacgt tatttaccct gtttcgggcc cccgagttgc
    12421 tggcccccaa cggcgacctg tataacgtgt ttgcctgggc cttggacgtc ttggccaaac
    12481 gcctccgtcc catgcacgtc tttatcctgg attacgacca atcgcccgcc ggctgccggg
    12541 acgccctgct gcaacttacc tccgggatgg tccagaccca cgtcaccacc ccaggctcca
    12601 taccgacgat ctgcgacctg gcgcgcacgt ttgcccggga tccaccggtc gccaccatgg
    12661 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
    12721 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
    12781 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
    12841 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
    12901 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
    12961 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
    13021 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
    13081 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
    13141 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
    13201 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
    13261 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
    13321 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaaagcg
    13381 gccgtacaag taaagcggcc gcgtcaacgc ctcgactgtg ccttctagtt gccagccatc
    13441 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
    13501 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
    13561 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
    13621 ggatgcggtg ggctctatgg gatgacgggc ccgcggaatt tcgagcatgc atctagaggg
    13681 cccaattcgc cctatagtga gtcgtattac aattcactgg ccgtcgtttt acaacgtcgt
    13741 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
    13801 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
    13861 aatggcgaat ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
    13921 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
    13981 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
    14041 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
    14101 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
    14161 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
    14221 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
    14281 aacaaaaatt taacgcgaat tttaacaaaa ttcagggcgc aagggctgct aaaggaagcg
    14341 gaacacgtag aaagccagtc cgcagaaacg gtgctgaccc cggatgaatg tcagctactg
    14401 ggctatctgg acaagggaaa acgcaagcgc aaagagaaag caggtagctt gcagtgggct
    14461 tacatggcga tagctagact gggcggtttt atggacagca agcgaaccgg aattgccagc
    14521 tggggcgccc tctggtaagg ttgggaagcc ctgcaaagta aactggatgg ctttcttgcc
    14581 gccaaggatc tgatggcgca ggggatcaag atctgatcaa gagacaggat gaggatcgtt
    14641 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
    14701 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
    14761 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
    14821 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
    14881 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
    14941 gcaggatctc ctgtcatccc accttgctcc tgccgagaaa gtatccatca tggctgatgc
    15001 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
    15061 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
    15121 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
    15181 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
    15241 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
    15301 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
    15361 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
    15421 tcttgacgag ttcttctgaa ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
    15481 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
    15541 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
    15601 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
    15661 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
    15721 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
    15781 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
    15841 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
    15901 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
    15961 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
    16021 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
    16081 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
    16141 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
    16201 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
    16261 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
    16321 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
    16381 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
    16441 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
    16501 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
    16561 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
    16621 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
    16681 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
    16741 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
    16801 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
    16861 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
    16921 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
    16981 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
    17041 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
    17101 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
    17161 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
    17221 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
    17281 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
    17341 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
    17401 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
    17461 a
//
