LOCUS       sequence-42185-e       15331 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15331)
  AUTHORS   Zariwala HA, Borghuis BG, Hoogland TM, Madisen L, Tian L, De Zeeuw 
            CI, Zeng H, Looger LL, Svoboda K, Chen TW
  TITLE     A Cre-dependent GCaMP3 reporter mouse for neuronal imaging in vivo.
  JOURNAL   J Neurosci. 2012 Feb 29;32(9):3131-41. doi: 
            10.1523/JNEUROSCI.4469-11.2012.
  PUBMED    22378886
REFERENCE   2  (bases 1 to 15331)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1523/JNEUROSCI.4469-11"; journalName: "J Neurosci"; date: 
            "2012-02-29- 29"; volume: "32"; issue: "9"; pages: "3131-41"
COMMENT     Sequence Label: Ai38
FEATURES             Location/Qualifiers
     source          1..15331
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    3..1087
                     /label=Rosa26 left arm
                     /note="left homology arm from intron 1 of the mouse Rosa26 
                     gene (Soriano, 1999)"
     enhancer        1181..1484
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1486..1763
                     /label=chicken beta-actin promoter
     intron          1765..2782
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2706..2728
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2790..2809
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    2858..2891
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     protein_bind    2915..2948
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    3116..3250
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    3370..3504
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    3624..3758
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    3787..3820
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      3940..3949
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3946..3954,3955..3972,3973..3975,3976..4008,4009..4011,
                     4012..4035,4036..4062,4063..4119,4120..4125,4126..4395,
                     4396..4413,4414..4848,4849..4851,4852..4854,4855..5298)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6f
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNTRDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     CDS             join(3946..3954,3955..3972,3973..3975,3976..4008,4009..4011,
                     4012..4035,4036..4062,4063..4119,4120..4125,4126..4395,
                     4396..4413,4414..4848,4849..4851,4852..4854,4855..5298)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6s
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNTRDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      4042..4051
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     4332..4353
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4459..4480)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4720..4739)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    5393..5981
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5446..5466)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5864..5875)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5984..6000)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5985..6001)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(5996..6013)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6002..6209
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    6217..6250
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     promoter        6272..6771
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(6307..6330)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     6704..6723
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     protein_bind    6791..6824
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     regulatory      6825..6834
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6831..7634
                     /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6894..6913)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7504..7523
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7644..8105
                     /label=PGK poly(A) signal
                     /note="mouse phosphoglycerate kinase 1 polyadenylation 
                     signal"
     misc_feature    8169..12438
                     /label=Rosa26 right arm
                     /note="right homology arm from intron 1 of the mouse Rosa26
                     gene (Soriano, 1999)"
     primer_bind     complement(12483..12503)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(12483..12501)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(12522..12538)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12522..12538)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12535..12557)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12546..12562
                     /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(12570..12576,12577..12594,12595..12600))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12615..12636
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12753..12770)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12924..13512)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13004..13023)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13683..14474,14475..14543))
                     /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     14306..14325
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14544..14648)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(14674..15129)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(14811..14832)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     15023..15042
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     15256..15278
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     15270..15287
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     15271..15287
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     15297..15316
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        15297..15315
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ccgcggcagg ccctccgagc gtggtggagc cgttctgtga gacagccggg tacgagtcgt
       61 gacgctggaa ggggcaagcg ggtggtgggc aggaatgcgg tccgccctgc agcaaccgga
      121 gggggaggga gaagggagcg gaaaagtctc caccggacgc ggccatggct cggggggggg
      181 ggggcagcgg aggascgctt ccggccgacg tctcgtcgct gattggctty ttttcctccc
      241 gccgtgtgtg aaaacacaaa tggcgtgttt tggttggcgt aaggcgcctg tcagttaacg
      301 gcagccggag tgcgcagccg ccggcagcct cgctctgccc actgggtggg gcgggaggta
      361 ggtggggtga ggcgagctgn acgtgcgggc gcggtcggcc tctggcgggg cgggggaggg
      421 gagggagggt cagcgaaagt agctcgcgcg cgagcggccg cccaccctcc ccttcctctg
      481 ggggagtcgt tttacccgcc gccggccggg cctcgtcgtc tgattggctc tcggggccca
      541 gaaaactggc ccttgccatt ggctcgtgtt cgtgcaagtt gagtccatcc gccggccagc
      601 gggggcggcg aggaggcgct cccaggttcc ggccctcccc tcggccccgc gccgcagagt
      661 ctggccgcgc gcccctgcgc aacgtggcag gaagcgcgcg ctgggggcgg ggacgggcag
      721 tagggctgag cggctgcggg gcgggtgcaa gcacgtttcc gacttgagtt gcctcaagag
      781 gggcgtgctg agccagacct ccatcgcgca ctccggggag tggagggaag gagcgagggc
      841 tcagttgggc tgttttggag gcaggaagca cttgctctcc caaagtcgct ctgagttgtt
      901 atcagtaagg gagctgcagt ggagtaggcg gggagaaggc cgcacccttc tccggagggg
      961 ggaggggagt gttgcaatac ctttctggga gttctctgct gcctcctggc ttctgaggac
     1021 cgccctgggc ctgggagaat cccttgcccc ctcttcccct cgtgatctgc aactccagtc
     1081 tttctagtta attaaccgtt taaacaattc tgcaggaatc tagttattaa tagtaatcaa
     1141 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     1201 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     1261 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     1321 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     1381 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     1441 ctacttggca gtacatctac gtattagtca tcgctattac catggtcgag gtgagcccca
     1501 cgttctgctt cactctcccc atctcccccc cctccccacc cccaattttg tatttattta
     1561 ttttttaatt attttgtgca gcgatggggg cggggggggg gggggggcgc gcgccaggcg
     1621 gggcggggcg gggcgagggg cggggcgggg cgaggcggag aggtgcggcg gcagccaatc
     1681 agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata
     1741 aaaagcgaag cgcgcggcgg gcggggagtc gctgcgacgc tgccttcgcc ccgtgccccg
     1801 ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg accgcgttac tcccacaggt
     1861 gagcgggcgg gacggccctt ctcctccggg ctgtaattag cgcttggttt aatgacggct
     1921 tgtttctttt ctgtggctgc gtgaaagcct tgaggggctc cgggagggcc ctttgtgcgg
     1981 ggggagcggc tcggggggtg cgtgcgtgtg tgtgtgcgtg gggagcgccg cgtgcggctc
     2041 cgcgctgccc ggcggctgtg agcgctgcgg gcgcggcgcg gggctttgtg cgctccgcag
     2101 tgtgcgcgag gggagcgcgg ccgggggcgg tgccccgcgg tgcggggggg gctgcgaggg
     2161 gaacaaaggc tgcgtgcggg gtgtgtgcgt gggggggtga gcagggggtg tgggcgcgtc
     2221 ggtcgggctg caaccccccc tgcacccccc tccccgagtt gctgagcacg gcccggcttc
     2281 gggtgcgggg ctccgtacgg ggcgtggcgc ggggctcgcc gtgccgggcg gggggtggcg
     2341 gcaggtgggg gtgccgggcg gggcggggcc gcctcgggcc ggggagggct cgggggaggg
     2401 gcgcggcggc ccccggagcg ccggcggctg tcgaggcgcg gcgagccgca gccattgcct
     2461 tttatggtaa tcgtgcgaga gggcgcaggg acttcctttg tcccaaatct gtgcggagcc
     2521 gaaatctggg aggcgccgcc gcaccccctc tagcgggcgc ggggcgaagc ggtgcggcgc
     2581 cggcaggaag gaaatgggcg gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct
     2641 ccctctccag cctcggggct gtccgcgggg ggacggctgc cttcgggggg gacggggcag
     2701 ggcggggttc ggcttctggc gtgtgaccgg cggctctaga gcctctgcta accatgttca
     2761 tgccttcttc tttttcctac agctcctggg caacgtgctg gttattgtgc tgtctcatca
     2821 ttttggcaaa gaattgattt gataccgcgg gccctaagaa gttcctattc tctagaaagt
     2881 ataggaactt cgtcgacatt taaatcattt aaatataact tcgtataatg tatgctatac
     2941 gaagttattc gcgatgaata aatgaaagct tgcagatctg cgactctaga ggatctgcga
     3001 ctctagagga tcataatcag ccntaccaca ttttgtagag gttttactng ctttaaaaaa
     3061 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     3121 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     3181 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     3241 tgtctggatc tgcgactcta gaggatcata atcagccata ccacatttgt agaggtttta
     3301 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     3361 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     3421 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     3481 aatgtatctt atcatgtctg gatctgcgac tctagaggat cataatcagc cataccacat
     3541 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     3601 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     3661 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     3721 tgtccaaact catcaatgta tcttatcatg tctggatccc catcaagctg atccggaacc
     3781 cttaatataa cttcgtataa tgtatgctat acgaagttat taggtccctc gacctgcagc
     3841 ccaagctaga tcgaattcgg ccggcccctt cacgtgttgc acttaacgcg ccaactagct
     3901 agctatcaag gttagtacgt tccaagcttg ggggaattcg ccaccatggg ttctcatcat
     3961 catcatcatc atggtatggc tagcatgact ggtggacagc aaatgggtcg ggatctgtac
     4021 gacgatgacg ataaggatct cgccaccatg gtcgactcat cacgtcgtaa gtggaataag
     4081 acaggtcacg cagtcagagc tataggtcgg ctgagctcac tcgagaacgt ctatatcaag
     4141 gccgacaagc agaagaacgg catcaaggcg aacttcaaga tccgccacaa catcgaggac
     4201 ggcggcgtgc agctcgccta ccactaccag cagaacaccc ccatcggcga cggccccgtg
     4261 ctgctgcccg acaaccacta cctgagcgtg cagtccaaac tttcgaaaga ccccaacgag
     4321 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     4381 gacgagctgt acaagggcgg taccggaggg agcatggtga gcaagggcga ggagctgttc
     4441 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     4501 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     4561 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     4621 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     4681 cccgaaggct acatccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     4741 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     4801 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacac gcgtgaccaa
     4861 ctgactgaag agcagatcgc agaatttaaa gaggctttct ccctatttga caaggacggg
     4921 gatgggacaa taacaaccaa ggagctgggg acggtgatgc ggtctctggg gcagaacccc
     4981 acagaagcag agctgcagga catgatcaat gaagtagatg ccgacggtga cggcacaatc
     5041 gacttccctg agttcctgac aatgatggca agaaaaatga aagacacaga cagtgaagaa
     5101 gaaattagag aagcgttccg tgtgtttgat aaggatggca atggctacat cagtgcagca
     5161 gagcttcgcc acgtgatgac aaaccttgga gagaagttaa cagatgaaga ggttgatgaa
     5221 atgatcaggg aagcagacat cgatggggat ggtcaggtaa actacgaaga gtttgtacaa
     5281 atgatgacag cgaagtaaga attccttggg ccccctcgaa ggtacgttgc gttatttaaa
     5341 tttggcgcgt acaaggccgg ccctgcagga attcgatatc aagcttatcg ataatcaacc
     5401 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     5461 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     5521 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     5581 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     5641 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     5701 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     5761 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt
     5821 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     5881 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     5941 ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc gtcgacctcg
     6001 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     6061 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     6121 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     6181 tgggaagaca atagcaggca tgctggggaa ctagtggtgc cagggcgtgc ccttgggctc
     6241 cccgggcgcg gcggccgcat cgaattctac cgggtagggg aggcgctttt cccaaggcag
     6301 tctggagcat gcgctttagc agccccgctg ggcacttggc gctacacaag tggcctctgg
     6361 cctcgcacac attccacatc caccggtagg cgccaaccgg ctccgttctt tggtggcccc
     6421 ttcgcgccac cttctactcc tcccctagtc aggaagttcc cccccgcccc gcagctcgcg
     6481 tcgtgcagga cgtgacaaat ggaagtagca cgtctcacta gtctcgtgca gatggacagc
     6541 accgctgagc aatggaagcg ggtaggcctt tggggcagcg gccaatagca gctttgctcc
     6601 ttcgctttct gggctcagag gctgggaagg ggtgggtccg ggggcgggct caggggcggg
     6661 ctcaggggcg gggcgggcgc ccgaaggtcc tccggaggcc cggcattctg cacgcttcaa
     6721 aagcgcacgt ctgccgcgct gttctcctct tcctcatctc cgggcctttc gacctgcaat
     6781 cgccgctagc gaagttccta ttctctagaa agtataggaa cttcgccacc atgggatcgg
     6841 ccattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     6901 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     6961 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     7021 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     7081 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     7141 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     7201 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     7261 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     7321 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     7381 gcgatgatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     7441 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     7501 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     7561 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     7621 acgagttctt ctgaggggat ccgctgtaag tctgcagaaa ttgatgatct attaaacaat
     7681 aaagatgtcc actaaaatgg aagtttttcc tgtcatactt tgttaagaag ggtgagaaca
     7741 gagtacctac attttgaatg gaaggattgg agctacgggg gtgggggtgg ggtgggatta
     7801 gataaatgcc tgctctttac tgaaggctct ttactattgc tttatgataa tgtttcatag
     7861 ttggatatca taatttaaac aagcaaaacc aaattaaggg ccagctcatt cctcccactc
     7921 atgatctata gatctataga tctctcgtgg gatcattgtt tttctcttga ttcccacttt
     7981 gtggttctaa gtactgtggt ttccaaatgt gtcagtttca tagcctgaag aacgagatca
     8041 gcagcctctg ttccacatac acttcattct cagtattgtt ttgccaagtt ctaattccat
     8101 cagaaagctt gcagaagatc tccccaactg gggtaacctt tgagttctct cagttggggg
     8161 ggcgcgccct agaagatggg cgggagtctt ctgggcaggc ttaaaggcta acctggtgtg
     8221 tgggcgttgt cctgcagggg aattgaacag gtgtaaaatt ggagggacaa gacttcccac
     8281 agattttcgg ttttgtcggg aagtttttta ataggggcaa ataggaaaat ggaggatagg
     8341 agtcatctgg ggtttatgca gcaaaactac aggtatattg cttgtatccg cctcggagat
     8401 ttccatgagg agataaagac atgtcacccg agtttatact ctcctgctta gatcctacta
     8461 cagtatgaaa tacagtgtyg cgaggtagac tatgtaagca gatttaatca ttttaaagag
     8521 cccagtactt catatccatt tctcccgctc cttctgcagc cttatcaaaa ggtatttaga
     8581 acactcattt tagccccatt ttcatttatt atactggctt atccaacccc tagacagagc
     8641 attggcattt tccctttcct gatcttagaa gtctgatgac tcatgaaacc agacagatta
     8701 gttacataca ccacaaatcg aggctgtagc tggggcctca acactgcagt tcttttataa
     8761 ctccttagta cactttttgt tgatcctttg ccttgatcct taattttcag tgtctatcac
     8821 ctctcccgtc aggtggtgtt ccacatttgg gcctattctc agtccaggga gttttacaac
     8881 aatagatgta ttgagaatcc aacctaaagc ttaactttcc actcccatga atgcctctct
     8941 cctttttctc cattataact gagctatwac cattaatggt ttcaggtgga tgtctcctcc
     9001 cccaatatac ctgatgtatc tacatattgc caggctgata ttttaagaca twaaaggtat
     9061 atttcattat tgagccacat ggtattgatt actgctacta aaattttgtc attgtacaca
     9121 tctgtaaaag gtggttcctt ttggaatgca aagttcaggt gtttgttgtc tttcctgacc
     9181 taaggtcttg tgagcttgta ttttttctat ttaagcagtg ctttctcttg gactggcttg
     9241 actcatggca ttctacacgt tattgctggt ctaaatgtga ttttgccaag cttcttcagg
     9301 acctataatt ttgcttgact tgtagccaaa cacaagtaaa atgattaagc aacaaatgta
     9361 tttgtgaagc ttggttttta ggttgttgtg ttgtgtgtgc ttgtgctcta taataatact
     9421 atccaggggc tggagaggtg gctcggagtt caagagcaca gactgctctt ccagaagtcc
     9481 tgagttcaat tcccagcaac cacatggtgg ctcacaacca tctgtaatgg gatctgatgc
     9541 cctcttctgg tgtgtctgaa gaccacaagt gtattcacat taaataaata atcctccttc
     9601 ttcttctttt ttttttttta aagagaatwc tgtctccagt agaattactg aagtaatgaa
     9661 atactttgtg tttgttccaa tatggwagcc aataatcaaa tactcttwag cactggaaat
     9721 gtaccaagga actattttat ttaagtgwac tgtggacaga ggagccataa ctgcagactt
     9781 gtgggataca gaagaccaat gcagacttaa tgtcttttct cttacactaa gcaataaaga
     9841 aataaaaatt gaacttctag tatcctattt gttaaactgc tagctttact aacttttgtg
     9901 cttcatctat acaaagctga aagctaagtc tgcagccatt actaaacatg aaagcaagta
     9961 atgataattt tggatttcaa aaatgtaggg ccagagttta gccagccagt ggtggtgctt
    10021 gcctttatgc cttaatccca gcactctgga ggcagagaca ggcagatctc tgagtttgag
    10081 cccagcctgg tctacacatc aagttctatc taggatagcc aggaatacac acagaaaccc
    10141 tgttggggag gggggctctg agatttcata aaattataat tgaagcattc cctaatgagc
    10201 cactatggat gtggctaaat ccgtctacct ttctgatgag atttgggtat tattttttct
    10261 gtctctgctg ttggttgggt cttttgacac tgtgggcttt cttaaagcct ccttccctgc
    10321 catgtggtct cttgtttgct actaacttcc catggcttaa atggcatggc tttttgcctt
    10381 ctaagggcag ctgctgagwt ttgcagcctg atttccaggg tggggttggg aaatctttca
    10441 aacactaaaa ttgtccttta attttttttt aaaaaatggg ttatataata aacctcataa
    10501 aatagttatg aggagtgagg tggactaata ttaatgagtc cctcccctat aaaagagcta
    10561 ttaaggcttt ttgtcttata ctaacttttt ttttaaatgt ggtatcttta gaaccaaggg
    10621 tcttagagtt ttagtataca gaaactgttg catcgcttaa tcagattttc tagtttcaaa
    10681 tccagagaat ccaaattctt cacagccaaa gtcaaattaa gaatttctga ctttaatgtt
    10741 atttgctact gtgaatataa aatgatagct tttcctgagg cagggtctca ctatgtatct
    10801 ctgcctgatc tgcaacaaga tatgtagact aaagttctgc ctgcttttgt ctcctgaata
    10861 ctaaggttaa aatgtagtaa tacttttgga acttgcaggt cagattcttt tataggggac
    10921 acactaaggg agcttgggtg atagttggta aatgtgttta agtgatgaaa acttgaatta
    10981 ttatcaccgc aacctacttt ttaaaaaaaa aagccaggcc tgttagagca tgctaaggga
    11041 tccctaggac ttgctgagca cacaagagta gtacttggca ggctcctggt gagagcatat
    11101 ttcaaaaaac aaggcagaca accaagaaac tacagtaagg ttacctgtct ttaaccatct
    11161 gcatatacac agggatatta aaatattcca aataatattt cattcaagtt ttcccccatc
    11221 aaattgggac atggatttct ccggtgaata ggcagagttg gaaactaaac aaatgttggt
    11281 tttgtgattt gtgaaattgt tttcaagtga tagttaaagc ccatgagata cagaacaaag
    11341 ctgctatttc gaggtctctt ggttatactc agaagcactt ctttgggttt ccctgcacta
    11401 tcctgatcat gtgctaggcc twccttaggc tgattgttgt tcaaataact taagtttcct
    11461 gtcaggtgat gtcatatgat ttcatatatc aaggcaaaac atgttatata tgttaaacat
    11521 ttgkacttaa tgtgaaagtt aggtctttgt gggttttgat tttaatttca aaacctgagc
    11581 taaataagtc attttacatg tcttacattt ggtgaattgt atattgtggt ttgcaggcaa
    11641 gactctctga cctagtaacc ctcctataga gcactttgct gggtcacaag tctaggagtc
    11701 aagcatttca ccttgaagtt gagacgtttt gttagtgtat actagttata tgttggagga
    11761 catgtttatc cagaagatat tcaggactat ttttgactgg gctaaggaat tgattctgat
    11821 tagcactgtt agtgagcatt gagtggcctt taggcttgaa ttggagtcac ttgtatatct
    11881 caaataatgc tggccttttt twaaaagccc ttgttcttta tcaccctgtt ttctacataa
    11941 tttttgttca aagaaatact tgtttggatc tccttttgac aacaatagca tgttttcaag
    12001 ccatattttt tttccttttt tttttttttt ttggtttttc gagacagggt ttctctgtat
    12061 agccctggct gtcctggaac tcactttgta gaccaggctg gcctcgaact cagaaatccg
    12121 cctgcctctg cctcctgagt gccgggatta aaggcgtgca ccaccacgcc tggctaagtt
    12181 ggatattttg tatataacta taaccaatac taactccact gggtggattt ttaattcagt
    12241 cagtagtctt aagtggtctt tattggccct tattaaaatc tactgttcac tctaacagag
    12301 gctgttggac tagtggsact aagcaacttc ctacggatat actagcagat aagggtcagg
    12361 gatagaaact agtctagcgt tttgtatacc taccagctta tactaccttg ttctgataga
    12421 aatatttagg acatctagct tatcgatccg tcgagggggg gcccggtacc cagcttttgt
    12481 tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg
    12541 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
    12601 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
    12661 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
    12721 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
    12781 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
    12841 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
    12901 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
    12961 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
    13021 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
    13081 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
    13141 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
    13201 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
    13261 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
    13321 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
    13381 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
    13441 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
    13501 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
    13561 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
    13621 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
    13681 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
    13741 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
    13801 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
    13861 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
    13921 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
    13981 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
    14041 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
    14101 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
    14161 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
    14221 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
    14281 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
    14341 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
    14401 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
    14461 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
    14521 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
    14581 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
    14641 gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa
    14701 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
    14761 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
    14821 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
    14881 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
    14941 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
    15001 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
    15061 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
    15121 agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
    15181 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
    15241 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat
    15301 acgactcact atagggcgaa ttggagctcc a
//
