pGPTVII-Bar-U-MatryoshCaMP6s

LOCUS       pGPTVII-Bar-U-MatryoshCaMP6s               13710 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Plant expression of fluorescent reporter for calcium signaling, 
            based off of GCaMP6s. Contains a stable reference Large Stokes Shift
            (LSS) mOrange nested within the reporting cpEGFP..
ACCESSION   .
VERSION     .
KEYWORDS    pGPTVII-Bar-U-MatryoshCaMP6s
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13710)
  AUTHORS   Ast C, Foret J, Oltrogge LM, De Michele R, Kleist TJ, Ho CH, Frommer
            WB
  TITLE     Ratiometric Matryoshka biosensors from a nested cassette of green- 
            and orange-emitting fluorescent proteins.
  JOURNAL   Nat Commun. 2017 Sep 5;8(1):431. doi: 10.1038/s41467-017-00400-2.
  PUBMED    28874729
REFERENCE   2  (bases 1 to 13710)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..13710
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             1..57
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     primer_bind     270..291
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             343..1050
                     /codon_start=1
                     /product="monomeric orange fluorescent protein with a large
                     Stokes shift (Shcherbakova et al., 2012)"
                     /label=LSSmOrange
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTVKLKVTKGGPLPFAWDILSPQFTYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGMEASSERMYPEDGALK
                     GEDKLRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     756..775
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1105..1126)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1366..1385)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2374..2398
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             2863..3513
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     primer_bind     complement(3701..3718)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3952..3971)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     oriT            4194..4303
                     /note="incP origin of transfer"
     CDS             4336..4707
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     rep_origin      4834..5545
                     /label=oriV
                     /note="incP origin of replication"
     mobile_element  6168..6935
                     /mobile_element_type="insertion sequence:IS1"
                     /note="prokaryotic transposable element"
     CDS             7127..7921
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     CDS             8220..9368
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     misc_feature    10964..10988
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     primer_bind     complement(11235..11256)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     11447..11466
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11591..11613
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     11605..11622
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     11606..11622
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      11635..11835
                     /label=gene 7 terminator
                     /note="transcription terminator for octopine-type Ti 
                     plasmid gene 7"
     CDS             complement(11870..12421)
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     promoter        complement(12472..12651)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
ORIGIN
        1 cgtcgtaagt ggaataagac aggtcacgca gtcagagcta taggtcggct gagctcactc
       61 gagaacgtct atatcaaggc cgacaagcag aagaacggca tcaaggcgaa cttccacatc
      121 cgccacaaca tcgaggacgg cggcgtgcag ctcgcctacc actaccagca gaacaccccc
      181 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcgtgca gtccaaactt
      241 tcgaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
      301 gggatcactc tcggcatgga cgagctgtac aagggcggta ccatggtgag caagggcgag
      361 gagaataaca tggccatcat caaggagttc atgcgcttca aggtgcgcat ggagggctcc
      421 gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcttt
      481 cagaccgtta agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcttg
      541 tcccctcagt tcacctacgg ctccaaggcc tacgtgaagc accccgccga catccccgac
      601 tacctcaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac
      661 ggcggcgtgg tgaccgtgac tcaggactcc tccctgcagg acggcgagtt catctacaag
      721 gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg
      781 ggcatggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgaggac
      841 aagctcaggc tgaagctgaa ggacggcggc cactacacct ccgaggtcaa gaccacctac
      901 aaggccaaga agcccgtgca gttgcccggc gcctacatcg tcgacatcaa gttggacatc
      961 acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac
     1021 tccaccggcg gcatggacga gctgtacaag ggagggagca tggtgagcaa gggcgaggag
     1081 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     1141 ttcagcgtgt ccggcgaggg tgagggcgat gccacctacg gcaagctgac cctgaagttc
     1201 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     1261 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     1321 gccatgcccg aaggctacat ccaggagcgc accatcttct tcaaggacga cggcaactac
     1381 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     1441 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caacctgccg
     1501 gaccaactga ctgaagagca gatcgcagaa tttaaagagg ctttctccct atttgacaag
     1561 gacggggatg ggacaataac aaccaaggag ctggggacgg tgatgcggtc tctggggcag
     1621 aaccccacag aagcagagct gcaggacatg atcaatgaag tagatgccga cggtgacggc
     1681 acaatcgact tccctgagtt cctgacaatg atggcaagaa aaatgaaata cagggacacg
     1741 gaagaagaaa ttagagaagc gttcggtgtg tttgataagg atggcaatgg ctacatcagt
     1801 gcagcagagc ttcgccacgt gatgacaaac cttggagaga agttaacaga tgaagaggtt
     1861 gatgaaatga tcagggaagc agacatcgat ggggatggtc aggtaaacta cgaagagttt
     1921 gtacaaatga tgacagcgaa gtgacccggg taggagctca tatgaagatg aagatgaaat
     1981 atttggtgtg tcaaataaat agcttgtgtg cttaagtttg tgtttttttc ttggcttgtt
     2041 gtgttatgaa tttgtggctt tttctaatat taaatgaatg taagatctca ttataatgaa
     2101 taaacaaatg tttctataat ccattgtgaa tgttttgttg gatctcttct gcagcatata
     2161 actactgtat gtgctatggt atggactatg gaatatgatt aaagataagg aattcagatc
     2221 ggctgagtgg ctccttcaac gttgcggttc tgtcagttcc aaacgtaaaa cggcttgtcc
     2281 cgcgtcatcg gcgggggtca taacgtgact cccttaattc tccgctcatg atcagattgt
     2341 cgtttcccgc cttcagttta aactatcagt gtttgacagg atatattggc gggtaaacct
     2401 aagagaaaag agcgtttatt agaataatcg gatatttaaa agggcgtgaa aaggtttatc
     2461 cgttcgtcca tttgtatgtg catgccaacc acagggttcc ccagatctgg cgccggccag
     2521 cgagacgagc aagattggcc gccgcccgaa acgatccgac agcgcgccca gcacaggtgc
     2581 gcaggcaaat tgcaccaacg catacagcgc cagcagaatg ccatagtggg cggtgacgtc
     2641 gttcgagtga accagatcgc gcaggaggcc cggcagcacc ggcataatca ggccgatgcc
     2701 gacagcgtcg agcgcgacag tgctcagaat tacgatcagg ggtatgttgg gtttcacgtc
     2761 tggcctccgg accagcctcc gctggtccga ttgaacgcgc ggattcttta tcactgataa
     2821 gttggtggac atattatgtt tatcagtgat aaagtgtcaa gcatgacaaa gttgcagccg
     2881 aatacagtga tccgtgccgc cctggacctg ttgaacgagg tcggcgtaga cggtctgacg
     2941 acacgcaaac tggcggaacg gttgggggtt cagcagccgg cgctttactg gcacttcagg
     3001 aacaagcggg cgctgctcga cgcactggcc gaagccatgc tggcggagaa tcatacgcat
     3061 tcggtgccga gagccgacga cgactggcgc tcatttctga tcgggaatgc ccgcagcttc
     3121 aggcaggcgc tgctcgccta ccgcgatggc gcgcgcatcc atgccggcac gcgaccgggc
     3181 gcaccgcaga tggaaacggc cgacgcgcag cttcgcttcc tctgcgaggc gggtttttcg
     3241 gccggggacg ccgtcaatgc gctgatgaca atcagctact tcactgttgg ggccgtgctt
     3301 gaggagcagg ccggcgacag cgatgccggc gagcgcggcg gcaccgttga acaggctccg
     3361 ctctcgccgc tgttgcgggc cgcgatagac gccttcgacg aagccggtcc ggacgcagcg
     3421 ttcgagcagg gactcgcggt gattgtcgat ggattggcga aaaggaggct cgttgtcagg
     3481 aacgttgaag gaccgagaaa gggtgacgat tgatcaggac cgctgccgga gcgcaaccca
     3541 ctcactacag cagagccatg tagacaacat cccctccccc tttccaccgc gtcagacgcc
     3601 cgtagcagcc cgctacgggc tttttcatgc cctgccctag cgtccaagcc tcacggccgc
     3661 gctcggcctc tctggcggcc ttctggcgct cttccgcttc ctcgctcact gactcgctgc
     3721 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     3781 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     3841 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3901 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3961 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4021 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct tttccgctgc ataaccctgc
     4081 ttcggggtca ttatagcgat tttttcggta tatccatcct ttttcgcacg atatacagga
     4141 ttttgccaaa gggttcgtgt agactttcct tggtgtatcc aacggcgtca gccgggcagg
     4201 ataggtgaag taggcccacc cgcgagcggg tgttccttct tcactgtccc ttattcgcac
     4261 ctggcggtgc tcaacgggaa tcctgctctg cgaggctggc cggctaccgc cggcgtaaca
     4321 gatgagggca agcggatggc tgatgaaacc aagccaacca ggaagggcag cccacctatc
     4381 aaggtgtact gccttccaga cgaacgaaga gcgattgagg aaaaggcggc ggcggccggc
     4441 atgagcctgt cggcctacct gctggccgtc ggccagggct acaaaatcac gggcgtcgtg
     4501 gactatgagc acgtccgcga gctggcccgc atcaatggcg acctgggccg cctgggcggc
     4561 ctgctgaaac tctggctcac cgacgacccg cgcacggcgc ggttcggtga tgccacgatc
     4621 ctcgccctgc tggcgaagat cgaagagaag caggacgagc ttggcaaggt catgatgggc
     4681 gtggtccgcc cgagggcaga gccatgactt ttttagccgc taaaacggcc ggggggtgcg
     4741 cgtgattgcc aagcacgtcc ccatgcgctc catcaagaag agcgacttcg cggagctggt
     4801 gaagtacatc accgacgagc aaggcaagac cgagcgcctt tccgacgctc accgggctgg
     4861 ttgccctcgc cgctgggctg gcggccgtct atggccctgc aaacgcgcca gaaacgccgt
     4921 cgaagccgtg tgcgagacac cgcggccgcc ggcgttgtgg atacctcgcg gaaaacttgg
     4981 ccctcactga cagatgaggg gcggacgttg acacttgagg ggccgactca cccggcgcgg
     5041 cgttgacaga tgaggggcag gctcgatttc ggccggcgac gtggagctgg ccagcctcgc
     5101 aaatcggcga aaacgcctga ttttacgcga gtttcccaca gatgatgtgg acaagcctgg
     5161 ggataagtgc cctgcggtat tgacacttga ggggcgcgac tactgacaga tgaggggcgc
     5221 gatccttgac acttgagggg cagagtgctg acagatgagg ggcgcaccta ttgacatttg
     5281 aggggctgtc cacaggcaga aaatccagca tttgcaaggg tttccgcccg tttttcggcc
     5341 accgctaacc tgtcttttaa cctgctttta aaccaatatt tataaacctt gtttttaacc
     5401 agggctgcgc cctgtgcgcg tgaccgcgca cgccgaaggg gggtgccccc ccttctcgaa
     5461 ccctcccggc ccgctaacgc gggcctccca tccccccagg ggctgcgccc ctcggccgcg
     5521 aacggcctca ccccaaaaat ggcagcgctg gcagtccttg ccattgccgg gatcggggca
     5581 gtaacgggat gggcgatcag cccgagcgcg acgcccggaa gcattgacgt gccgcaggtg
     5641 ctggcatcga cattcagcga ccaggtgccg ggcagtgagg gcggcggcct gggtggcggc
     5701 ctgcccttca cttcggccgt cggggcattc acggacttca tggcggggcc ggcaattttt
     5761 accttgggca ttcttggcat agtggtcgcg ggtgccgtgc tcgtgttcgg gggtgcgata
     5821 aacccagcga accatttgag gtgataggta agattatacc gaggtatgaa aacgagaatt
     5881 ggacctttac agaattactc tatgaagcgc catatttaaa aagctaccaa gacgaagagg
     5941 atgaagagga tgaggaggca gattgccttg aatatattga caatactgat aagataatat
     6001 atcttttata tagaagatat cgccgtatgt aaggatttca gggggcaagg cataggcagc
     6061 gcgcttatca atatatctat agaatgggca aagcataaaa acttgcatgg actaatgctt
     6121 gaaacccagg acaataacct tatagcttgt aaattctatc ataattgggt aatgactcca
     6181 acttattgat agtgttttat gttcagataa tgcccgatga ctttgtcatg cagctccacc
     6241 gattttgaga acgacagcga cttccgtccc agccgtgcca ggtgctgcct cagattcagg
     6301 ttatgccgct caattcgctg cgtatatcgc ttgctgatta cgtgcagctt tcccttcagg
     6361 cgggattcat acagcggcca gccatccgtc atccatatca ccacgtcaaa gggtgacagc
     6421 aggctcataa gacgccccag cgtcgccata gtgcgttcac cgaatacgtg cgcaacaacc
     6481 gtcttccgga gactgtcata cgcgtaaaac agccagcgct ggcgcgattt agccccgaca
     6541 tagccccact gttcgtccat ttccgcgcag acgatgacgt cactgcccgg ctgtatgcgc
     6601 gaggttaccg actgcggcct gagtttttta agtgacgtaa aatcgtgttg aggccaacgc
     6661 ccataatgcg ggctgttgcc cggcatccaa cgccattcat ggccatatca atgattttct
     6721 ggtgcgtacc gggttgagaa gcggtgtaag tgaactgcag ttgccatgtt ttacggcagt
     6781 gagagcagag atagcgctga tgtccggcgg tgcttttgcc gttacgcacc accccgtcag
     6841 tagctgaaca ggagggacag ctgatagaaa cagaagccac tggagcacct caaaaacacc
     6901 atcatacact aaatcagtaa gttggcagca tcacccataa ttgtggtttc aaaatcggct
     6961 ccgtcgatac tatgttatac gccaactttg aaaacaactt tgaaaaagct gttttctggt
     7021 atttaaggtt ttagaatgca aggaacagtg aattggagtt cgtcttgtta taattagctt
     7081 cttggggtat ctttaaatac tgtagaaaag aggaaggaaa taataaatgg ctaaaatgag
     7141 aatatcaccg gaattgaaaa aactgatcga aaaataccgc tgcgtaaaag atacggaagg
     7201 aatgtctcct gctaaggtat ataagctggt gggagaaaat gaaaacctat atttaaaaat
     7261 gacggacagc cggtataaag ggaccaccta tgatgtggaa cgggaaaagg acatgatgct
     7321 atggctggaa ggaaagctgc ctgttccaaa ggtcctgcac tttgaacggc atgatggctg
     7381 gagcaatctg ctcatgagtg aggccgatgg cgtcctttgc tcggaagagt atgaagatga
     7441 acaaagccct gaaaagatta tcgagctgta tgcggagtgc atcaggctct ttcactccat
     7501 cgacatatcg gattgtccct atacgaatag cttagacagc cgcttagccg aattggatta
     7561 cttactgaat aacgatctgg ccgatgtgga ttgcgaaaac tgggaagaag acactccatt
     7621 taaagatccg cgcgagctgt atgatttttt aaagacggaa aagcccgaag aggaacttgt
     7681 cttttcccac ggcgacctgg gagacagcaa catctttgtg aaagatggca aagtaagtgg
     7741 ctttattgat cttgggagaa gcggcagggc ggacaagtgg tatgacattg ccttctgcgt
     7801 ccggtcgatc agggaggata tcggggaaga acagtatgtc gagctatttt ttgacttact
     7861 ggggatcaag cctgattggg agaaaataaa atattatatt ttactggatg aattgtttta
     7921 gtacctagat gtggcgcaac gatgccggcg acaagcagga gcgcaccgac ttcttccgca
     7981 tcaagtgttt tggctctcag gccgaggccc acggcaagta tttgggcaag gggtcgctgg
     8041 tattcgtgca gggcaagatt cggaatacca agtacgagaa ggacggccag acggtctacg
     8101 ggaccgactt cattgccgat aaggtggatt atctggacac caaggcacca ggcgggtcaa
     8161 atcaggaata agggcacatt gccccggcgt gagtcggggc aatcccgcaa ggagggtgaa
     8221 tgaatcggac gtttgaccgg aaggcataca ggcaagaact gatcgacgcg gggttttccg
     8281 ccgaggatgc cgaaaccatc gcaagccgca ccgtcatgcg tgcgccccgc gaaaccttcc
     8341 agtccgtcgg ctcgatggtc cagcaagcta cggccaagat cgagcgcgac agcgtgcaac
     8401 tggctccccc tgccctgccc gcgccatcgg ccgccgtgga gcgttcgcgt cgtctcgaac
     8461 aggaggcggc aggtttggcg aagtcgatga ccatcgacac gcgaggaact atgacgacca
     8521 agaagcgaaa aaccgccggc gaggacctgg caaaacaggt cagcgaggcc aagcaggccg
     8581 cgttgctgaa acacacgaag cagcagatca aggaaatgca gctttccttg ttcgatattg
     8641 cgccgtggcc ggacacgatg cgagcgatgc caaacgacac ggcccgctct gccctgttca
     8701 ccacgcgcaa caagaaaatc ccgcgcgagg cgctgcaaaa caaggtcatt ttccacgtca
     8761 acaaggacgt gaagatcacc tacaccggcg tcgagctgcg ggccgacgat gacgaactgg
     8821 tgtggcagca ggtgttggag tacgcgaagc gcacccctat cggcgagccg atcaccttca
     8881 cgttctacga gctttgccag gacctgggct ggtcgatcaa tggccggtat tacacgaagg
     8941 ccgaggaatg cctgtcgcgc ctacaggcga cggcgatggg cttcacgtcc gaccgcgttg
     9001 ggcacctgga atcggtgtcg ctgctgcacc gcttccgcgt cctggaccgt ggcaagaaaa
     9061 cgtcccgttg ccaggtcctg atcgacgagg aaatcgtcgt gctgtttgct ggcgaccact
     9121 acacgaaatt catatgggag aagtaccgca agctgtcgcc gacggcccga cggatgttcg
     9181 actatttcag ctcgcaccgg gagccgtacc cgctcaagct ggaaaccttc cgcctcatgt
     9241 gcggatcgga ttccacccgc gtgaagaagt ggcgcgagca ggtcggcgaa gcctgcgaag
     9301 agttgcgagg cagcggcctg gtggaacacg cctgggtcaa tgatgacctg gtgcattgca
     9361 aacgctaggg ccttgtgggg tcagttccgg ctgggggttc agcagccagc gctttactgg
     9421 catttcagga acaagcgggc actgctcgac gcacttgctt cgctcagtat cgctcgggac
     9481 gcacggcgcg ctctacgaac tgccgataaa cagaggatta aaattgacaa ttgtgattaa
     9541 ggctcagatt cgacggcttg gagcggccga cgtgcaggat ttccgcgaga tccgattgtc
     9601 ggccctgaag aaagctccag agatgttcgg gtccgtttac gagcacgagg agaaaaagcc
     9661 catggaggcg ttcgctgaac ggttgcgaga tgccgtggca ttcggcgcct acatcgacgg
     9721 cgagatcatt gggctgtcgg tcttcaaaca ggaggacggc cccaaggacg ctcacaaggc
     9781 gcatctgtcc ggcgttttcg tggagcccga acagcgaggc cgaggggtcg ccggtatgct
     9841 gctgcgggcg ttgccggcgg gtttattgct cgtgatgatc gtccgacaga ttccaacggg
     9901 aatctggtgg atgcgcatct tcatcctcgg cgcacttaat atttcgctat tctggagctt
     9961 gttgtttatt tcggtctacc gcctgccggg cggggtcgcg gcgacggtag gcgctgtgca
    10021 gccgctgatg gtcgtgttca tctctgccgc tctgctaggt agcccgatac gattgatggc
    10081 ggtcctgggg gctatttgcg gaactgcggg cgtggcgctg ttggtgttga caccaaacgc
    10141 agcgctagat cctgtcggcg tcgcagcggg cctggcgggg gcggtttcca tggcgttcgg
    10201 aaccgtgctg acccgcaagt ggcaacctcc cgtgcctctg ctcaccttta ccgcctggca
    10261 actggcggcc ggaggacttc tgctcgttcc agtagcttta gtgtttgatc cgccaatccc
    10321 gatgcctaca ggaaccaatg ttctcggcct ggcgtggctc ggcctgatcg gagcgggttt
    10381 aacctacttc ctttggttcc gggggatctc gcgactcgaa cctacagttg tttccttact
    10441 gggctttctc agccccagat ctggggtcga tcagccgggg atgcatcagg ccgacagtcg
    10501 gaacttcggg tccccgacct gtaccattcg gtgagcaatg gataggggag ttgatatcgt
    10561 caacgttcac ttctaaagaa atagcgccac tcagcttcct cagcggcttt atccagcgat
    10621 ttcctattat gtcggcatag ttctcaagat cgacagcctg tcacggttaa gcgagaaatg
    10681 aataagaagg ctgataattc ggatctctgc gagggagatg atatttgatc acaggcagca
    10741 acgctctgtc atcgttacaa tcaacatgct accctccgcg agatcatccg tgtttcaaac
    10801 ccggcagctt agttgccgtt cttccgaata gcatcggtaa catgagcaaa gtctgccgcc
    10861 ttacaacggc tctcccgctg acgccgtccc ggactgatgg gctgcctgta tcgagtggtg
    10921 attttgtgcc gagctgccgg tcggggagct gttggctggc tggtggcagg atatattgtg
    10981 gtgtaaacaa attgacgctt agacaactta ataacacatt gcggacgttt ttaatgtact
    11041 ggggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg
    11101 ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt
    11161 ttgatggtgg ttccgaaatc ggcaaaatcc cttataaatc aaaagaatag cccgagatag
    11221 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
    11281 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcacccaaat
    11341 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
    11401 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
    11461 aaggagcggg cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
    11521 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
    11581 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattaat tcccatcttg
    11641 aaagaaatat agtttaaata tttattgata aaataacaag tcaggtatta tagtccaagc
    11701 aaaaacataa atttattgat gcaagtttaa attcagaaat atttcaataa ctgattatat
    11761 cagctggtac attgccgtag atgaaagact gagtgcgata ttatgtgtaa tacataaatt
    11821 gatgatatag ctagcttagc tcatcggggg atcgatcctc taggggtcat cagatctcgg
    11881 tgacgggcag gaccggacgg ggcggtaccg gcaggctgaa gtccagctgc cagaaaccca
    11941 cgtcatgcca gttcccgtgc ttgaagccgg ccgcccgcag catgccgcgg ggggcatatc
    12001 cgagcgcctc gtgcatgcgc acgctcgggt cgttgggcag cccgatgaca gcgaccacgc
    12061 tcttgaagcc ctgtgcctcc agggacttca gcaggtgggt gtagagcgtg gagcccagtc
    12121 ccgtccgctg gtggcggggg gagacgtaca cggtcgactc ggccgtccag tcgtaggcgt
    12181 tgcgtgcctt ccaggggccc gcgtaggcga tgccggcgac ctcgccgtcc acctcggcga
    12241 cgagccaggg atagcgctcc cgcagacgga cgaggtcgtc cgtccactcc tgcggttcct
    12301 gcggctcggt acggaagttg accgtgcttg tctcgatgta gtggttgacg atggtgcaga
    12361 ccgccggcat gtccgcctcg gtggcacggc ggatgtcggc cgggcgtcgt tctgggctca
    12421 tggtagatcc ccctagagtc gagatctgga ttgagagtga atatgagact ctaattggat
    12481 accgagggga atttatggaa cgtcagtgga gcatttttga caagaaatat ttgctagctg
    12541 atagtgacct taggcgactt ttgaacgcgc aataatggtt tctgacgtat gtgcttagct
    12601 cattaaactc cagaaacccg cggctgagtg gctccttcaa cgttgcggtt ctgtcagttc
    12661 caaacgtaaa acggcttgtc ccgcgtcatc ggcgggggtc ataacgtgac tcccttaatt
    12721 ctccgctcat gatcttgatc ccctgcgcca tcagatcctt ggcggcaaga aagccatcca
    12781 gtttactttg cagggcttcc caaccttacc agagggcgcc ccagctggca attccggttc
    12841 gcttgctgtc cataaaaccg cccagtctag ctatcgccat gtaagcccac tgcaagctac
    12901 ctgctttctc tttgcgcttg cgttttccct tgtccagata gcccagtagc tgacattcat
    12961 ccggggtcag caccgtttct gcggactggc tttctacgtg ttccgcttcc tttagcagcc
    13021 cttgcgccct gagtgcttgc ggcagcgtga agcttcgacg agtcagtaat aaacggcgtc
    13081 aaagtggttg cagccggcac acacgagtcg tgtttatcaa ctcaaagcac aaatactttt
    13141 cctcaaccta aaaataaggc aattagccaa aaacaacttt gcgtgtaaac aacgctcaat
    13201 acacgtgtca ttttattatt agctattgct tcaccgcctt agctttctcg tgacctagtc
    13261 gtcctcgtct tttcttcttc ttcttctata aaacaatacc caaagacctc ttcttcttca
    13321 caattcagat ttcaatttct caaaatctta aaaactttct ctcaattctc tctaccgtga
    13381 tcaaggtaaa tttctgtgtt ccttattctc tcaaaatctt cgattttgtt ttcgttcgat
    13441 cccaatttcg tatatgttct ttggtttaga ttctgttaat cttagatcga agacgatttt
    13501 ctgggtttga tcgttagata tcatcttaat tctcgattag ggtttcatag atatcatccg
    13561 atttgttcaa ataatttgag ttttgtcgaa taattactct tcgatttgtg atttctatct
    13621 agatctggtg ttagtttcta gtttgtgcga tcgaatttgt cgattaatct gagtttttct
    13681 gattaacaga ctagtatggt cgactcatca
//