LOCUS       sequence-460683-       19547 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bxb1-specific donor plasmid for integrating transcription units for 
            expressing ASAP2f (2 copies), jRCaMP1b and membrane-localised 
            miRFP703.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 19547)
  AUTHORS   Blanch-Asensio A, Grandela C, Brandao KO, de Korte T, Mei H, 
            Ariyurek Y, Yiangou L, Mol MPH, van Meer BJ, Kloet SL, Mummery CL, 
            Davis RP
  TITLE     STRAIGHT-IN enables high-throughput targeting of large DNA payloads 
            in human pluripotent stem cells.
  JOURNAL   Cell Rep Methods. 2022 Sep 22;2(10):100300. doi: 
            10.1016/j.crmeth.2022.100300. eCollection 2022 Oct 24.
  PUBMED    36313798
REFERENCE   2  (bases 1 to 19547)
  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: "Cell Rep 
            Methods."; date: "2022-09-22"; volume: "2(10):100300. doi"; pages: "
            10.1016/j.crmeth.2022.100300. eCollection 2022 Oct 24"
COMMENT     Sequence Label: Bxb1_AJMA
FEATURES             Location/Qualifiers
     source          1..19547
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    51..75
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        151..1329
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          382..1320
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1277..1297
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1337..1346
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    1349..1386
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attB site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     protein_bind    complement(1395..1428)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     enhancer        1530..1833
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1835..2110
                     /label=chicken beta-actin promoter
     intron          2111..3127
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     regulatory      3189..3198
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4626..4681
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     enhancer        5089..5392
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          5669..6685
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     CDS             6819..6836
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             6840..6872
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(6876..6884,6885..6899)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     polyA_signal    8289..8344
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     enhancer        8752..9055
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          9333..10350
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     regulatory      10412..10421
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             10463..11410
                     /codon_start=1
                     /product="bright monomeric near-infrared fluorescent 
                     protein engineered from a bacterial phytochrome, with an 
                     emission maximum of 703 nm (Shcherbakova et al., 2016)"
                     /label=miRFP703
                     /label=ORF frame 1
                     /translation="MVAGHASGSPAFGTASHSNCEHEEIHLAGSIQPHGALLVVSEHDH
                     RVIQASANAAEFLNLGSVLGVPLAEIDGDLLIKILPHLDPTAEGMPVAVRCRIGNPSTE
                     YCGLMHRPPEGGLIIELERAGPSIDLSGTLAPALERIRTAGSLRALCDDTVLLFQQCTG
                     YDRVMVYRFDEQGHGLVFSECHVPGLESYFGNRYPSSLVPQMARQLYVRQRVRVLVDVT
                     YQPVPLEPRLSPLTGRDLDMSGCFLRSMSPIHLQFLKDMGVRATLAVSLVVGGKLWGLV
                     VCHHYLPRFIRFELRAICKRLAERIATRITALES"
     polyA_signal    11532..11653
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11569..11588)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11623..11642
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     enhancer        11835..12138
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          12416..13432
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     regulatory      13494..13503
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    14931..14986
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     protein_bind    complement(15374..15406)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     primer_bind     complement(15414..15431)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(15414..15430)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    15467..15491
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      15579..16289
                     /label=oriV
                     /note="incP origin of replication"
     promoter        16310..16414
                     /gene="bla"
                     /label=AmpR promoter
     CDS             16415..17209
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      17380..17968
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     17869..17888
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             18304..19452
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
ORIGIN
        1 agtggtgatt ttgtgccgag ctgccggtcg gggagctgtt ggctggctgg tggcaggata
       61 tattgtggtg taaacaaatt gacgcttaga caacttaata acacattgcg gacgttttta
      121 atgtactggg gttgaacact ctgtgccatt ggctccggtg cccgtcagtg ggcagagcgc
      181 acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac cggtgcctag
      241 agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc
      301 gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac
      361 gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt cccgcgggcc tggcctcttt
      421 acgggttatg gcccttgcgt gccttgaatt acttccacct ggctgcagta cgtgattctt
      481 gatcccgagc ttcgggttgg aagtgggtgg gagagttcga ggccttgcgc ttaaggagcc
      541 ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct ggggccgccg cgtgcgaatc
      601 tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt ctctagccat ttaaaatttt
      661 tgatgacctg ctgcgacgct ttttttctgg caagatagtc ttgtaaatgc gggccaagat
      721 ctgcacactg gtatttcggt ttttggggcc gcgggcggcg acggggcccg tgcgtcccag
      781 cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac cgagaatcgg acgggggtag
      841 tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc cgccgtgtat cgccccgccc
      901 tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag cggaaagatg gccgcttccc
      961 ggccctgctg cagggagctc aaaatggagg acgcggcgct cgggagagcg ggcgggtgag
     1021 tcacccacac aaaggaaaag ggcctttccg tcctcagccg tcgcttcatg tgactccacg
     1081 gagtaccggg cgccgtccag gcacctcgat tagttctcga gcttttggag tacgtcgtct
     1141 ttaggttggg gggaggggtt ttatgcgatg gagtttcccc acactgagtg ggtggagact
     1201 gaagttaggc cagcttggca cttgatgtaa ttctccttgg aatttgccct ttttgagttt
     1261 ggatcttggt tcattctcaa gcctcagaca gtggttcaaa gtttttttct tccatttcag
     1321 gtgtcgtgag gaattcgcca ccatggccgg cttgtcgacg acggcggtct ccgtcgtcag
     1381 gatcatccgg atccataact tcgtatagca tacattatac gaagttatca tgatattcgg
     1441 caagcaggca tcgtagctgc caattcggat ccggagacta actacggtga aagagactta
     1501 gaaggagatt gagagatact ggagttccgc gttacataac ttacggtaaa tggcccgcct
     1561 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
     1621 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     1681 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
     1741 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
     1801 tacatctacg tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc
     1861 actctcccca tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta
     1921 ttttgtgcag cgatgggggc gggggggggg ggggcgcgcg ccrggcgggg cggggcgggg
     1981 gggggggggg gggggggcgr ggcggrgagg tgcggcggca gccaatcaga gcggcgcgct
     2041 ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc
     2101 gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc
     2161 gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg
     2221 cccttctcct ccgggctgta attagcgctt ggtttaatga cggcttgttt cttttctgtg
     2281 gctgcgtgaa agccttgagg ggctccggga gggccctttg tgcgggggga gcggctcggg
     2341 gggtgcgtgc gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg
     2401 ctgtgagcgc tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag
     2461 cgcggccggg ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt
     2521 gcggggtgtg tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc
     2581 ccccctgcac ccccctcccc gagttgctga gcacggcccg gcttcgggtg cggggctccg
     2641 tacggggcgt ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc
     2701 gggcggggcg gggccgcctc gggccggggr gggctcgggg grggggcgcg gcggcccccg
     2761 gagcgccggc ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg
     2821 cgagagggcg cagggacttc ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg
     2881 ccgccgcacc ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat
     2941 gggcggggag ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg
     3001 gggctgtccg cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt
     3061 ctggcgtgtg accggcggct ctagagcctc tgctaaccat gttcatgcct tcttcttttt
     3121 cctacagctc ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt
     3181 ccgatcgcgc caccatggag acgactgtga ggtatgaaca ggggtcagag ctcactaaaa
     3241 cttcgagctc tccaacagca gatgagccca cgataaagat tgatgatggt cgtgatgagg
     3301 gtaatgaaca agacagctgt tccaatacca ttaggagaaa aatttccccg tttgtgatgt
     3361 catttggatt cagagtattt ggagttgtgc ttatcattgt agacatcata gtggtgattg
     3421 tggatctggc catcagtgag aagaaaagag gcattagaga gattcttgaa ggtgtttccc
     3481 tggctatagc actcttcttc cttgttgatg ttctcatgag agtgtttgtt gaaggcttca
     3541 agaactattt ccggtccaaa ctgaatactt tggatgcagt catagtagtg ggcactctgc
     3601 taattaatat gacctactcc ttctctgacc ttagctttaa cagccataac gtgtatatta
     3661 ccgcggataa acagaaaaac ggcattaaag cgaactttac cgtgcgccat aacgtggaag
     3721 atggcagcgt gcagctggcg gatcattatc agcagaacac cccgattggc gatggcccgg
     3781 tgctgctgcc ggataaccat tatctgagca cccagaccgt gctgagcaaa gatccgaacg
     3841 aaaaacgcga tcacatggtg ctgctggaat ttgtgaccgc agcgggcatt acacacggca
     3901 tggatgaact gtatggcggc accggcggca gcgcgagcca gggcgaagaa ctgtttaccg
     3961 gcgtggtgcc gattctggtg gaactggatg gcgatgtgaa cggccataaa tttagcgtgc
     4021 gcggcgaagg cgaaggcgat gcgaccattg gcaaactgac cctgaaattt atttgcacca
     4081 ccggcaaact accggtgccg tggccgaccc tggtgaccac cttaacctat ggcgtgcagt
     4141 gctttagccg ctatccggat catatgaaac gccatgattt ttttaaaagc gcgatgccgg
     4201 aaggctatgt gcaggaacgc accattagct ttaaagatga tggcaaatat aaaacccgcg
     4261 cggtggtgaa atttgaaggc gataccctgg tgaaccgcat tgaactgaaa ggcaccgatt
     4321 ttaaagaaga tggcaacatt ctggggcata aactggaata taacacagat cagatgccgc
     4381 agatggttac tcttcttcga gttctgagaa ttgttatctt aataagaata tttcgcctgg
     4441 cttcacagaa gaaacaactt gaagtggtaa cctaggtaaa ttcactcctc aggtgcaggc
     4501 tgcctatcag aaggtggtgg ctggtgtggc caatgccctg gctcacaaat accactgaga
     4561 tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc
     4621 tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc
     4681 actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta
     4741 gagtttggca acatatgccc atatgctggc tgccatgaac aaaggttggc tataaagagg
     4801 tcatcagtat atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac
     4861 ttgaggttag atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa
     4921 attttcctta catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat
     4981 agctgtccct cttctcttat gaagatcgga tcgctgcaag aattcaagct tggagactaa
     5041 ctacggtgaa agagacttag aaggagattg agagatactg gagttccgcg ttacataact
     5101 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     5161 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     5221 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     5281 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     5341 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtcgaggt
     5401 gagccccacg ttctgcttca ctctccccat ctcccccccc tccccacccc caattttgta
     5461 tttatttatt ttttaattat tttgtgcagc gatgggggcg gggggggggg ggcgcgcgcc
     5521 aggcggggsg gggggggggg gggggggggg gggggggggg gggggggggg ggggggsrgc
     5581 caawcagrgs ggsgcgcycc gaaagtttcc ttttwtggsg rggsggsggc ggcggcggcc
     5641 ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg cgctgccttc gccccgtgcc
     5701 ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt tactcccaca
     5761 ggtgagcggg cgggacggcc cttctcctcc gggctgtaat tagcgcttgg tttaatgacg
     5821 gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg ctccgggagg gccctttgtg
     5881 cggggggagc ggctcggggg gtgcgtgcgt gtgtgtgtgc gtggggagcg ccgcgtgcgg
     5941 ctccgcgctg cccggcggct gtgagcgctg cgggcgcggc gcggggcttt gtgcgctccg
     6001 cagtgtgcgc gaggggagcg cggccggggg cggtgccccg cggtgcgggg ggggctgcga
     6061 ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg tgagcagggg gtgtgggcgc
     6121 gtcggtcggg ctgcaacccc ccctgcaccc ccctccccga gttgctgagc acggcccggc
     6181 ttcgggtgcg gggctccgta cggggcgtgg cgcggggctc gccgtgccgg gcggggggtg
     6241 gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg gccggggrgg gctcgggggr
     6301 ggggcgcggc ggcccccgga gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg
     6361 ccttttatgg taatcgtgcg agagggcgca gggacttcct ttgtcccaaa tctgtgcgga
     6421 gccgaaatct gggaggcgcc gccgcacccc ctctagcggg cgcggggcga agcggtgcgg
     6481 cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg tcgccgcgcc gccgtcccct
     6541 tctccctctc cagcctcggg gctgtccgcg gggggacggc tgccttcggg ggggacgggg
     6601 cagggcgggg ttcggcttct ggcgtgtgac cggcggctct agagcctctg ctaaccatgt
     6661 tcatgccttc ttctttttcc tacagctcct gggcaacgtg ctggttattg tgctgtctca
     6721 tcattttggc aaagaattcc gatcgcgcca ccatgctgca gaacgagctt gctcttaagt
     6781 tggctggact tgatattaac aagactggag gaggttctca tcatcatcat catcatggta
     6841 tggctagcat gactggtgga cagcaaatgg gtcgggatct gtacgacgat gacgataagg
     6901 atctcgcaac aatggtcgac tcatcgcgac gtaagtggaa taagtggggt cacgcagtca
     6961 gagctatagg tcggctgagc tcagcgaaca acaccgaaat gatgtaccca gcggatggtg
     7021 gtctgcgtgg ttacactcac atggcgctga aagttgatgg cggcggtcac ctgtcctgtt
     7081 ctttcgtgac cacctaccgc tccaaaaaga ctgtcggcaa cattaagatg cctgccattc
     7141 attacgtcag ccaccgtctg gagcgcctgg aggagagcga taacgaaatg tttgtcgtac
     7201 agcgtgaaca cgcagttgcc aagtttgtgg gcctgggtgg tggcggcggt accggaggga
     7261 gcatgaactc cctgatcaag gagaacatgc gtatgaaagt ggttctggaa ggctccgtaa
     7321 acggccacca gttcaaatgc actggtgaag gcgaaggcaa cccgtatatg ggcacccaga
     7381 ctatgcgtat caaagtgatc gagggtggtc cgctgccgtt tgcgttcgac atcctggcga
     7441 cgtcctttat gtatggctcc cgtaccttca tcaaatatcc gaaaggcatc ccggatttct
     7501 ttaagcagtc cttcccggaa ggttttacct gggaacgtgt gacccgttac gaggacggcg
     7561 gcgtaattac cgttatgcaa gacacgtctc tggaggatgg ctgcctggtg tatcacgtgc
     7621 aggttcgcgg tgtgaacttc ccgagcaatg gtgctgtaat gcaaaagaaa accaaaggtt
     7681 gggagcctac ggactcccaa ctgactgaag agcagatcgc agaatttaaa gaggctttct
     7741 ccctatttga caaggacggg gatgggacaa taacaaccaa ggagatgggg acggtgatgc
     7801 ggtccctggg gcagaacccc acagaagcag agctgcagga catgatcaat gaagtagatg
     7861 ccgacggtga cggcacaatc gacttccctg agttcctgat tatgatggca ggcaaaatga
     7921 aatacacaga cagtgaagaa gaaattagag aagcgttcgg cgtgtttgat aaggatggca
     7981 atggctacat cagtgcagca gagcttcgcc acgtgatgac aaaccttgga gagaagttaa
     8041 cagatgaaga ggttgatgaa atgatcaggg aagcagacag cgatggggat ggtcaggtaa
     8101 actacgaaga gtttgtacaa atgatgacag cgaagtaggt aaattcactc ctcaggtgca
     8161 ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca aataccactg
     8221 agatcttttt ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac
     8281 ttctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct
     8341 ctcactcgga aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt
     8401 ttagagtttg gcaacatatg cccatatgct ggctgccatg aacaaaggtt ggctataaag
     8461 aggtcatcag tatatgaaac agccccctgc tgtccattcc ttattccata gaaaagcctt
     8521 gacttgaggt tagatttttt ttatattttg ttttgtgtta tttttttctt taacatccct
     8581 aaaattttcc ttacatgttt tactagccag atttttcctc ctctcctgac tactcccagt
     8641 catagctgtc cctcttctct tatgaagatc ggatcgctac tagaattcga gctcggagac
     8701 taactacggt gaaagagact tagaaggaga ttgagagata ctggagttcc gcgttacata
     8761 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     8821 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     8881 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     8941 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     9001 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtcga
     9061 ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac ccccaatttt
     9121 gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg ggggggsgsg
     9181 cgccrggggg gggggggggg gggggggggg gggggggggg gggggggggg ggggsggsgg
     9241 crgccaawca grgsggsgcg cyccgaaagt ttccttttwt ggcgrggcgg cggsggcggc
     9301 ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg
     9361 tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc gcgttactcc
     9421 cacaggtgag cgggcgggac ggcccttctc ctccgggctg taattagcgc ttggtttaat
     9481 gacggcttgt ttcttttctg tggctgcgtg aaagccttga ggggctccgg gagggccctt
     9541 tgtgcggggg gagcggctcg gggggtgcgt gcgtgtgtgt gtgcgtgggg agcgccgcgt
     9601 gcggctccgc gctgcccggc ggctgtgagc gctgcgggcg cggcgcgggg ctttgtgcgc
     9661 tccgcagtgt gcgcgagggg agcgcggccg ggggcggtgc cccgcggtgc ggggggggct
     9721 gcgaggggaa caaaggctgc gtgcggggtg tgtgcgtggg ggggtgagca gggggtgtgg
     9781 gcgcgtcggt cgggctgcaa cccccccctg cacccccctc cccgagttgc tgagcacggc
     9841 ccggcttcgg gtgcggggct ccgtacgggg cgtggcgcgg ggctcgccgt gccgggcggg
     9901 gggtggcggc aggtgggggt gccgggcggg gcggggccgc ctcgggccgg ggrgggctcg
     9961 ggggaggggc gcggcggccc ccggagcgcc ggcggctgtc gaggcgcggc gagccgcagc
    10021 cattgccttt tatggtaatc gtgcgagagg gcgcagggac ttcctttgtc ccaaatctgt
    10081 gcggagccga aatctgggag gcgccgccgc accccctcta gcgggcgcgg ggcgaagcgg
    10141 tgcggcgccg gcaggaagga aatgggcggg gagggccttc gtgcgtcgcc gcgccgccgt
    10201 ccccttctcc ctctccagcc tcggggctgt ccgcgggggg acggctgcct tcggggggga
    10261 cggggcaggg cggggttcgg cttctggcgt gtgaccggcg gctctagagc ctctgctaac
    10321 catgttcatg ccttcttctt tttcctacag ctcctgggca acgtgctggt tattgtgctg
    10381 tctcatcatt ttggcaaaga attccgatcg cgccaccatg ggctgcgtgt gcagcagcaa
    10441 ccccgagcta ccggtcgcca caatggtagc aggtcatgcc tctggcagcc ccgcattcgg
    10501 gaccgcctct cattcgaatt gcgaacatga agagatccac ctcgccggct cgatccagcc
    10561 gcatggcgcg cttctggtcg tcagcgaaca tgatcatcgc gtcatccagg ccagcgccaa
    10621 cgccgcggaa tttctgaatc tcggaagcgt actcggcgtt ccgctcgccg agatcgacgg
    10681 cgatctgttg atcaagatcc tgccgcatct cgatcccacc gccgaaggca tgccggtcgc
    10741 ggtgcgctgc cggatcggca atccctctac ggagtactgc ggtctgatgc atcggcctcc
    10801 ggaaggcggg ctgatcatcg aactcgaacg tgccggcccg tcgatcgatc tgtcaggcac
    10861 gctggcgccg gcgctggagc ggatccgcac ggcgggttca ctgcgcgcgc tgtgcgatga
    10921 caccgtgctg ctgtttcagc agtgcaccgg ctacgaccgg gtgatggtgt atcgtttcga
    10981 tgagcaaggc cacggcctgg tattctccga gtgccatgtg cctgggctcg aatcctattt
    11041 cggcaaccgc tatccgtcgt cgctggtccc gcagatggcg cggcagctgt acgtgcggca
    11101 gcgcgtccgc gtgctggtcg acgtcaccta tcagccggtg ccgctggagc cgcggctgtc
    11161 gccgctgacc gggcgcgatc tcgacatgtc gggctgcttc ctgcgctcga tgtcgccgat
    11221 ccatctgcag ttcctgaagg acatgggcgt gcgcgccacc ctggcggtgt cgctggtggt
    11281 cggcggcaag ctgtggggcc tggttgtctg tcaccattat ctgccgcgct tcatccgttt
    11341 cgagctgcgg gcgatctgca aacggctcgc cgaaaggatc gcgacgcgga tcaccgcgct
    11401 tgagagctaa gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt
    11461 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
    11521 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
    11581 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
    11641 tcaatgtatc ttaaggcgct ttacgaattc ccatggggag gccgcgaatt ctgaaagacc
    11701 ccacctgtag gtttggcaag cccagggatg tacgtcccta acccgctagg gggcagcaac
    11761 tagtcccagg cctgcactgc cgcctgccgg caggggtcca gtcgctagcg catgcctgca
    11821 ctactggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
    11881 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
    11941 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
    12001 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
    12061 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
    12121 gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc
    12181 ccccccyccc cmcccccaat tttgtwttta tttwtttttt aattwttttg kgcrgsgrkg
    12241 ggggsggggg gggggggggc gcgcgccagg cggggcgggg cggggcgagg ggcggggcgg
    12301 ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa agtttccttt
    12361 tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc gggcgggagt
    12421 cgctgcgcgc tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg
    12481 gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg
    12541 ctgtaattag cgcttggttt aatgacggct tgtttctttt ctgtggctgc gtgaaagcct
    12601 tgaggggctc cgggagggcc ctttgtgcgg ggggagcggc tcggggggtg cgtgcgtgtg
    12661 tgtgtgcgtg gggagcgccg cgtgcggctc cgcgctgccc ggcggctgtg agcgctgcgg
    12721 gcgcggcgcg gggctttgtg cgctccgcag tgtgcgcgag gggagcgcgg ccgggggcgg
    12781 tgccccgcgg tgcggggggg gctgcgaggg gaacaaaggc tgcgtgcggg gtgtgtgcgt
    12841 gggggggtga gcagggggtg tgggcgcgtc ggtcgggctg caaccccccc tgcacccccc
    12901 tccccgagtt gctgagcacg gcccggcttc gggtgcgggg ctccgtacgg ggcgtggcgc
    12961 ggggctcgcc gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg gggcggggcc
    13021 gcctcgggcc ggggrgggct cgggggrggg gcgcggcggc ccccggagcg ccggcggctg
    13081 tcgaggcgcg gcgagccgca gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg
    13141 acttcctttg tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc
    13201 tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct
    13261 tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag cctcggggct gtccgcgggg
    13321 ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc gtgtgaccgg
    13381 cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac agctcctggg
    13441 caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattccgat cgcgccacca
    13501 tggagacgac tgtgaggtat gaacaggggt cagagctcac taaaacttcg agctctccaa
    13561 cagcagatga gcccacgata aagattgatg atggtcgtga tgagggtaat gaacaagaca
    13621 gctgttccaa taccattagg agaaaaattt ccccgtttgt gatgtcattt ggattcagag
    13681 tatttggagt tgtgcttatc attgtagaca tcatagtggt gattgtggat ctggccatca
    13741 gtgagaagaa aagaggcatt agagagattc ttgaaggtgt ttccctggct atagcactct
    13801 tcttccttgt tgatgttctc atgagagtgt ttgttgaagg cttcaagaac tatttccggt
    13861 ccaaactgaa tactttggat gcagtcatag tagtgggcac tctgctaatt aatatgacct
    13921 actccttctc tgaccttagc tttaacagcc ataacgtgta tattaccgcg gataaacaga
    13981 aaaacggcat taaagcgaac tttaccgtgc gccataacgt ggaagatggc agcgtgcagc
    14041 tggcggatca ttatcagcag aacaccccga ttggcgatgg cccggtgctg ctgccggata
    14101 accattatct gagcacccag accgtgctga gcaaagatcc gaacgaaaaa cgcgatcaca
    14161 tggtgctgct ggaatttgtg accgcagcgg gcattacaca cggcatggat gaactgtatg
    14221 gcggcaccgg cggcagcgcg agccagggcg aagaactgtt taccggcgtg gtgccgattc
    14281 tggtggaact ggatggcgat gtgaacggcc ataaatttag cgtgcgcggc gaaggcgaag
    14341 gcgatgcgac cattggcaaa ctgaccctga aatttatttg caccaccggc aaactaccgg
    14401 tgccgtggcc gaccctggtg accaccttaa cctatggcgt gcagtgcttt agccgctatc
    14461 cggatcatat gaaacgccat gattttttta aaagcgcgat gccggaaggc tatgtgcagg
    14521 aacgcaccat tagctttaaa gatgatggca aatataaaac ccgcgcggtg gtgaaatttg
    14581 aaggcgatac cctggtgaac cgcattgaac tgaaaggcac cgattttaaa gaagatggca
    14641 acattctggg gcataaactg gaatataaca cagatcagat gccgcagatg gttactcttc
    14701 ttcgagttct gagaattgtt atcttaataa gaatatttcg cctggcttca cagaagaaac
    14761 aacttgaagt ggtaacctag gtaaattcac tcctcaggtg caggctgcct atcagaaggt
    14821 ggtggctggt gtggccaatg ccctggctca caaataccac tgagatcttt ttccctctgc
    14881 caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa
    14941 atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata
    15001 tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata
    15061 tgcccatatg ctggctgcca tgaacaaagg ttggctataa agaggtcatc agtatatgaa
    15121 acagccccct gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt
    15181 ttttatattt tgttttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt
    15241 tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct
    15301 cttatgaaga tcggatcgct cagagaggat gcacatgtga ccgagggagg agctgcaaga
    15361 actcttcctc acgataactt cgtataggag actttatacg aagttaagcg ctcactggcc
    15421 gtcgttttac agggacacga agtgatccgt ttaaactatc agtgtttgac aggatatatt
    15481 ggcgggtaaa cctaagagaa aagagcgttt attagaataa tcggatattt aaaagggcgt
    15541 gaaaaggttt atccgttcgt ccatttgtat gtgccagccg cctttgcgac gctcaccggg
    15601 ctggttgccc tcgccgctgg gctggcggcc gtctatggcc ctgcaaacgc gccagaaacg
    15661 ccgtcgaagc cgtgtgcgag acaccgcggc cgccggcgtt gtggatacct cgcggaaaac
    15721 ttggccctca ctgacagatg aggggcggac gttgacactt gaggggccga ctcacccggc
    15781 gcggcgttga cagatgaggg gcaggctcga tttcggccgg cgacgtggag ctggccagcc
    15841 tcgcaaatcg gcgaaaacgc ctgattttac gcgagtttcc cacagatgat gtggacaagc
    15901 ctggggataa gtgccctgcg gtattgacac ttgaggggcg cgactactga cagatgaggg
    15961 gcgcgatcct tgacacttga ggggcagagt gctgacagat gaggggcgca cctattgaca
    16021 tttgaggggc tgtccacagg cagaaaatcc agcatttgca agggtttccg cccgtttttc
    16081 ggccaccgct aacctgtctt ttaacctgct tttaaaccaa tatttataaa ccttgttttt
    16141 aaccagggct gcgccctgtg cgcgtgaccg cgcacgccga aggggggtgc ccccccttct
    16201 cgaaccctcc cggcccgcta acgcgggcct cccatccccc caggggctgc gcccctcggc
    16261 cgcgaacggc ctcaccccaa aaatggcagc gctggccaat tcccgagtgc gcggaacccc
    16321 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
    16381 ataaatgctt caataatatt gaaaaaggaa gagtatggct aaaatgagaa tatcaccgga
    16441 attgaaaaaa ctgatcgaaa aataccgctg cgtaaaagat acggaaggaa tgtctcctgc
    16501 taaggtatat aagctggtgg gagaaaatga aaacctatat ttaaaaatga cggacagccg
    16561 gtataaaggg accacctatg atgtggaacg ggaaaaggac atgatgctat ggctggaagg
    16621 aaagctgcct gttccaaagg tcctgcactt tgaacggcat gatggctgga gcaatctgct
    16681 catgagtgag gccgatggcg tcctttgctc ggaagagtat gaagatgaac aaagccctga
    16741 aaagattatc gagctgtatg cggagtgcat caggctcttt cactccatcg acatatcgga
    16801 ttgtccctat acgaatagct tagacagccg cttagccgaa ttggattact tactgaataa
    16861 cgatctggcc gatgtggatt gcgaaaactg ggaagaggac actccattta aagatccgcg
    16921 cgagctgtat gattttttaa agacggaaaa gcccgaagag gaacttgtct tttcccacgg
    16981 cgacctggga gacagcaaca tctttgtgaa agatggcaaa gtaagtggct ttattgatct
    17041 tgggagaagc ggcagggcgg acaagtggta tgacattgcc ttctgcgtcc ggtcgctcag
    17101 ggaggatatc ggggaagaac agtatgtcga gctatttttt gacttactgg ggatcaagcc
    17161 tgattgggag aaaataaaat attatatttt actggatgaa ttgttttagc tgtcagacca
    17221 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
    17281 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
    17341 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
    17401 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
    17461 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
    17521 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
    17581 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
    17641 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
    17701 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
    17761 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
    17821 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
    17881 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
    17941 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
    18001 ggcagatcct agatgtggcg caacgatgcc ggcgacaagc aggagcgcac cgacttcttc
    18061 cgcatcaagt gttttggctc tcaggccgag gcccacggca agtatttggg caaggggtcg
    18121 ctggtattcg tgcagggcaa gattcggaat accaagtacg agaaggacgg ccagacggtc
    18181 tacgggaccg acttcattgc cgataaggtg gattatctgg acaccaaggc accaggcggg
    18241 tcaaatcagg aataagggca cattgccccg gcgtgagtcg gggcaatccc gcaaggaggg
    18301 tgaatgaatc ggacgtttga ccggaaggca tacaggcaag aactgatcga cgcggggttt
    18361 tccgccgagg atgccgaaac catcgcaagc cgcaccgtca tgcgtgcgcc ccgcgaaacc
    18421 ttccagtccg tcggctcgat ggtccagcaa gctacggcca agatcgagcg cgacagcgtg
    18481 caactggctc cccctgccct gcccgcgcca tcggccgccg tggagcgttc gcgtcgtctt
    18541 gaacaggagg cggcaggttt ggcgaagtcg atgaccatcg acacgcgagg aactatgacg
    18601 accaagaagc gaaaaaccgc cggcgaggac ctggcaaaac aggtcagcga ggccaagcag
    18661 gccgcgttgc tgaaacacac gaagcagcag atcaaggaaa tgcagctttc cttgttcgat
    18721 attgcgccgt ggccggacac gatgcgagcg atgccaaacg acacggcccg ctctgccctg
    18781 ttcaccacgc gcaacaagaa aatcccgcgc gaggcgctgc aaaacaaggt cattttccac
    18841 gtcaacaagg acgtgaagat cacctacacc ggcgtcgagc tgcgggccga cgatgacgaa
    18901 ctggtgtggc agcaggtgtt ggagtacgcg aagcgcaccc ctatcggcga gccgatcacc
    18961 ttcacgttct acgagctttg ccaggacctg ggctggtcga tcaatggccg gtattacacg
    19021 aaggccgagg aatgcctgtc gcgcctacag gcgacggcga tgggcttcac gtccgaccgc
    19081 gttgggcacc tggaatcggt gtcgctgctg caccgcttcc gcgtcctgga ccgtggcaag
    19141 aaaacgtccc gttgccaggt cctgatcgac gaggaaatcg tcgtgctgtt tgctggcgac
    19201 cactacacga aattcatatg ggagaagtac cgcaagctgt cgccgacggc ccgacggatg
    19261 ttcgactatt tcagctcgca ccgggagccg tacccgctca agctggaaac cttccgcctc
    19321 atgtgcggat cggattccac ccgcgtgaag aagtggcgcg agcaggtcgg cgaagcctgc
    19381 gaagagttgc gaggcagcgg cctggtggaa cacgcctggg tcaatgatga cctggtgcat
    19441 tgcaaacgct agggccttgt ggggtcagtt ccggctgggg gttcagcagc ccctgctcgg
    19501 atctgttgga ccggacagta gtcatggttg atgggctgcc tgtatcg
//
