LOCUS       sequence-462083-        9070 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bxb1-GA donor plasmid for integrating a transcription unit for 
            constitutive expression of a mScarlet-I F-actin reporter (3 copies).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9070)
  AUTHORS   Blanch-Asensio A, Ploessl DS, Wang NB, Mummery CL, Galloway KE, 
            Davis RP
  TITLE     STRAIGHT-IN Dual: a platform for dual, single-copy integrations of 
            DNA payloads and gene circuits into human induced pluripotent stem 
            cells.
  JOURNAL   bioRxiv [Preprint]. 2024 Oct 17:2024.10.17.616637. doi: 
            10.1101/2024.10.17.616637.
  PUBMED    39464154
REFERENCE   2  (bases 1 to 9070)
  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: "bioRxiv 
            [Preprint]. 2024 Oct 17:2024.10.17.616637. doi: 
            10.1101/2024.10.17.616637."
COMMENT     Sequence Label: Bxb1-GA_CAG-ITPKA-3x-mScarlet-I
FEATURES             Location/Qualifiers
     source          1..9070
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        253..463
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        275..471
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      322..457
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        558..1736
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          789..1727
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1684..1704
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1744..1753
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    1756..1793
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attB site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     protein_bind    1808..1855
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     enhancer        1883..2186
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2188..2463
                     /label=chicken beta-actin promoter
     intron          2464..3480
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3404..3426
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3488..3507
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      3542..3551
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3718..3727
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3724..4416
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELY"
     CDS             4435..5127
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELY"
     CDS             5146..5841
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     polyA_signal    5967..6048
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6004..6023)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6058..6077
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    6128..6161
                     /label=FRT3
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision with other FRT3 sites
                     (Bosch et al., 2015)"
     primer_bind     complement(6169..6185)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6199..6215)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6208..6230)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6428..6883
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6515..6534)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6725..6746
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6900..6919)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7019..7041
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7079..7097)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7165..7269
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7270..7338,7339..8130)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7488..7507)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8301..8889
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8790..8809
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9043..9060
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctta
      241 attccctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
      301 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
      361 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
      421 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctttgc
      481 aaagatggat aaagttttaa acagagagga atctttgcag ctaatggacc ttctaggtct
      541 tgaaaggagt gggaattggc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag
      601 tccccgagaa gttgggggga ggggtcggca attgaaccgg tgcctagaga aggtggcgcg
      661 gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag
      721 aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca
      781 gaacacaggt aagtgccgtg tgtggttccc gcgggcctgg cctctttacg ggttatggcc
      841 cttgcgtgcc ttgaattact tccacctggc tgcagtacgt gattcttgat cccgagcttc
      901 gggttggaag tgggtgggag agttcgaggc cttgcgctta aggagcccct tcgcctcgtg
      961 cttgagttga ggcctggcct gggcgctggg gccgccgcgt gcgaatctgg tggcaccttc
     1021 gcgcctgtct cgctgctttc gataagtctc tagccattta aaatttttga tgacctgctg
     1081 cgacgctttt tttctggcaa gatagtcttg taaatgcggg ccaagatctg cacactggta
     1141 tttcggtttt tggggccgcg ggcggcgacg gggcccgtgc gtcccagcgc acatgttcgg
     1201 cgaggcgggg cctgcgagcg cggccaccga gaatcggacg ggggtagtct caagctggcc
     1261 ggcctgctct ggtgcctggc ctcgcgccgc cgtgtatcgc cccgccctgg gcggcaaggc
     1321 tggcccggtc ggcaccagtt gcgtgagcgg aaagatggcc gcttcccggc cctgctgcag
     1381 ggagctcaaa atggaggacg cggcgctcgg gagagcgggc gggtgagtca cccacacaaa
     1441 ggaaaagggc ctttccgtcc tcagccgtcg cttcatgtga ctccacggag taccgggcgc
     1501 cgtccaggca cctcgattag ttctcgagct tttggagtac gtcgtcttta ggttgggggg
     1561 aggggtttta tgcgatggag tttccccaca ctgagtgggt ggagactgaa gttaggccag
     1621 cttggcactt gatgtaattc tccttggaat ttgccctttt tgagtttgga tcttggttca
     1681 ttctcaagcc tcagacagtg gttcaaagtt tttttcttcc atttcaggtg tcgtgaggaa
     1741 ttcgccacca tggccggctt gtcgacgacg gcggactccg tcgtcaggat catccggatc
     1801 cggatccgaa gttcctattc cgaagttcct attctctaga aagtatagga acttcatgat
     1861 attcggcaag caggcatcga cgcgttacat aacttacggt aaatggcccg cctggctgac
     1921 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     1981 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     2041 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     2101 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     2161 acgtattagt catcgctatt accatggtcg aggtgagccc cacgttctgc ttcactctcc
     2221 ccatctcccc cccctcccca cccccaattt tgtatttatt tattttttaa ttattttgtg
     2281 cagcgatggg ggcggggggg gggggggcgc gcgccaggcg gggcggggcg gggcgagggg
     2341 cggggcgggg cgaggcggag aggtgcggcg gcagccaatc agagcggcgc gctccgaaag
     2401 tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg
     2461 gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc
     2521 ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga cggcccttct
     2581 cctccgggct gtaattagcg cttggtttaa tgacggcttg tttcttttct gtggctgcgt
     2641 gaaagccttg aggggctccg ggagggccct ttgtgcgggg ggagcggctc ggggggtgcg
     2701 tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggctccg cgctgcccgg cggctgtgag
     2761 cgctgcgggc gcggcgcggg gctttgtgcg ctccgcagtg tgcgcgaggg gagcgcggcc
     2821 gggggcggtg ccccgcggtg cggggggggc tgcgagggga acaaaggctg cgtgcggggt
     2881 gtgtgcgtgg gggggtgagc agggggtgtg ggcgcgtcgg tcgggctgca accccccctg
     2941 cacccccctc cccgagttgc tgagcacggc ccggcttcgg gtgcggggct ccgtacgggg
     3001 cgtggcgcgg ggctcgccgt gccgggcggg gggtggcggc aggtgggggt gccgggcggg
     3061 gcggggccgc ctcgggccgg ggagggctcg ggggaggggc gcggcggccc ccggagcgcc
     3121 ggcggctgtc gaggcgcggc gagccgcagc cattgccttt tatggtaatc gtgcgagagg
     3181 gcgcagggac ttcctttgtc ccaaatctgt gcggagccga aatctgggag gcgccgccgc
     3241 accccctcta gcgggcgcgg ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg
     3301 gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc ctctccagcc tcggggctgt
     3361 ccgcgggggg acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt
     3421 gtgaccggcg gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag
     3481 ctcctgggca acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attccgatcg
     3541 cgccaccatg gagagctagc gctaccggac tcagatctcg agctcaagct tatgggcatg
     3601 gcgcgaccac ggggcgcggg gccctgcagc cccgggttgg agcgggctcc gcgccggagc
     3661 gtcggggagc tgcgcctgct cttcgaagcg ggcggatctg gatcggatcc accggtcgcc
     3721 accatggtga gcaagggcga ggcagtgatc aaggagttca tgcggttcaa ggtgcacatg
     3781 gagggctcca tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac
     3841 gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttctcctgg
     3901 gacatcctgt cccctcagtt catgtacggc tccagggcct tcatcaagca ccccgccgac
     3961 atccccgact actataagca gtccttcccc gagggcttca agtgggagcg cgtgatgaac
     4021 ttcgaggacg gcggcgccgt gaccgtgacc caggacacct ccctggagga cggcaccctg
     4081 atctacaagg tgaagctccg cggcaccaac ttccctcctg acggccccgt aatgcagaag
     4141 aagacaatgg gctgggaagc gtccaccgag cggttgtacc ccgaggacgg cgtgctgaag
     4201 ggcgacatta agatggccct gcgcctgaag gacggcggcc gctacctggc ggacttcaag
     4261 accacctaca aggccaagaa gcccgtgcag atgcccggcg cctacaacgt cgaccgcaag
     4321 ttggacatca cctcccacaa cgaggactac accgtggtgg aacagtacga acgctccgag
     4381 ggccgccact ccaccggcgg catggacgag ctgtaccagg gtggcagcgg tggcatggtg
     4441 agcaagggcg aggcagtgat caaggagttc atgcggttca aggtgcacat ggagggctcc
     4501 atgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc
     4561 cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttctcctg ggacatcctg
     4621 tcccctcagt tcatgtacgg ctccagggcc ttcatcaagc accccgccga catccccgac
     4681 tactataagc agtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac
     4741 ggcggcgccg tgaccgtgac ccaggacacc tccctggagg acggcaccct gatctacaag
     4801 gtgaagctcc gcggcaccaa cttccctcct gacggccccg taatgcagaa gaagacaatg
     4861 ggctgggaag cgtccaccga gcggttgtac cccgaggacg gcgtgctgaa gggcgacatt
     4921 aagatggccc tgcgcctgaa ggacggcggc cgctacctgg cggacttcaa gaccacctac
     4981 aaggccaaga agcccgtgca gatgcccggc gcctacaacg tcgaccgcaa gttggacatc
     5041 acctcccaca acgaggacta caccgtggtg gaacagtacg aacgctccga gggccgccac
     5101 tccaccggcg gcatggacga gctgtaccag ggtggcagcg gtggcatggt gagcaagggc
     5161 gaggcagtga tcaaggagtt catgcggttc aaggtgcaca tggagggctc catgaacggc
     5221 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc
     5281 aagctgaagg tgaccaaggg tggccccctg cccttctcct gggacatcct gtcccctcag
     5341 ttcatgtacg gctccagggc cttcatcaag caccccgccg acatccccga ctactataag
     5401 cagtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgcc
     5461 gtgaccgtga cccaggatac ctccctggag gacggcaccc tgatctacaa ggtgaagctc
     5521 cgcggcacca acttccctcc tgacggcccc gtaatgcaga agaagacaat gggctgggaa
     5581 gcgtccaccg agcggttgta ccccgaggac ggcgtgctga agggcgacat taagatggcc
     5641 ctgcgcctga aggacggcgg ccgctacctg gcggacttca agaccaccta caaggccaag
     5701 aagcccgtgc agatgcccgg cgcctacaac gtcgaccgca agttggacat cacctcccac
     5761 aacgaggact acaccgtggt ggaacagtac gaacgctccg agggccgcca ctccaccggc
     5821 ggcatggacg agctgtacaa gtaaagcggc cgcgactcta gatcataatc agccatacca
     5881 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     5941 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     6001 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     6061 gtttgtccaa actcatcaat gtatccgctg caagaactct tcctcacgga tccgaagttc
     6121 ctattccgaa gttcctattc ttcaaatagt ataggaactt cagcgctcac tggccgtcgt
     6181 tttacaaagg gcgaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     6241 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     6301 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct
     6361 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa
     6421 ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     6481 gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt
     6541 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     6601 cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt
     6661 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     6721 aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt
     6781 gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa
     6841 aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt
     6901 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     6961 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     7021 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     7081 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     7141 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     7201 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     7261 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     7321 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     7381 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     7441 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     7501 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     7561 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     7621 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     7681 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     7741 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     7801 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     7861 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     7921 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     7981 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     8041 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     8101 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     8161 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     8221 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     8281 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     8341 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     8401 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     8461 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     8521 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     8581 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     8641 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     8701 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     8761 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     8821 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     8881 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     8941 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     9001 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     9061 gaagcggaag
//
