LOCUS       sequence-461788-       10200 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bxb1-GA donor plasmid with upstream tandem syntax for all-in-one 
            doxycycline-inducible mScarlet expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10200)
  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 10200)
  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_AIO-TetOn_mScarlet_Up-Tandem
FEATURES             Location/Qualifiers
     source          1..10200
                     /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        1897..2276
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2278..2552
                     /label=chicken beta-actin promoter
     primer_bind     3445..3467
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3529..3548
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             3580..4323
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     CDS             4324..4377
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             4378..5085
                     /codon_start=1
                     /product="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMR
                     IKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGV
                     LTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDM
                     ALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVA
                     RYCDLPSKLGHKLN"
     CDS             5110..5130
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5134..5154
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5158..5178
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(5243..5260)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5249..5473
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    5498..5516
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5534..5552
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5570..5588
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5606..5624
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5642..5660
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5678..5696
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5714..5732
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     5777..5801
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      5880..5889
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5886..6581
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(6019..6037)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     6287..6306
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    6585..7173
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6638..6658)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7056..7067)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    7258..7291
                     /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(7299..7315)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7329..7345)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7338..7360)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7558..8013
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7645..7664)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7855..7876
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8030..8049)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8149..8171
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8209..8227)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8295..8399
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8400..8468,8469..9260)
                     /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(8618..8637)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9431..10019
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9920..9939
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10173..10190
                     /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 ctagttaatt aagctcgaca ttgattattg actagttatt
     1921 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
     1981 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     2041 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     2101 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
     2161 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     2221 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtcg
     2281 aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt
     2341 tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg ggggggcgcg
     2401 cgccgggggg ggcggggcgg ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg
     2461 cagccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc
     2521 ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg
     2581 tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc gcgttactcc
     2641 cacaggtgag cgggcgggac ggcccttctc ctccgggctg taattagcgc ttggtttaat
     2701 gacggcttgt ttcttttctg tggctgcgtg aaagccttga ggggctccgg gagggccctt
     2761 tgtgcggggg gagcggctcg gggggtgcgt gcgtgtgtgt gtgcgtgggg agcgccgcgt
     2821 gcggctccgc gctgcccggc ggctgtgagc gctgcgggcg cggcgcgggg ctttgtgcgc
     2881 tccgcagtgt gcgcgagggg agcgcggccg ggggcggtgc cccgcggtgc ggggggggct
     2941 gcgaggggaa caaaggctgc gtgcggggtg tgtgcgtggg ggggtgagca gggggtgtgg
     3001 gcgcgtcggt cgggctgcaa ccccccctgc acccccctcc ccgagttgct gagcgcggcc
     3061 cggcttcggg tgcggggctc cgtacggggc gtggcgcggg gctcgccgtg ccgggcgggg
     3121 ggtggcggca ggtgggggtg ccgggcggcc cccggagcgc cggcggctgt cgaggcgcgg
     3181 cgagccgcag ccattgcctt ttatggtaat cgtgcgagag ggcgcaggga cttcctttgt
     3241 cccaaatctg tgcggagccg aaatctggga ggcgccgccg caccccctct agcgggcgcg
     3301 gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt cgtgcgtcgc
     3361 cgcgccgccg tccccttctc cctctccagc ctcggggctg tccgcggggg gacggctgcc
     3421 ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc ggctctagag
     3481 cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc aacgtgctgg
     3541 ttattgtgct gtctcatcat tttggcaaag aatgtcgaca tgagccggct ggacaagagc
     3601 aaagtgatca acagcgccct ggaactgctg aacggcgtgg gcatcgaggg cctgaccacc
     3661 cggaagctgg cccagaaact gggcgtggaa cagcccaccc tgtactggca cgtgaagaac
     3721 aagcgggccc tgctggacgc cctgcccatc gagatgctgg accggcacca cacccacagc
     3781 tgccctctgg aaggcgagag ctggcaggac ttcctgcgga acaacgccaa gagctacaga
     3841 tgcgccctgc tgagccaccg ggacggcgcc aaagtgcacc tgggcaccag acccaccgag
     3901 aagcagtacg agacactgga aaaccagctg gccttcctgt gccagcaggg cttcagcctg
     3961 gaaaacgccc tgtacgccct gagcgccgtg ggccacttca ccctgggctg cgtgctggaa
     4021 gaacaggaac accaggtcgc caaagaggaa agagagacac ccaccaccga cagcatgccc
     4081 cccctgctga agcaggccat cgagctgttc gacagacagg gcgccgagcc cgccttcctg
     4141 ttcggcctgg aactgatcat ctgcggcctc gagaagcagc tgaagtgcga gagcggcgga
     4201 cccaccgacg ccctggacga cttcgacctg gacatgctgc ccgccgatgc cctggatgat
     4261 tttgatctgg atatgctccc tgctgacgct ctcgatgact tcgatctcga tatgctgcca
     4321 ggcgagggca ggggaagtct tctaacatgc ggggacgtgg aggaaaatcc cggccccgtg
     4381 tctaagggcg aagagctgat taaggagaac atgcacatga agctgtacat ggagggcacc
     4441 gtggacaacc atcacttcaa gtgcacatcc gagggcgaag gcaagcccta cgagggcacc
     4501 cagaccatga gaatcaaggt ggtcgagggc ggccctctcc ccttcgcctt cgacatcctg
     4561 gctactagct tcctctacgg cagcaagacc ttcatcaacc acacccaggg catccccgac
     4621 ttcttcaagc agtccttccc tgagggcttc acatgggaga gagtcaccac atacgaagac
     4681 gggggcgtgc tgaccgctac ccaggacacc agcctccagg acggctgcct catctacaac
     4741 gtcaagatca gaggggtgaa cttcacatcc aacggccctg tgatgcagaa gaaaacactc
     4801 ggctgggagg ccttcaccga gacgctgtac cccgctgacg gcggcctgga aggcagaaac
     4861 gacatggccc tgaagctcgt gggcgggagc catctgatcg caaacgccaa gaccacatat
     4921 agatccaaga aacccgctaa gaacctcaag atgcctggcg tctactatgt ggactacaga
     4981 ctggaaagaa tcaaggaggc caacaacgag acctacgtcg agcagcacga ggtggcagtg
     5041 gccagatact gcgacctccc tagcaaactg gggcacaagc ttaattccgg actcagatct
     5101 cgagctgatc caaaaaagaa gagaaaggta gatccaaaaa agaagagaaa ggtagatcca
     5161 aaaaagaaga gaaaggtagg atccaccgga tctagataac tgatcataat cagccatacc
     5221 acatttgaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     5281 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     5341 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     5401 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     5461 gtgggctcta tggatcgact agttaattaa ggtttactcc ctatcagtga tagagaacgt
     5521 atgaagagtt tactccctat cagtgataga gaacgtatgc agactttact ccctatcagt
     5581 gatagagaac gtataaggag tttactccct atcagtgata gagaacgtat gaccagttta
     5641 ctccctatca gtgatagaga acgtatctac agtttactcc ctatcagtga tagagaacgt
     5701 atatccagtt tactccctat cagtgataga gaacgtataa gctttaggcg tgtacggtgg
     5761 gcgcctataa aagcagagct cgtttagtga accgtcagat cgcctggagc aattccacaa
     5821 cacttttgtc ttataccaac tttccgtacc acttcctacc ctcgtaaact taagaattcg
     5881 ccaccatggt gagcaagggc gaggcagtga tcaaggagtt catgcggttc aaggtgcaca
     5941 tggagggctc catgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
     6001 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttctcct
     6061 gggacatcct gtcccctcag ttcatgtacg gctccagggc cttcaccaag caccccgccg
     6121 acatccccga ctactataag cagtccttcc ccgagggctt caagtgggag cgcgtgatga
     6181 acttcgagga cggcggcgcc gtgaccgtga cccaggacac ctccctggag gacggcaccc
     6241 tgatctacaa ggtgaagctc cgcggcacca acttccctcc tgacggcccc gtaatgcaga
     6301 agaagacaat gggctgggaa gcgtccaccg agcggttgta ccccgaggac ggcgtgctga
     6361 agggcgacat taagatggcc ctgcgcctga aggacggcgg ccgctacctg gcggacttca
     6421 agaccaccta caaggccaag aagcccgtgc agatgcccgg cgcctacaac gtcgaccgca
     6481 agttggacat cacctcccac aacgaggact acaccgtggt ggaacagtac gaacgctccg
     6541 agggccgcca ctccaccggc ggcatggacg agctgtacaa gtaaaatcaa cctctggatt
     6601 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     6661 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     6721 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     6781 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     6841 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     6901 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     6961 ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct
     7021 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     7081 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     7141 cgagtcggat ctccctttgg gccgcctccc cgccctttgg gccgcctccc cgcgctagtg
     7201 ctggggatgc ggtgccgctg caagaactct tcctcacgga tccgaagttc ctattccgaa
     7261 gttcctattc ttcaaatagt ataggaactt cagcgctcac tggccgtcgt tttacaaagg
     7321 gcgaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     7381 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     7441 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat
     7501 tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg
     7561 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     7621 cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     7681 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     7741 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     7801 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     7861 tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt gatttataag
     7921 ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     7981 cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct
     8041 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     8101 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     8161 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     8221 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     8281 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     8341 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     8401 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     8461 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     8521 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     8581 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     8641 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     8701 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     8761 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     8821 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     8881 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     8941 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     9001 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     9061 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     9121 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     9181 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     9241 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     9301 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     9361 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     9421 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     9481 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     9541 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     9601 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     9661 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     9721 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     9781 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     9841 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     9901 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     9961 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
    10021 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
    10081 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
    10141 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
//
