LOCUS       sequence-462079-        9941 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bxb1-GT donor plasmid with downstream tandem syntax for all-in-one 
            doxycycline-inducible expression (empty) .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9941)
  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 9941)
  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-GT_AIO-TetOn_Down-Tandem Donor
FEATURES             Location/Qualifiers
     source          1..9941
                     /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     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     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    complement(1802..1835)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    1881..1899
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1917..1935
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1953..1971
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1989..2007
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2025..2043
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2061..2079
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2097..2115
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2160..2184
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2263..2272
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2267..2276
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2280..2301
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2316..2321,2322..2339,2340..2346)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2354..2370
                     /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     2378..2394
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2390..2713
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    2407..2463
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(2464..2480)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2779..3367
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2832..2852)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3250..3261)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     enhancer        3424..3803
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     4973..4995
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     5057..5076
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             5108..5851
                     /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             5852..5905
                     /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             5906..6613
                     /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             6638..6658
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6662..6682
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6686..6706
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(6771..6788)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6777..7001
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    complement(7030..7062)
                     /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(7070..7086)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      7299..7754
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7386..7405)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7596..7617
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7890..7912
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7950..7968)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8036..8140
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8141..8209,8210..9001)
                     /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(8359..8378)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9172..9760
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9661..9680
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9914..9931
                     /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 gcggtctccg tcgtcaggat catccggatc
     1801 cataacttcg tatagcatac attatacgaa gttatcatga tattcggcaa gcaggcatcg
     1861 actagttaat taaggtttac tccctatcag tgatagagaa cgtatgaaga gtttactccc
     1921 tatcagtgat agagaacgta tgcagacttt actccctatc agtgatagag aacgtataag
     1981 gagtttactc cctatcagtg atagagaacg tatgaccagt ttactcccta tcagtgatag
     2041 agaacgtatc tacagtttac tccctatcag tgatagagaa cgtatatcca gtttactccc
     2101 tatcagtgat agagaacgta taagctttag gcgtgtacgg tgggcgccta taaaagcaga
     2161 gctcgtttag tgaaccgtca gatcgcctgg agcaattcca caacactttt gtcttatacc
     2221 aactttccgt accacttcct accctcgtaa acttaagaat tcgccaccat ggcggccgct
     2281 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     2341 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     2401 tacgccaagc ttgcatgcct gcaggtcgac tctagaggat ccccgggtac cgagctcgaa
     2461 ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
     2521 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
     2581 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct
     2641 ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc
     2701 tgatgccgca tagcccagcc cgcctaatga gcgggctttt ttttgaacaa attaagccta
     2761 taacgcggaa gcggccgcaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     2821 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     2881 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     2941 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     3001 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     3061 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     3121 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg
     3181 tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     3241 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     3301 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     3361 tccccgccct ttgggccgcc tccccgcgct agtgctgggg atgcggtggg ctctatgggt
     3421 ctcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
     3481 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     3541 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     3601 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     3661 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     3721 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     3781 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
     3841 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
     3901 cgatgggggc gggggggggg ggggcgcgcg ccaggcgggg cggggggggg gggggggggg
     3961 gggggggggg gggggggggg gggggggggg gcccaggggg gggggggggg gggggaggtt
     4021 tctttttttg gggggggggg gggggggggg gcccttaaaa aaaggaaggg gggggggggg
     4081 gggggggggc ggggggggct tccccccccg cccccccccc cccccccccc cccccccccc
     4141 ccccgccgcc gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct
     4201 ccgggctgta attagcgctt ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa
     4261 agccttgagg ggctccggga gggccctttg tgcgggggga gcggctcggg gggtgcgtgc
     4321 gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc
     4381 tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg
     4441 ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg
     4501 tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac
     4561 ccccctcccc gagttgctga gcgcggcccg gcttcgggtg cggggctccg tacggggcgt
     4621 ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggcccc
     4681 cggagcgccg gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg
     4741 tgcgagaggg cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg
     4801 cgccgccgca ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa
     4861 atgggcgggg agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct
     4921 cggggctgtc cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc
     4981 ttctggcgtg tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt
     5041 ttcctacagc tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa
     5101 tgtcgacatg agccggctgg acaagagcaa agtgatcaac agcgccctgg aactgctgaa
     5161 cggcgtgggc atcgagggcc tgaccacccg gaagctggcc cagaaactgg gcgtggaaca
     5221 gcccaccctg tactggcacg tgaagaacaa gcgggccctg ctggacgccc tgcccatcga
     5281 gatgctggac cggcaccaca cccacagctg ccctctggaa ggcgagagct ggcaggactt
     5341 cctgcggaac aacgccaaga gctacagatg cgccctgctg agccaccggg acggcgccaa
     5401 agtgcacctg ggcaccagac ccaccgagaa gcagtacgag acactggaaa accagctggc
     5461 cttcctgtgc cagcagggct tcagcctgga aaacgccctg tacgccctga gcgccgtggg
     5521 ccacttcacc ctgggctgcg tgctggaaga acaggaacac caggtcgcca aagaggaaag
     5581 agagacaccc accaccgaca gcatgccccc cctgctgaag caggccatcg agctgttcga
     5641 cagacagggc gccgagcccg ccttcctgtt cggcctggaa ctgatcatct gcggcctcga
     5701 gaagcagctg aagtgcgaga gcggcggacc caccgacgcc ctggacgact tcgacctgga
     5761 catgctgccc gccgatgccc tggatgattt tgatctggat atgctccctg ctgacgctct
     5821 cgatgacttc gatctcgata tgctgccagg cgagggcagg ggaagtcttc taacatgcgg
     5881 ggacgtggag gaaaatcccg gccccgtgtc taagggcgaa gagctgatta aggagaacat
     5941 gcacatgaag ctgtacatgg agggcaccgt ggacaaccat cacttcaagt gcacatccga
     6001 gggcgaaggc aagccctacg agggcaccca gaccatgaga atcaaggtgg tcgagggcgg
     6061 ccctctcccc ttcgccttcg acatcctggc tactagcttc ctctacggca gcaagacctt
     6121 catcaaccac acccagggca tccccgactt cttcaagcag tccttccctg agggcttcac
     6181 atgggagaga gtcaccacat acgaagacgg gggcgtgctg accgctaccc aggacaccag
     6241 cctccaggac ggctgcctca tctacaacgt caagatcaga ggggtgaact tcacatccaa
     6301 cggccctgtg atgcagaaga aaacactcgg ctgggaggcc ttcaccgaga cgctgtaccc
     6361 cgctgacggc ggcctggaag gcagaaacga catggccctg aagctcgtgg gcgggagcca
     6421 tctgatcgca aacgccaaga ccacatatag atccaagaaa cccgctaaga acctcaagat
     6481 gcctggcgtc tactatgtgg actacagact ggaaagaatc aaggaggcca acaacgagac
     6541 ctacgtcgag cagcacgagg tggcagtggc cagatactgc gacctcccta gcaaactggg
     6601 gcacaagctt aattccggac tcagatctcg agctgatcca aaaaagaaga gaaaggtaga
     6661 tccaaaaaag aagagaaagg tagatccaaa aaagaagaga aaggtaggat ccaccggatc
     6721 tagataactg atcataatca gccataccac atttgaaacc cgctgatcag cctcgactgt
     6781 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     6841 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     6901 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     6961 agacaatagc aggcatgctg gggatgcggt gggctctatg gctagccgct gcaagaactc
     7021 ttcctcacga taacttcgta taggagactt tatacgaagt taagcgctca ctggccgtcg
     7081 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     7141 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     7201 agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt
     7261 gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt
     7321 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc
     7381 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     7441 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     7501 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     7561 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     7621 aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt
     7681 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     7741 aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     7801 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     7861 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     7921 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     7981 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     8041 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     8101 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     8161 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     8221 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     8281 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     8341 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     8401 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     8461 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     8521 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     8581 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     8641 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     8701 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     8761 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     8821 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     8881 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     8941 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     9001 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     9061 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     9121 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     9181 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     9241 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     9301 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     9361 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     9421 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     9481 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     9541 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     9601 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     9661 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     9721 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     9781 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     9841 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     9901 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa g
//
