LOCUS       sequence-462080-       10034 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bxb1-GA donor plasmid with divergent 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 10034)
  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 10034)
  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_Divergent Donor
FEATURES             Location/Qualifiers
     source          1..10034
                     /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    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)."
     misc_feature    1952..2540
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             2058..2069
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     2467..2487
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2606..2929)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     primer_bind     2839..2855
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2856..2912
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(2925..2941)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2949..2965
                     /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)."
     promoter        complement(join(2973..2979,2980..2997,2998..3003))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3018..3039
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     regulatory      3043..3052
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3047..3056
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3135..3159)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    3204..3222
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3240..3258
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3276..3294
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3312..3330
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3348..3366
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3384..3402
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3420..3438
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     enhancer        3469..3848
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3850..4123
                     /label=chicken beta-actin promoter
     primer_bind     5016..5038
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     5100..5119
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             5151..5894
                     /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             5895..5948
                     /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             5949..6656
                     /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             6681..6701
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6705..6725
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6729..6749
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(6814..6831)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6820..7044
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    7092..7125
                     /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(7133..7149)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7163..7179)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      7392..7847
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7479..7498)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7689..7710
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7983..8005
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8043..8061)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8129..8233
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8234..8302,8303..9094)
                     /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(8452..8471)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9265..9853
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9754..9773
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10007..10024
                     /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 ctagttaata atgtcgagac ccatagagcc caccgcatcc
     1921 ccagcactag cgcggggagg cggcccaaag ggcggggagg cggcccaaag ggagatccga
     1981 ctcgtctgag ggcgaaggcg aagacgcgga agaggccgca gagccggcag caggccgcgg
     2041 gaaggaaggt ccgctggatt gagggccgaa gggacgtagc agaaggacgt cccgcgcaga
     2101 atccaggtgg caacacaggc gagcagccat ggaaaggacg tcagcttccc cgacaacacc
     2161 acggaattgt cagtgcccaa cagccgagcc cctgtccagc agcgggcaag gcaggcggcg
     2221 atgagttccg ccgtggcaat agggaggggg aaagcgaaag tcccggaaag gagctgacag
     2281 gtggtggcaa tgccccaacc agtgggggtt gcgtcagcaa acacagtgca caccacgcca
     2341 cgttgcctga caacgggcca caactcctca taaagagaca gcaaccagga tttatacaag
     2401 gaggagaaaa tgaaagccat acgggaagca atagcatgat acaaaggcat taaagcagcg
     2461 tatccacata gcgtaaaagg agcaacatag ttaagaatac cagtcaatct ttcacaaatt
     2521 ttgtaatcca gaggttgatt gcggccgctt ccgcgttata ggcttaattt gttcaaaaaa
     2581 aagcccgctc attaggcggg ctgggctatg cggcatcaga gcagattgta ctgagagtgc
     2641 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
     2701 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     2761 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     2821 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acccggggat
     2881 cctctagagt cgacctgcag gcatgcaagc ttggcgtaat catggtcata gctgtttcct
     2941 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     3001 aaagcctggg gtgcctaatg agtgagctaa ctcacattag cggccgccat ggtggcgaat
     3061 tcttaagttt acgagggtag gaagtggtac ggaaagttgg tataagacaa aagtgttgtg
     3121 gaattgctcc aggcgatctg acggttcact aaacgagctc tgcttttata ggcgcccacc
     3181 gtacacgcct aaagcttata cgttctctat cactgatagg gagtaaactg gatatacgtt
     3241 ctctatcact gatagggagt aaactgtaga tacgttctct atcactgata gggagtaaac
     3301 tggtcatacg ttctctatca ctgataggga gtaaactcct tatacgttct ctatcactga
     3361 tagggagtaa agtctgcata cgttctctat cactgatagg gagtaaactc ttcatacgtt
     3421 ctctatcact gatagggagt aaaccttaat taactagtca ttaagctcga cattgattat
     3481 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
     3541 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     3601 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     3661 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     3721 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     3781 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     3841 ttaccatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc cccccctccc
     3901 cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg ggggcggggg
     3961 gggggggcgc gcgccaggcg gggcggggcg gggcgagggg cggggcgggg cgaggcggag
     4021 aggtgcggcg gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg
     4081 gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgcgctg
     4141 ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac
     4201 cgcgttactc ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagcg
     4261 cttggtttaa tgacggcttg tttcttttct gtggctgcgt gaaagccttg aggggctccg
     4321 ggagggccct ttgtgcgggg ggagcggctc ggggggtgcg tgcgtgtgtg tgtgcgtggg
     4381 gagcgccgcg tgcggctccg cgctgcccgg cggctgtgag cgctgcgggc gcggcgcggg
     4441 gctttgtgcg ctccgcagtg tgcgcgaggg gagcgcggcc gggggcggtg ccccgcggtg
     4501 cggggggggc tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg gggggtgagc
     4561 agggggtgtg ggcgcgtcgg tcgggctgca accccccctg cacccccctc cccgagttgc
     4621 tgagcgcggc ccggcttcgg gtgcggggct ccgtacgggg cgtggcgcgg ggctcgccgt
     4681 gccgggcggg gggtggcggc aggtgggggt gccgggcggc ccccggagcg ccggcggctg
     4741 tcgaggcgcg gcgagccgca gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg
     4801 acttcctttg tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc
     4861 tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct
     4921 tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag cctcggggct gtccgcgggg
     4981 ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc gtgtgaccgg
     5041 cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac agctcctggg
     5101 caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaatgtcgac atgagccggc
     5161 tggacaagag caaagtgatc aacagcgccc tggaactgct gaacggcgtg ggcatcgagg
     5221 gcctgaccac ccggaagctg gcccagaaac tgggcgtgga acagcccacc ctgtactggc
     5281 acgtgaagaa caagcgggcc ctgctggacg ccctgcccat cgagatgctg gaccggcacc
     5341 acacccacag ctgccctctg gaaggcgaga gctggcagga cttcctgcgg aacaacgcca
     5401 agagctacag atgcgccctg ctgagccacc gggacggcgc caaagtgcac ctgggcacca
     5461 gacccaccga gaagcagtac gagacactgg aaaaccagct ggccttcctg tgccagcagg
     5521 gcttcagcct ggaaaacgcc ctgtacgccc tgagcgccgt gggccacttc accctgggct
     5581 gcgtgctgga agaacaggaa caccaggtcg ccaaagagga aagagagaca cccaccaccg
     5641 acagcatgcc ccccctgctg aagcaggcca tcgagctgtt cgacagacag ggcgccgagc
     5701 ccgccttcct gttcggcctg gaactgatca tctgcggcct cgagaagcag ctgaagtgcg
     5761 agagcggcgg acccaccgac gccctggacg acttcgacct ggacatgctg cccgccgatg
     5821 ccctggatga ttttgatctg gatatgctcc ctgctgacgc tctcgatgac ttcgatctcg
     5881 atatgctgcc aggcgagggc aggggaagtc ttctaacatg cggggacgtg gaggaaaatc
     5941 ccggccccgt gtctaagggc gaagagctga ttaaggagaa catgcacatg aagctgtaca
     6001 tggagggcac cgtggacaac catcacttca agtgcacatc cgagggcgaa ggcaagccct
     6061 acgagggcac ccagaccatg agaatcaagg tggtcgaggg cggccctctc cccttcgcct
     6121 tcgacatcct ggctactagc ttcctctacg gcagcaagac cttcatcaac cacacccagg
     6181 gcatccccga cttcttcaag cagtccttcc ctgagggctt cacatgggag agagtcacca
     6241 catacgaaga cgggggcgtg ctgaccgcta cccaggacac cagcctccag gacggctgcc
     6301 tcatctacaa cgtcaagatc agaggggtga acttcacatc caacggccct gtgatgcaga
     6361 agaaaacact cggctgggag gccttcaccg agacgctgta ccccgctgac ggcggcctgg
     6421 aaggcagaaa cgacatggcc ctgaagctcg tgggcgggag ccatctgatc gcaaacgcca
     6481 agaccacata tagatccaag aaacccgcta agaacctcaa gatgcctggc gtctactatg
     6541 tggactacag actggaaaga atcaaggagg ccaacaacga gacctacgtc gagcagcacg
     6601 aggtggcagt ggccagatac tgcgacctcc ctagcaaact ggggcacaag cttaattccg
     6661 gactcagatc tcgagctgat ccaaaaaaga agagaaaggt agatccaaaa aagaagagaa
     6721 aggtagatcc aaaaaagaag agaaaggtag gatccaccgg atctagataa ctgatcataa
     6781 tcagccatac cacatttgaa acccgctgat cagcctcgac tgtgccttct agttgccagc
     6841 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     6901 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     6961 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     7021 ctggggatgc ggtgggctct atggctagcc gctgcaagaa ctcttcctca cggatccgaa
     7081 gttcctattc cgaagttcct attcttcaaa tagtatagga acttcagcgc tcactggccg
     7141 tcgttttaca aagggcgaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     7201 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     7261 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc
     7321 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa
     7381 gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     7441 cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc
     7501 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     7561 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
     7621 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     7681 ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc
     7741 ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta
     7801 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct
     7861 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     7921 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     7981 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     8041 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     8101 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     8161 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     8221 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     8281 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     8341 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     8401 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     8461 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     8521 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     8581 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     8641 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     8701 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     8761 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     8821 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     8881 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     8941 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     9001 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     9061 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     9121 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     9181 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     9241 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     9301 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     9361 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     9421 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     9481 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     9541 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     9601 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     9661 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     9721 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     9781 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     9841 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     9901 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     9961 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
    10021 cgaggaagcg gaag
//
