LOCUS       sequence-461674-       10257 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bxb1-GA donor plasmid with divergent 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 10257)
  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 10257)
  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_Divergent
FEATURES             Location/Qualifiers
     source          1..10257
                     /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)."
     misc_feature    1931..2519
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             2037..2048
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     2446..2466
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2523..3218)
                     /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(2798..2817)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3067..3085
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      3215..3224
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3303..3327)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    3372..3390
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3408..3426
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3444..3462
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3480..3498
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3516..3534
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3552..3570
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3588..3606
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     enhancer        3639..4018
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4020..4297
                     /label=chicken beta-actin promoter
     primer_bind     5239..5261
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     5323..5342
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             5374..6117
                     /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             6118..6171
                     /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             6172..6879
                     /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             6904..6924
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6928..6948
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6952..6972
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(7037..7054)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7043..7267
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    7315..7348
                     /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(7356..7372)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7386..7402)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7395..7417)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7615..8070
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7702..7721)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7912..7933
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8087..8106)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8206..8228
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8266..8284)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8352..8456
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8457..8525,8526..9317)
                     /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(8675..8694)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9488..10076
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9977..9996
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10230..10247
                     /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 ctagttaatc accgcatccc cagcactagc gcggggaggc
     1921 ggcccaaagg gcggggaggc ggcccaaagg gagatccgac tcgtctgagg gcgaaggcga
     1981 agacgcggaa gaggccgcag agccggcagc aggccgcggg aaggaaggtc cgctggattg
     2041 agggccgaag ggacgtagca gaaggacgtc ccgcgcagaa tccaggtggc aacacaggcg
     2101 agcagccatg gaaaggacgt cagcttcccc gacaacacca cggaattgtc agtgcccaac
     2161 agccgagccc ctgtccagca gcgggcaagg caggcggcga tgagttccgc cgtggcaata
     2221 gggaggggga aagcgaaagt cccggaaagg agctgacagg tggtggcaat gccccaacca
     2281 gtgggggttg cgtcagcaaa cacagtgcac accacgccac gttgcctgac aacgggccac
     2341 aactcctcat aaagagacag caaccaggat ttatacaagg aggagaaaat gaaagccata
     2401 cgggaagcaa tagcatgata caaaggcatt aaagcagcgt atccacatag cgtaaaagga
     2461 gcaacatagt taagaatacc agtcaatctt tcacaaattt tgtaatccag aggttgattt
     2521 tacttgtaca gctcgtccat gccgccggtg gagtggcggc cctcggagcg ttcgtactgt
     2581 tccaccacgg tgtagtcctc gttgtgggag gtgatgtcca acttgcggtc gacgttgtag
     2641 gcgccgggca tctgcacggg cttcttggcc ttgtaggtgg tcttgaagtc cgccaggtag
     2701 cggccgccgt ccttcaggcg cagggccatc ttaatgtcgc ccttcagcac gccgtcctcg
     2761 gggtacaacc gctcggtgga cgcttcccag cccattgtct tcttctgcat tacggggccg
     2821 tcaggaggga agttggtgcc gcggagcttc accttgtaga tcagggtgcc gtcctccagg
     2881 gaggtgtcct gggtcacggt cacggcgccg ccgtcctcga agttcatcac gcgctcccac
     2941 ttgaagccct cggggaagga ctgcttatag tagtcgggga tgtcggcggg gtgcttggtg
     3001 aaggccctgg agccgtacat gaactgaggg gacaggatgt cccaggagaa gggcaggggg
     3061 ccacccttgg tcaccttcag cttggcggtc tgggtgccct cgtaggggcg gccctcgccc
     3121 tcgccctcga tctcgaactc gtggccgttc atggagccct ccatgtgcac cttgaaccgc
     3181 atgaactcct tgatcactgc ctcgcccttg ctcaccatgg tggcgaattc ttaagtttac
     3241 gagggtagga agtggtacgg aaagttggta taagacaaaa gtgttgtgga attgctccag
     3301 gcgatctgac ggttcactaa acgagctctg cttttatagg cgcccaccgt acacgcctaa
     3361 agcttatacg ttctctatca ctgataggga gtaaactgga tatacgttct ctatcactga
     3421 tagggagtaa actgtagata cgttctctat cactgatagg gagtaaactg gtcatacgtt
     3481 ctctatcact gatagggagt aaactcctta tacgttctct atcactgata gggagtaaag
     3541 tctgcatacg ttctctatca ctgataggga gtaaactctt catacgttct ctatcactga
     3601 tagggagtaa accttaatta actagtcgaa ttaagctcga cattgattat tgactagtta
     3661 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
     3721 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
     3781 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
     3841 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
     3901 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
     3961 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
     4021 cgaggtgagc cccacgttct gcttcactct ccccatctcc cccccctccc cacccccaat
     4081 tttgtattta tttatttttt aattattttg tgcagcgatg ggggcggggg gggggggggg
     4141 gcgcgcgcca ggcggggcgg ggcggggcga ggggcggggc ggggcgaggc ggagaggtgc
     4201 ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct tttatggcga ggcggcggcg
     4261 gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgc gctgccttcg
     4321 ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt
     4381 actcccacag gtgagcgggc gggacggccc ttctcctccg ggctgtaatt agcgcttggt
     4441 ttaatgacgg cttgtttctt ttctgtggct gcgtgaaagc cttgaggggc tccgggaggg
     4501 ccctttgtgc ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg tggggagcgc
     4561 cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg cggggctttg
     4621 tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc ggtgccccgc ggtgcggggg
     4681 gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt gagcaggggg
     4741 tgtgggcgcg tcggtcgggc tgcaaccccc cctgcacccc cctccccgag ttgctgagcg
     4801 cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc gcggggctcg ccgtgccggg
     4861 cggggggtgg cggcmgtacg gggcgtggcg cggggctcgc cgtgccgggc ggggggtggc
     4921 ggcaggtggg ggtgccgggc ggcccccgga gcgccggcgg ctgtcgaggc gcggcgagcc
     4981 gcagccattg ccttttatgg taatcgtgcg agagggcgca gggacttcct ttgtcccaaa
     5041 tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc ctctagcggg cgcggggcga
     5101 agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg tcgccgcgcc
     5161 gccgtcccct tctccctctc cagcctcggg gctgtccgcg gggggacggc tgccttcggg
     5221 ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac cggcggctct agagcctctg
     5281 ctaaccatgt tcatgccttc ttctttttcc tacagctcct gggcaacgtg ctggttattg
     5341 tgctgtctca tcattttggc aaagaatgtc gacatgagcc ggctggacaa gagcaaagtg
     5401 atcaacagcg ccctggaact gctgaacggc gtgggcatcg agggcctgac cacccggaag
     5461 ctggcccaga aactgggcgt ggaacagccc accctgtact ggcacgtgaa gaacaagcgg
     5521 gccctgctgg acgccctgcc catcgagatg ctggaccggc accacaccca cagctgccct
     5581 ctggaaggcg agagctggca ggacttcctg cggaacaacg ccaagagcta cagatgcgcc
     5641 ctgctgagcc accgggacgg cgccaaagtg cacctgggca ccagacccac cgagaagcag
     5701 tacgagacac tggaaaacca gctggccttc ctgtgccagc agggcttcag cctggaaaac
     5761 gccctgtacg ccctgagcgc cgtgggccac ttcaccctgg gctgcgtgct ggaagaacag
     5821 gaacaccagg tcgccaaaga ggaaagagag acacccacca ccgacagcat gccccccctg
     5881 ctgaagcagg ccatcgagct gttcgacaga cagggcgccg agcccgcctt cctgttcggc
     5941 ctggaactga tcatctgcgg cctcgagaag cagctgaagt gcgagagcgg cggacccacc
     6001 gacgccctgg acgacttcga cctggacatg ctgcccgccg atgccctgga tgattttgat
     6061 ctggatatgc tccctgctga cgctctcgat gacttcgatc tcgatatgct gccaggcgag
     6121 ggcaggggaa gtcttctaac atgcggggac gtggaggaaa atcccggccc cgtgtctaag
     6181 ggcgaagagc tgattaagga gaacatgcac atgaagctgt acatggaggg caccgtggac
     6241 aaccatcact tcaagtgcac atccgagggc gaaggcaagc cctacgaggg cacccagacc
     6301 atgagaatca aggtggtcga gggcggccct ctccccttcg ccttcgacat cctggctact
     6361 agcttcctct acggcagcaa gaccttcatc aaccacaccc agggcatccc cgacttcttc
     6421 aagcagtcct tccctgaggg cttcacatgg gagagagtca ccacatacga agacgggggc
     6481 gtgctgaccg ctacccagga caccagcctc caggacggct gcctcatcta caacgtcaag
     6541 atcagagggg tgaacttcac atccaacggc cctgtgatgc agaagaaaac actcggctgg
     6601 gaggccttca ccgagacgct gtaccccgct gacggcggcc tggaaggcag aaacgacatg
     6661 gccctgaagc tcgtgggcgg gagccatctg atcgcaaacg ccaagaccac atatagatcc
     6721 aagaaacccg ctaagaacct caagatgcct ggcgtctact atgtggacta cagactggaa
     6781 agaatcaagg aggccaacaa cgagacctac gtcgagcagc acgaggtggc agtggccaga
     6841 tactgcgacc tccctagcaa actggggcac aagcttaatt ccggactcag atctcgagct
     6901 gatccaaaaa agaagagaaa ggtagatcca aaaaagaaga gaaaggtaga tccaaaaaag
     6961 aagagaaagg taggatccac cggatctaga taactgatca taatcagcca taccacattt
     7021 gaaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     7081 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     7141 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     7201 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     7261 tctatggcta gccgctgcaa gaactcttcc tcacggatcc gaagttccta ttccgaagtt
     7321 cctattcttc aaatagtata ggaacttcag cgctcactgg ccgtcgtttt acaaagggcg
     7381 aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     7441 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     7501 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt
     7561 ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc
     7621 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     7681 ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     7741 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     7801 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     7861 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     7921 tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga
     7981 ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     8041 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg
     8101 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     8161 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     8221 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     8281 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     8341 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     8401 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     8461 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     8521 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     8581 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     8641 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     8701 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     8761 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     8821 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     8881 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     8941 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     9001 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     9061 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     9121 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     9181 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     9241 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     9301 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     9361 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     9421 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     9481 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     9541 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     9601 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     9661 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     9721 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     9781 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     9841 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     9901 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     9961 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
    10021 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
    10081 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
    10141 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
    10201 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaag
//
