LOCUS       sequence-461776-       10198 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bxb1-GA donor plasmid with downstream 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 10198)
  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 10198)
  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_Down-Tandem
FEATURES             Location/Qualifiers
     source          1..10198
                     /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)."
     protein_bind    1900..1918
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1936..1954
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1972..1990
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2008..2026
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2044..2062
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2080..2098
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2116..2134
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2179..2203
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2282..2291
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2288..2983
                     /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(2421..2439)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2689..2708
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2987..3575
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3040..3060)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3458..3469)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     enhancer        3632..4011
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4013..4287
                     /label=chicken beta-actin promoter
     primer_bind     5180..5202
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     5264..5283
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             5315..6058
                     /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             6059..6112
                     /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             6113..6820
                     /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             6845..6865
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6869..6889
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6893..6913
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(6978..6995)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6984..7208
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    7256..7289
                     /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(7297..7313)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7327..7343)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7336..7358)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7556..8011
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7643..7662)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7853..7874
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8028..8047)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8147..8169
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8207..8225)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8293..8397
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8398..8466,8467..9258)
                     /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(8616..8635)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9429..10017
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9918..9937
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10171..10188
                     /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 aaggtttact ccctatcagt gatagagaac
     1921 gtatgaagag tttactccct atcagtgata gagaacgtat gcagacttta ctccctatca
     1981 gtgatagaga acgtataagg agtttactcc ctatcagtga tagagaacgt atgaccagtt
     2041 tactccctat cagtgataga gaacgtatct acagtttact ccctatcagt gatagagaac
     2101 gtatatccag tttactccct atcagtgata gagaacgtat aagctttagg cgtgtacggt
     2161 gggcgcctat aaaagcagag ctcgtttagt gaaccgtcag atcgcctgga gcaattccac
     2221 aacacttttg tcttatacca actttccgta ccacttccta ccctcgtaaa cttaagaatt
     2281 cgccaccatg gtgagcaagg gcgaggcagt gatcaaggag ttcatgcggt tcaaggtgca
     2341 catggagggc tccatgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc
     2401 ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttctc
     2461 ctgggacatc ctgtcccctc agttcatgta cggctccagg gccttcacca agcaccccgc
     2521 cgacatcccc gactactata agcagtcctt ccccgagggc ttcaagtggg agcgcgtgat
     2581 gaacttcgag gacggcggcg ccgtgaccgt gacccaggac acctccctgg aggacggcac
     2641 cctgatctac aaggtgaagc tccgcggcac caacttccct cctgacggcc ccgtaatgca
     2701 gaagaagaca atgggctggg aagcgtccac cgagcggttg taccccgagg acggcgtgct
     2761 gaagggcgac attaagatgg ccctgcgcct gaaggacggc ggccgctacc tggcggactt
     2821 caagaccacc tacaaggcca agaagcccgt gcagatgccc ggcgcctaca acgtcgaccg
     2881 caagttggac atcacctccc acaacgagga ctacaccgtg gtggaacagt acgaacgctc
     2941 cgagggccgc cactccaccg gcggcatgga cgagctgtac aagtaaaatc aacctctgga
     3001 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     3061 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     3121 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     3181 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     3241 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     3301 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     3361 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     3421 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     3481 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     3541 gacgagtcgg atctcccttt gggccgcctc cccgcccttt gggccgcctc cccgcgctag
     3601 tgctggggat gcggtgggct ctatgggtct cgacattgat tattgactag ttattaatag
     3661 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
     3721 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     3781 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     3841 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
     3901 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
     3961 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtcgaggtg
     4021 agccccacgt tctgcttcac tctccccatc tcccccccct ccccaccccc aattttgtat
     4081 ttatttattt tttaattatt ttgtgcagcg atgggggcgg gggggggggg gcgcgcgcca
     4141 ggcggggcgg ggcggggcga ggggcggggc ggggcgaggc ggagaggtgc ggcggcagcc
     4201 aatcagagcg gcgcgctccg aaagtttcct tttatggcga ggcggcggcg gcggcggccc
     4261 tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgc gctgccttcg ccccgtgccc
     4321 cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt actcccacag
     4381 gtgagcgggc gggacggccc ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg
     4441 cttgtttctt ttctgtggct gcgtgaaagc cttgaggggc tccgggaggg ccctttgtgc
     4501 ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg tggggagcgc cgcgtgcggc
     4561 tccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg cggggctttg tgcgctccgc
     4621 agtgtgcgcg aggggagcgc ggccgggggc ggtgccccgc ggtgcggggg gggctgcgag
     4681 gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt gagcaggggg tgtgggcgcg
     4741 tcggtcgggc tgcaaccccc cctgcacccc cctccccgag ttgctgagcg cggcccggct
     4801 tcgggtgcgg ggctccgtac ggggcgtggc gcggggctcg ccgtgccggg cggggggtgg
     4861 cggcaggtgg gggtgccggg cggcccccgg agcgccggcg gctgtcgagg cgcggcgagc
     4921 cgcagccatt gccttttatg gtaatcgtgc gagagggcgc agggacttcc tttgtcccaa
     4981 atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc cctctagcgg gcgcggggcg
     5041 aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg gccttcgtgc gtcgccgcgc
     5101 cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc ggggggacgg ctgccttcgg
     5161 gggggacggg gcagggcggg gttcggcttc tggcgtgtga ccggcggctc tagagcctct
     5221 gctaaccatg ttcatgcctt cttctttttc ctacagctcc tgggcaacgt gctggttatt
     5281 gtgctgtctc atcattttgg caaagaatgt cgacatgagc cggctggaca agagcaaagt
     5341 gatcaacagc gccctggaac tgctgaacgg cgtgggcatc gagggcctga ccacccggaa
     5401 gctggcccag aaactgggcg tggaacagcc caccctgtac tggcacgtga agaacaagcg
     5461 ggccctgctg gacgccctgc ccatcgagat gctggaccgg caccacaccc acagctgccc
     5521 tctggaaggc gagagctggc aggacttcct gcggaacaac gccaagagct acagatgcgc
     5581 cctgctgagc caccgggacg gcgccaaagt gcacctgggc accagaccca ccgagaagca
     5641 gtacgagaca ctggaaaacc agctggcctt cctgtgccag cagggcttca gcctggaaaa
     5701 cgccctgtac gccctgagcg ccgtgggcca cttcaccctg ggctgcgtgc tggaagaaca
     5761 ggaacaccag gtcgccaaag aggaaagaga gacacccacc accgacagca tgccccccct
     5821 gctgaagcag gccatcgagc tgttcgacag acagggcgcc gagcccgcct tcctgttcgg
     5881 cctggaactg atcatctgcg gcctcgagaa gcagctgaag tgcgagagcg gcggacccac
     5941 cgacgccctg gacgacttcg acctggacat gctgcccgcc gatgccctgg atgattttga
     6001 tctggatatg ctccctgctg acgctctcga tgacttcgat ctcgatatgc tgccaggcga
     6061 gggcagggga agtcttctaa catgcgggga cgtggaggaa aatcccggcc ccgtgtctaa
     6121 gggcgaagag ctgattaagg agaacatgca catgaagctg tacatggagg gcaccgtgga
     6181 caaccatcac ttcaagtgca catccgaggg cgaaggcaag ccctacgagg gcacccagac
     6241 catgagaatc aaggtggtcg agggcggccc tctccccttc gccttcgaca tcctggctac
     6301 tagcttcctc tacggcagca agaccttcat caaccacacc cagggcatcc ccgacttctt
     6361 caagcagtcc ttccctgagg gcttcacatg ggagagagtc accacatacg aagacggggg
     6421 cgtgctgacc gctacccagg acaccagcct ccaggacggc tgcctcatct acaacgtcaa
     6481 gatcagaggg gtgaacttca catccaacgg ccctgtgatg cagaagaaaa cactcggctg
     6541 ggaggccttc accgagacgc tgtaccccgc tgacggcggc ctggaaggca gaaacgacat
     6601 ggccctgaag ctcgtgggcg ggagccatct gatcgcaaac gccaagacca catatagatc
     6661 caagaaaccc gctaagaacc tcaagatgcc tggcgtctac tatgtggact acagactgga
     6721 aagaatcaag gaggccaaca acgagaccta cgtcgagcag cacgaggtgg cagtggccag
     6781 atactgcgac ctccctagca aactggggca caagcttaat tccggactca gatctcgagc
     6841 tgatccaaaa aagaagagaa aggtagatcc aaaaaagaag agaaaggtag atccaaaaaa
     6901 gaagagaaag gtaggatcca ccggatctag ataactgatc ataatcagcc ataccacatt
     6961 tgaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     7021 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     7081 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     7141 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     7201 ctctatggct agccgctgca agaactcttc ctcacggatc cgaagttcct attccgaagt
     7261 tcctattctt caaatagtat aggaacttca gcgctcactg gccgtcgttt tacaaagggc
     7321 gaattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     7381 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     7441 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt
     7501 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg
     7561 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     7621 cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     7681 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     7741 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg
     7801 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     7861 ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga tttataaggg
     7921 attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     7981 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct
     8041 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     8101 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     8161 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     8221 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     8281 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     8341 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     8401 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     8461 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     8521 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     8581 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     8641 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     8701 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     8761 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     8821 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     8881 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     8941 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     9001 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     9061 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     9121 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     9181 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     9241 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     9301 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     9361 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     9421 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     9481 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     9541 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     9601 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     9661 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     9721 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     9781 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     9841 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     9901 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     9961 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
    10021 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
    10081 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
    10141 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaag
//
