LOCUS       sequence-485741-       11109 bp    DNA     circular SYN 10-SEP-2025
DEFINITION  Bxb1-GA donor plasmid with downstream tandem syntax for all-in-one 
            grazoprevir-inducible expression of ETV2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11109)
  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 11109)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 10, 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-GZV-SynZiFTR_ETV2_Down-Tandem
FEATURES             Location/Qualifiers
     source          1..11109
                     /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)."
     regulatory      2353..2362
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    3386..3974
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3439..3459)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3857..3868)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     enhancer        4031..4410
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4412..4685
                     /label=chicken beta-actin promoter
     intron          4686..5702
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     5626..5648
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     5710..5729
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             6320..6385
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             6392..6412
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7262..7288
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             7514..7885
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALLSQISS"
     polyA_signal    7895..8119
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(8208..8224)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8238..8254)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8247..8269)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8467..8922
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8554..8573)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8764..8785
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8939..8958)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9058..9080
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(9118..9136)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9204..9308
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9309..9377,9378..10169)
                     /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(9527..9546)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10340..10928
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10829..10848
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11082..11099
                     /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 aagcggacga ttgagtaaca ctgtatctag
     1921 tctacctaca gggctcggcg tagccgatgt cgcgcggata gcgtcaacct cttcggcgta
     1981 gccgatgtcg cgcaatcagt tctcccgagg tcggcgtagc cgatgtcgcg cagagccgtg
     2041 cttccaatac ggcgtagccg atgtcgcgcc gggtttcgta acaatcgcat gaggattcgc
     2101 aacgccttcg gcgtagccga tgtcgcgctc ccgtctcagt aaaggtcggc gtagccgatg
     2161 tcgcgcaatc ggactgcctt cgtacggcgt agccgatgtc gcgcgtatca gtcgcctcgg
     2221 aacggcgtag ccgatgtcgc gcattcgtaa gaggctcact ctcccttaca cggagtggat
     2281 aggatcctct agagggtata taatgggggc cactagtcta ctaccagagc tcatcgctag
     2341 cgctaccgga tcgccaccat ggacctgtgg aactgggatg aggcatcccc acaggaagtg
     2401 cctccaggga acaagctggc agggcttgaa ggagccaaat taggcttctg tttccctgat
     2461 ctggcactcc aaggggacac gccgacagcg acagcagaga catgctggaa aggtacaagc
     2521 tcatccctgg caagcttccc acagctggac tggggctccg cgttactgca cccagaagtt
     2581 ccatgggggg cggagcccga ctctcaggct cttccgtggt ccggggactg gacagacatg
     2641 gcgtgcacag cctgggactc ttggagcggc gcctcgcaga ccctgggccc cgcccctctc
     2701 ggcccgggcc ccatccccgc cgccggctcc gaaggcgccg cgggccagaa ctgcgtcccc
     2761 gtggcgggag aggccacctc gtggtcgcgc gcccaggccg ccgggagcaa caccagctgg
     2821 gactgttctg tggggcccga cggcgatacc tactggggca gtggcctggg cggggagccg
     2881 cgcacggact gtaccatttc gtggggcggg cccgcgggcc cggactgtac cacctcctgg
     2941 aacccggggc tgcatgcggg tggcaccacc tctttgaagc ggtaccagag ctcagctctc
     3001 accgtttgct ccgaaccgag cccgcagtcg gaccgtgcca gtttggctcg atgccccaaa
     3061 actaaccacc gaggtcccat tcagctgtgg cagttcctcc tggagctgct ccacgacggg
     3121 gcgcgtagca gctgcatccg ttggactggc aacagccgcg agttccagct gtgcgacccc
     3181 aaagaggtgg ctcggctgtg gggcgagcgc aagagaaagc cgggcatgaa ttacgagaag
     3241 ctgagccggg gccttcgcta ctactatcgc cgcgacatcg tgcgcaagag cggggggcga
     3301 aagtacacgt accgcttcgg gggccgcgtg cccagcctag cctatccgga ctgtgcggga
     3361 ggcggacggg gagcagagac acaataatca acctctggat tacaaaattt gtgaaagatt
     3421 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     3481 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     3541 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     3601 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     3661 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     3721 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     3781 gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     3841 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     3901 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     3961 ggccgcctcc ccgccctttg ggccgcctcc ccgcgctagt gctggggatg cggtgggctc
     4021 tatgggtctc gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
     4081 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
     4141 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     4201 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     4261 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     4321 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     4381 atctacgtat tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact
     4441 ctccccatct cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt
     4501 tgtgcagcga tgggggcggg gggggggggc gcgcgccagg cggggcgggg cggggcgagg
     4561 ggcggggcgg ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa
     4621 agtttccttt tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc
     4681 gggcgggagt cgctgcgcgc tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc
     4741 gcccgccccg gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt
     4801 ctcctccggg ctgtaattag cgcttggttt aatgacggct tgtttctttt ctgtggctgc
     4861 gtgaaagcct tgaggggctc cgggagggcc ctttgtgcgg ggggagcggc tcggggggtg
     4921 cgtgcgtgtg tgtgtgcgtg gggagcgccg cgtgcggctc cgcgctgccc ggcggctgtg
     4981 agcgctgcgg gcgcggcgcg gggctttgtg cgctccgcag tgtgcgcgag gggagcgcgg
     5041 ccgggggcgg tgccccgcgg tgcggggggg gctgcgaggg gaacaaaggc tgcgtgcggg
     5101 gtgtgtgcgt gggggggtga gcagggggtg tgggcgcgtc ggtcgggctg caaccccccc
     5161 tgcacccccc tccccgagtt gctgagcacg gcccggcttc gggtgcgggg ctccgtacgg
     5221 ggcgtggcgc ggggctcgcc gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg
     5281 gggcggggcc gcctcgggcc ggggagggct cgggggaggg gcgcggcggc ccccggagcg
     5341 ccggcggctg tcgaggcgcg gcgagccgca gccattgcct tttatggtaa tcgtgcgaga
     5401 gggcgcaggg acttcctttg tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc
     5461 gcaccccctc tagcgggcgc ggggcgaagc ggtgcggagc cggcaggaag gaaatgggcg
     5521 gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag cctcggggct
     5581 gtccgcgggg ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc
     5641 gtgtgaccgg cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac
     5701 agctcctggg caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattccgat
     5761 cgcgccacct gccgccacca tgtctagacc cggagagcgc ccattccagt gtcggatttg
     5821 catgcggaac ttttcgagaa gacacggcct ggacagacat acccgtactc atacaggtga
     5881 aaaacccttt cagtgtcgga tctgtatgcg aaatttctcc gaccacagca gcctgaagag
     5941 acatctacgt acccacaccg gcagccagaa gccatttcag tgtcggatct gtatgcggaa
     6001 cttctccgtg agacacaacc tgaccagaca tctacgtacg cacaccggag agaagccatt
     6061 ccaatgccga atatgcatgc gcaacttcag tgaccacagc aacctgagca gacacctaaa
     6121 aacccacacc ggttcccaga agccatttca gtgtcggatc tgtatgcgga acttctccca
     6181 gcgcagcagc ctggtgagac atctacgtac gcacaccgga gagaagccat tccaatgccg
     6241 aatatgcatg cgcaacttca gtgagagcgg ccacctgaag agacacctgc gtacgcacct
     6301 gaggggatcc acctgcaggg actacaaaga ccatgacggt gattataaag atcatgacat
     6361 cgattacaag gatgacgatg acaagatggc ccccaagaaa aagaggaagg tgggcattca
     6421 cggggtgccg ggtggactcg agggaggcgg tggaagcggc ggtaccgagg acgtggtgtg
     6481 ctgccactca atctacggca agaagaaggg tgatatcgac acctaccgat acataggctc
     6541 ttccgggaca ggctgcgtgg tcatagtggg caggatcgtc ttgtccggat ccggcactag
     6601 tgcgcccatc acggcgtacg cccagcagac gagaggcctc ctagggtgta taatcaccag
     6661 cctgactggc cgggacaaaa accaagtgga gggtgaggtc cagatcgtgt caactgctac
     6721 ccaaaccttc ctggcaacgt gcatcaatgg ggtatgctgg gcagtctacc acggggccgg
     6781 aacgaggacc atcgcatcac ccaagggtcc tgtcatccag atgtatacca atgtggacca
     6841 agaccttgtg ggctggcccg ctcctcaagg ttcccgctca ttgacaccct gtacctgcgg
     6901 ctcctcggac ctttacctgg tcacgaggca cgccgatgtc attcccgtgc gccggcgagg
     6961 tgatagcagg ggtagcctgc tttcgccccg gcccatttcc tacttgaaag gctcctcggg
     7021 gggtccgctg ttgtgccccg cgggacacgc cgtgggccta ttcagggccg cggtgtgcac
     7081 ccgtggagtg gctaaagcgg tggactttat ccctgtggag aacctagaga caaccatgag
     7141 atccccggtg ttcacggaca actcctctcc accagcagtc accctgacgc acccaatcac
     7201 caaaatcgat agggaggttc tctaccagga gttcgatgag atggaagagt gctctcagca
     7261 ctatccctac gatgtgcccg attacgctgg aggcggtgga agcggcggta ccgatgagtt
     7321 tcccaccatg gtgtttcctt ctgggcagat cagccaggcc tcggccttgg ccccggcccc
     7381 tccccaagtc ctgccccagg ctccagcccc tgcccctgct ccagccatgg tatcagctct
     7441 ggcccaggcc ccagcccctg tcccagtcct agccccaggc cctcctcagg ctgtggcccc
     7501 acctgccccc aagcccaccc aggctgggga aggaacgctg tcagaggccc tgctgcagct
     7561 gcagtttgat gatgaagacc tgggggcctt gcttggcaac agcacagacc cagctgtgtt
     7621 cacagacctg gcatccgtcg acaactccga gtttcagcag ctgctgaacc agggcatacc
     7681 tgtggccccc cacacaactg agcccatgct gatggagtac cctgaggcta taactcgcct
     7741 agtgacaggg gcccagaggc cccccgaccc agctcctgct ccactggggg ccccggggct
     7801 ccccaatggc ctcctttcag gagatgaaga cttctcctcc attgcggaca tggacttctc
     7861 agccctgctg agtcagatca gctcccaatt gtaactgtgc cttctagttg ccagccatct
     7921 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     7981 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     8041 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     8101 gatgcggtgg gctctatggc tagccgctgc aagaactctt cctcacggat ccgaagttcc
     8161 tattccgaag ttcctattct tcaaatagta taggaacttc agcgctcact ggccgtcgtt
     8221 ttacaaaggg cgaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     8281 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     8341 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg
     8401 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac
     8461 catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     8521 tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc
     8581 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     8641 gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta
     8701 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     8761 atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg
     8821 atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa
     8881 aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta
     8941 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     9001 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     9061 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
     9121 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     9181 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     9241 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     9301 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     9361 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     9421 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     9481 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     9541 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     9601 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     9661 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     9721 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     9781 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     9841 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     9901 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     9961 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
    10021 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
    10081 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
    10141 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
    10201 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
    10261 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
    10321 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
    10381 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
    10441 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
    10501 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
    10561 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
    10621 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
    10681 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
    10741 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
    10801 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
    10861 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
    10921 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
    10981 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
    11041 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
    11101 aagcggaag
//
