LOCUS       sequence-426920-        9383 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For integration of MYOD1 AroPERFECT C T2A mEGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9383)
  AUTHORS   Naderi J, Magalhaes AP, Kibar G, Stik G, Zhang Y, Mackowiak SD, 
            Wieler HM, Rossi F, Buschow R, Christou-Kent M, Alcoverro-Bertran M,
            Graf T, Vingron M, Hnisz D
  TITLE     An activity-specificity trade-off encoded in human transcription 
            factors.
  JOURNAL   Nat Cell Biol. 2024 Aug;26(8):1309-1321. doi: 
            10.1038/s41556-024-01411-0. Epub 2024 Jul 5.
  PUBMED    38969762
REFERENCE   2  (bases 1 to 9383)
  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"; doi: 
            "10.1038/s41556-024-01411-0"; journalName: "Nat Cell Biol"; date: 
            "2024-08"; volume: "26"; issue: "8"; pages: "1309-1321"
COMMENT     Sequence Label: pPB-TetON-mEGFP-MYOD1_AroPERFECT-C
FEATURES             Location/Qualifiers
     source          1..9383
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   147..209
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        658..1036
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    666..684
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    702..720
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    738..756
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    774..792
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    810..828
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    846..864
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    882..900
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     945..969
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1049..1058
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1058..1123
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2087..2140
                     /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             join(2147..2149,2150..2860)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(2147..2149,2150..2860)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2189..2210)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2450..2469)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2797..2818
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2947..2966)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2991..3039
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        3270..3481
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             3494..3762
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             3796..4539
                     /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             4540..4593
                     /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             4594..5193
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4594..4613)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5090..5110
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5226..5347
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5263..5282)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5317..5336
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   6110..6144
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(6635..6651)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6635..6651)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6648..6670)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6659..6675
                     /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(6683..6689,6690..6707,6708..6713))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6728..6749
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6866..6883)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7037..7625)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7117..7136)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7796..8587,8588..8656))
                     /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     8419..8438
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8657..8761)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(8787..9242)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8924..8945)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     9136..9155
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     join(9369..9383,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(9383,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgag cgcgcctcgt tcattcacgt ttttgaaccc gtggaggacg
       61 ggcagactcg cggtgcaaat gtgttttaca gcgtgatgga gcagatgaag atgctcgaca
      121 cgctgcagaa cacgcagcta gattaaccct agaaagataa tcatattgtg acgtacgtta
      181 aagataatca tgtgtaaaat tgacgcatgt gttttatcgg tctgtatatc gaggtttatt
      241 tattaatttg aatagatatt aagttttatt atatttacac ttacatacta ataataaatt
      301 caacaaacaa tttatttatg tttatttatt tattaaaaaa aacaaaaact caaaatttct
      361 tctataaagt aacaaaactt ttatgaggga cagccccccc ccaaagcccc cagggatgta
      421 attacgtccc tcccccgcta gggggcagca gcgagccgcc cggggctccg ctccggtccg
      481 gcgctccccc cgcatccccg agccggcagc gtgcggggac agcccgggca cggggaaggt
      541 ggcacgggat cgctttcctc tgaacgcttc tcgctgctct ttgagcctgc agacacctgg
      601 ggggatacgg ggaaaaggcc tccacggcca ctagtctttc gtcttcaaga attcctcgag
      661 tttactccct atcagtgata gagaacgtat gaagagttta ctccctatca gtgatagaga
      721 acgtatgcag actttactcc ctatcagtga tagagaacgt ataaggagtt tactccctat
      781 cagtgataga gaacgtatga ccagtttact ccctatcagt gatagagaac gtatctacag
      841 tttactccct atcagtgata gagaacgtat atccagttta ctccctatca gtgatagaga
      901 acgtataagc tttaggcgtg tacggtgggc gcctataaaa gcagagctcg tttagtgaac
      961 cgtcagatcg cctggagcaa ttccacaaca cttttgtctt ataccaactt tccgtaccac
     1021 ttcctaccct cgtaaaggtc tagagctagc caccatggac tacaaagacc atgacggtga
     1081 ttataaagat catgacatcg attacaagga tgacgatgac aagatggagc ttctctcacc
     1141 gcccctgagg gatgtggatt tgaccgctcc tgatggatcc ttgtgtagct tcgctacgac
     1201 agatgacttt tacgatgatc cttgctttga tagtcctgat cttcggttct ttgaggattt
     1261 ggatcctaga ctcatgcatg ttggagccct tcttaagcct gaggaacatt cccattttcc
     1321 ggctgccgtc catcccgcgc ccggagctag ggaagatgaa cacgttagag ccccttctgg
     1381 acatcatcaa gccgggagat gtttgttgtg ggcgtgtaaa gcctgtaaaa ggaaaactac
     1441 aaatgctgat aggcgcaaag cggctacaat gagggaacga cggcgactta gtaaggttaa
     1501 cgaagctttc gaaaccctga aacgatgtac ttccagtaac cccaatcaac gactgcctaa
     1561 agttgaaatt ctcagaaatg ctattcggta cattgaagga cttcaagcct tgttgcgaga
     1621 tcaagatgct gcaccccctg gagctgccgc tgctgctcct gggcctctcc cacccgggcg
     1681 aggcggtgaa tggcatagtg gggatagtga tgcctcatct cccaggtcta attgtagttt
     1741 tgatgggatg atggattctg ggcccccttc cggggcgcga cgcagataca attgtgaggg
     1801 tgctaatgaa gccccttccg agcctcgacc tggctataaa tccgcagctg tcagctctct
     1861 ggattgtctc agctctattg tttatgaaag aataagcaca gaatcacccg ccgcacctgc
     1921 tctgctcttg tatgcagatg tccccagcga atctccccca agaagacagg aagccgcata
     1981 tgcgccatcc gaaggcgaat cctctggtga tcctacgcaa tccccgtacg acgccgcccc
     2041 gcagtgccct gcgggtgcga accccaaccc gatataccag gtgctcgagg gcagaggaag
     2101 tcttctaaca tgcggtgacg tggaggagaa tcccggccct agtaccgtga gcaagggcga
     2161 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     2221 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     2281 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     2341 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     2401 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     2461 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     2521 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     2581 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
     2641 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     2701 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     2761 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     2821 cgccgccggg atcactctcg gcatggacga gctgtacaag accggttgac cggttagtaa
     2881 tgagtttaaa cgggggagga agaccggttg accggttagt aatgagttta aacgggggag
     2941 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     3001 aagacagaat aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     3061 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     3121 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     3181 tcggggcggc aggccctgcc atagcagatc tgcgaaatcg gatccagctg aatctaagtg
     3241 gatctgcgat cgctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga
     3301 gaagttgggg ggaggggtcg gcaattgaac gggtgcctag agaaggtggc gcggggtaaa
     3361 ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta
     3421 tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca
     3481 gctgaagctt cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg
     3541 ccatccacgc cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc
     3601 gtccgccgtc taggtaagtt taaagctcag gtcgagaccg ggcctttgtc cggcgctccc
     3661 ttggagccta cctagactca gccggctctc cacgctttgc ctgaccctgc ttgctcaact
     3721 ctacgtcttt gtttcgtttt ctgttctgcg ccgttacaga tctaagctgt gaccggcgcc
     3781 tactctagag ccgccatgtc tagactggac aagagcaaag tcataaactc tgctctggaa
     3841 ttactcaatg gagtcggtat cgaaggcctg acgacaagga aactcgctca aaagctggga
     3901 gttgagcagc ctaccctgta ctggcacgtg aagaacaagc gggccctgct cgatgccctg
     3961 ccaatcgaga tgctggacag gcatcatacc cactcctgcc ccctggaagg cgagtcatgg
     4021 caagactttc tgcggaacaa cgccaagtca taccgctgtg ctctcctctc acatcgcgac
     4081 ggggctaaag tgcatctcgg cacccgccca acagagaaac agtacgaaac cctggaaaat
     4141 cagctcgcgt tcctgtgtca gcaaggcttc tccctggaga acgcactgta cgctctgtcc
     4201 gccgtgggcc actttacact gggctgcgta ttggaggaac aggagcatca agtagcaaaa
     4261 gaggaaagag agacacctac caccgattct atgcccccac ttctgaaaca agcaattgag
     4321 ctgttcgacc ggcagggagc cgaacctgcc ttccttttcg gcctggaact aatcatatgt
     4381 ggcctggaga aacagctaaa gtgcgaaagc ggcgggccga ccgacgccct tgacgatttt
     4441 gacttagaca tgctcccagc cgatgccctt gacgactttg accttgatat gctgcctgct
     4501 gacgctcttg acgattttga ccttgacatg ctccccgggg agggcagagg aagtcttcta
     4561 acatgcggtg acgtggagga gaatcccggc cctatgaccg agtacaagcc cacggtgcgc
     4621 ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac
     4681 taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg
     4741 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac
     4801 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc
     4861 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg
     4921 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc
     4981 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag
     5041 gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc
     5101 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc
     5161 acctggtgca tgacccgcaa gcccggtgcc tgaaatcaac ctctggatta caaaatttgt
     5221 gggtaaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     5281 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     5341 tatcttatca tgtctggaat tgactcaaat gatgtcaatt agtctatcag aagctcatct
     5401 ggtctccctt ccgggggaca agacatccct gtttaatatt taaacagcag tgttcccaaa
     5461 ctgggttctt atatcccttg ctctggtcaa ccaggttgca gggtttcctg tcctcacagg
     5521 aacgaagtcc ctaaagaaac agtggcagcc aggtttagcc ccggaattga ctggattcct
     5581 tttttagggc ccattggtat ggctttttcc ccgtatcccc ccaggtgtct gcaggctcaa
     5641 agagcagcga gaagcgttca gaggaaagcg atcccgtgcc accttccccg tgcccgggct
     5701 gtccccgcac gctgccggct cggggatgcg gggggagcgc cggaccggag cggagccccg
     5761 ggcggctcgc tgctgccccc tagcggggga gggacgtaat tacatccctg ggggctttgg
     5821 gggggggctg tccctgatat ctataacaag aaaatatata tataataagt tatcacgtaa
     5881 gtagaacatg aaataacaat ataattatcg tatgagttaa atcttaaaag tcacgtaaaa
     5941 gataatcatg cgtcattttg actcacgcgg tcgttatagt tcaaaatcag tgacacttac
     6001 cgcattgaca agcacgcctc acgggagctc caagcggcga ctgagatgtc ctaaatgcac
     6061 agcgacggat tcgcgctatt tagaaagaga gagcaatatt tcaagaatgc atgcgtcaat
     6121 tttacgcaga ctatctttct agggttaatc tagctgcatc aggatcatat cgtcgggtct
     6181 tttttccggc tcagtcatcg cccaagctgg cgctatctgg gcatcgggga ggaagaagcc
     6241 cgtgcctttt cccgcgaggt tgaagcggca tggaaagagt ttgccgagga tgactgctgc
     6301 tgcattgacg ttgagcgaaa acgcacgttt accatgatga ttcgggaagg tgtggccatg
     6361 cacgccttta acggtgaact gttcgttcag gccacctggg ataccagttc gtcgcggctt
     6421 ttccggacac agttccggat ggtcagcccg aagcgcatca gcaacccgaa caataccggc
     6481 gacagccgga actgccgtgc cggtgtgcag attaatgaca gcggtgcggc gctgggatat
     6541 tacgtcagcg aggacgggta tcctggctgg atgccgcaga aatggacatg gataccccgt
     6601 gagttacccg gcgggcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     6661 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     6721 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     6781 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     6841 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     6901 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     6961 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     7021 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     7081 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     7141 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     7201 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     7261 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     7321 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     7381 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     7441 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     7501 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     7561 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     7621 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     7681 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     7741 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     7801 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     7861 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     7921 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     7981 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     8041 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     8101 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     8161 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     8221 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     8281 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     8341 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     8401 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     8461 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     8521 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     8581 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     8641 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     8701 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     8761 gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc
     8821 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     8881 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     8941 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     9001 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     9061 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     9121 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     9181 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     9241 gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     9301 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     9361 agggttttcc cagtcacgac gtt
//
