LOCUS       sequence-428949-       13174 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PiggyBac vector for stable co-expression of mCherry-LgBiT and 
            EGFP-Vinculin-PILATeS (700 pM) in mammalian cells. Requires 
            transposase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13174)
  AUTHORS   Zhong BL, Elliot JM, Wang P, Li H, Hall RN, Wang B, Prakash M, Dunn 
            AR
  TITLE     Split Luciferase Molecular Tension Sensors for Bioluminescent 
            Readout of Mechanical Forces in Biological Systems.
  JOURNAL   ACS Sens. 2024 Jul 26;9(7):3489-3495. doi: 
            10.1021/acssensors.3c02664. Epub 2024 Jul 7.
  PUBMED    38973210
REFERENCE   2  (bases 1 to 13174)
  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: 
            "10.1021/acssensors.3c02664"; journalName: "ACS Sens"; date: 
            "2024-07-26- 26"; volume: "9"; issue: "7"; pages: "3489-3495"
COMMENT     Sequence Label: mCh-LgBiT-2A-EGFP-Vin-PILATeS
FEATURES             Location/Qualifiers
     source          1..13174
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1010..1030
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1012..1030
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     repeat_region   1078..1112
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     polyA_signal    1400..1521
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(1568..2167)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MAEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1651..1671)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     promoter        complement(2186..2233)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(2242..3453)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(2386..2405)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     enhancer        3475..3854
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3856..4131
                     /label=chicken beta-actin promoter
     intron          4132..5147
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     5071..5093
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     5155..5174
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      5224..5233
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5230..5937
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(5375..5393)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5643..5662
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     5837..5860
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             6433..6489
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             6499..6552
                     /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(6553..6555,6556..7266)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(6553..6555,6556..7266)
                     /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(6595..6616)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6856..6875)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7203..7224
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    10967..11088
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   11274..11336
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(11371..11390)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(11372..11390)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(11400..11417)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(11400..11416)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11409..11431)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      11558..12013
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11645..11664)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11855..11876
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        12039..12143
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(12144..12212,12213..13004)
                     /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(12362..12381)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     1021 tcactaaagg gaacaaaagc tggagctcca ccgcggggcg cgccgcagct agattaaccc
     1081 tagaaagata gtctgcgtaa aattgacgca tgcattcttg aaatattgct ctctctttct
     1141 aaatagcgcg aatccgtcgc tgtgcattta ggacatctca gtcgccgctt ggagctcccg
     1201 tgaggcgtgc ttgtcaatgc ggtaagtgtc actgattttg aactataaca accgcgtgag
     1261 tcaaaatgac gcatgattat cttttacgtg acttttaaga tttaactcat acgataatta
     1321 tattgttatt tcgtgttcta cttacgtgat aacttattat atatatattt tcttgttata
     1381 gatgggaatt cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     1441 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     1501 ataagctgca ataaacaagt ttcgaggtcg agtgtcagtc ctgctcctcg gccacgaagt
     1561 gcttagctca ggcaccgggc ttgcgggtca tgcaccaggt gcgcggtcct tcgggcacct
     1621 cgacgtcggc ggtgacggtg aagccgagcc gctcgtagaa ggggaggttg cggggcgcgg
     1681 aggtctccag gaaggcgggc accccggcgc gctcggccgc ctccactccg gggagcacga
     1741 cggcgctgcc cagacccttg ccctggtggt cgggcgagac gccgacggtg gccaggaacc
     1801 acgcgggctc cttgggccgg tgcggcgcca ggaggccttc catctgttgc tgcgcggcca
     1861 gccgggaacc gctcaactcg gccatgcgcg ggccgatctc ggcgaacacc gcccccgctt
     1921 cgacgctctc cggcgtggtc cagaccgcca ccgcggcgcc gtcgtccgcg acccacacct
     1981 tgccgatgtc gagcccgacg cgcgtgagga agagttcttg cagctcggtg acccgctcga
     2041 tgtggcggtc cgggtcgacg gtgtggcgcg tggcggggta gtcggcgaac gcggcggcga
     2101 gggtgcgtac ggcccggggg acgtcgtcgc gggtggcgag gcgcaccgtg ggcttgtact
     2161 cggccatggt ttagttcctc accttgtcgt attatactat gccgatatac tatgccgatg
     2221 attaattgtc aacacggtcc ggtctaacaa aaaagccaaa aacggccaga atttagcgga
     2281 caatttacta gtctaacact gaaaattaca tattgaccca aatgattaca tttcaaaagg
     2341 tgcctaaaaa acttcacaaa acacactcgc caaccccgag cgcatagttc aaaaccggag
     2401 cttcagctac ttaagaagat aggtacataa aaccgaccaa agaaactgac gcctcactta
     2461 tccctcccct caccagaggt ccggcgcctg tcgattcagg agagcctacc ctaggcccga
     2521 accctgcgtc ctgcgacgga gaaaagccta ccgcacacct accggcaggt ggccccaccc
     2581 tgcattataa gccaacagaa cgggtgacgt cacgacacga cgagggcgcg cgctcccaaa
     2641 ggtacgggtg cactgcccaa cggcaccgcc ataactgccg cccccgcaac agacgacaaa
     2701 ccgagttctc cagtcagtga caaacttcac gtcagggtcc ccagatggtg ccccagccca
     2761 tctcacccga ataagagctt tcccgcatta gcgaaggcct caagaccttg ggttcttgcc
     2821 gcccaccatg ccccccacct tgtttcaacg acctcacagc ccgcctcaca agcgtcttcc
     2881 attcaagact cgggaacagc cgccattttg ctgcgctccc cccaaccccc agttcagggc
     2941 aaccttgctc gcggacccag actacagccc ttggcggtct ctccacacgc ttccgtccca
     3001 ccgagcggcc cggcggccac gaaagccccg gccagcccag cagcccgcta ctcaccaagt
     3061 gacgatcaca gcgatccaca aacaagaacc gcgacccaaa tcccggctgc gacggaacta
     3121 gctgtgccac acccggcgcg tccttatata atcatcggcg ttcaccgccc cacggagatc
     3181 cctccgcaga atcgccgaga agggactact tttcctcgcc tgttccgctc tctggaaaga
     3241 aaaccagtgc cctagagtca cccaagtccc gtcctaaaat gtccttctgc tgatactggg
     3301 gttctaaggc cgagtcttat gagcagcggg ccgctgtcct gagcgtccgg gcggaaggat
     3361 caggacgctc gctgcgccct tcgtctgacg tggcagcgct cgccgtgagg aggggggcgc
     3421 ccgcgggggg cgccaaaacc cggcgcggag gccagatctc ccgatctcga ggtcgacatt
     3481 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     3541 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     3601 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     3661 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     3721 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     3781 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     3841 tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc atctcccccc
     3901 cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca gcgatggggg
     3961 cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg gggcggggcg
     4021 aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt tccttttatg
     4081 gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc gggagtcgct
     4141 gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc gccccggctc
     4201 tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt
     4261 aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga aagccttgag
     4321 gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg cgtgtgtgtg
     4381 tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg ctgcgggcgc
     4441 ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg gggcggtgcc
     4501 ccgcggtgcg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg tgcgtggggg
     4561 ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac ccccctcccc
     4621 gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt ggcgcggggc
     4681 tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg gggccgcctc
     4741 gggccgggga gggctcgggg gaggggcgcg gcggcccccg gagcgccggc ggctgtcgag
     4801 gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg cagggacttc
     4861 ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg
     4921 ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag ggccttcgtg
     4981 cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg cggggggacg
     5041 gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct
     5101 ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg
     5161 tgctggttat tgtgctgtct catcattttg gcaaagaatt cctcgacggt atcgataagc
     5221 ttggccacca tggtgagcaa gggcgaggag gataacatgg ccatcatcaa ggagttcatg
     5281 cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag
     5341 ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac caagggcggc
     5401 cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac
     5461 gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag
     5521 tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc
     5581 ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac
     5641 ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct cctccgagcg gatgtacccc
     5701 gaggacggcg ccctgaaggg cgagatcaag cagaggctga agctgaagga cggcggccac
     5761 tacgacgccg aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc
     5821 tacaacgtca acatcaagct ggacatcacc tcccacaacg aggactacac catcgtggaa
     5881 cagtacgagc gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaaggga
     5941 agcggagtct tcactctgga agatttcgta ggtgactggg agcaaacagc agcatacaat
     6001 cttgatcaag tgttggagca agggggggta agctctctcc tccaaaactt ggctgtctca
     6061 gttactccaa ttcaacgaat tgtacggtca ggggaaaatg cactcaagat tgacattcat
     6121 gtcatcattc cttatgaggg tctgagtgcc gatcagatgg cccagatcga ggaggttttt
     6181 aaggtagtgt atcccgtgga tgatcatcac tttaaggtaa ttttgcccta cggaactctg
     6241 gtaattgacg gagtgacccc caatatgctc aattattttg gtaggcccta tgaaggtatt
     6301 gctgtcttcg acggaaaaaa aataacagtt actgggacac tctggaatgg taataagatt
     6361 atagacgaac gcttgatcac tcctgacggc agtatgttgt tccgcgttac cataaatggg
     6421 tctggaagcg gagccacgaa cttctctctg ttaaagcaag caggagacgt ggaagaaaac
     6481 cccggtcctg gaagcggaga gggcagggga agtcttctaa catgcgggga cgtggaggaa
     6541 aatcccggcc ccgtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     6601 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     6661 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     6721 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     6781 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     6841 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     6901 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     6961 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     7021 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     7081 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     7141 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     7201 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     7261 tacaagtccg gactcggagg aagcggaggt ggaggaagcg gaggtggagg aagcggaggt
     7321 atgcccgtct tccacacgcg caccatcgag agcatcttgg agcccgtggc tcagcagatc
     7381 tcccacctgg tcatcatgca cgaggagggg gaggtagacg gcaaggccat cccggacctc
     7441 accgcccccg tgtcggccgt gcaggccgct gtcagcaacc tggtgcgggt tggaaaagaa
     7501 actgtgcaga caacagaaga ccagatcttg aaaagggata tgccaccagc attcatcaaa
     7561 gtagagaatg cctgcaccaa gctcgttcga gcagcccaga tgctgcaagc agatccttat
     7621 tcagtaccag ctcgtgacta cctaattgat ggatcaagag gcatcctttc tggaacatca
     7681 gacttacttc tgacatttga tgaagcggag gtccgtaaaa tcatccgtgt ctgcaaagga
     7741 atattggaat atctgactgt ggcagaagta gtagagacta tggaggattt ggtgacatat
     7801 acaaagaatc tagggccagg aatgacaaag atggcgaaaa tgatcgatga gagacaacag
     7861 gaattaactc atcaggaaca tagggttatg ctggtgaact ccatgaatac tgtgaaggag
     7921 ctattgcctg tacttatttc agctatgaag atctttgtaa ccacaaaaaa cactaaaagc
     7981 cagggaatag aagaggcctt gaaaaatcgc aatttcacag tagagaaaat gagtgctgag
     8041 ataaatgaaa taatccgtgt attacaactc acttcctggg atgaagatgc ctgggccagc
     8101 aaggacactg aagccatgaa aagagcttta gccctaatag attcaaagat gaaccaggca
     8161 aaaggctggc tgagagatcc aaacgcacct ccaggggatg ctggtgagca agcaataagg
     8221 cagatccttg atgaagctgg aaaagcagga gaattgtgtg caggcaaaga acgcagggag
     8281 attctgggaa catgcaaaac tttgggccaa atgactgatc aacttgctga tctccgagct
     8341 agaggacagg gtgctacacc gatggcaatg cagaaggcac agcaggtgtc acaaggcctg
     8401 gatttgctca ctgcaaaagt ggagaatgca gcccggaaat tggaggccat gacaaactct
     8461 aagcaggcta ttgcaaagaa gattgatgct gctcagaact ggcttgcgga tcccaacggc
     8521 ggaagtgaag gagaagaaca cattcgagga attatgtctg aagcaaggaa agttgcagaa
     8581 ttgtgtgagg agcctaaaga aagagatgat atccttcgct ccttggggga aatctctgct
     8641 ctgacagcta agctgtcaga tctgcgacga catgggaaag gcgactctcc tgaggcccgt
     8701 gcattggcca agcaaatagc tacatcactt cagaacttac agtccaaaac aaacagagct
     8761 gtagcaaata ctaggccagt taaagctgct gtccatttgg agggcaagat tgagcaagcc
     8821 cagaggtgga tagataatcc tacagttgat gatcggggag taggccaggc agcaattcgg
     8881 ggtttggttg cagaaggtcg tcgtctagcc aatgtcatga tgggacctta tcgtcaggac
     8941 ctgcttgcca aatgtgaccg tgtagaccag ctggctgctc agcttgctga ccttgcagca
     9001 agaggggaag gagagtctcc tcaggctagg gcaattgctg ctcagcttca ggactccctg
     9061 aaggatctca aagcacggat gcaagaagca atgacccagg aggtgtctga tgttttcagt
     9121 gacactacaa cccctattaa gttgttagca gtagcagcca ctgctccttc tgatactccc
     9181 aatagagaag aggtgtttga agaaagagca gcaaattttg aaaaccatgc tgctagactg
     9241 ggagcaacag cagaaaaagc agctgcagtt ggaactgcta ataaaactac tgtggaaggc
     9301 attcaggcaa cagtcaaatc tgcaagggag cttacaccac aggtagtatc ggctgctcga
     9361 atcctcctga gaaatcctgg aaatcaagct gcttatgagc attttgagac aatgaaaaac
     9421 caatggattg ataatgtaga aaagatgaca gggctggtgg atgaggccat cgataccaag
     9481 tctctgttgg atgcatcaga agaggctatt aagaaggatc ttgataaatg caaagttgca
     9541 atggccaata tgcaacctca gatgctggta gctggagcca ccagcattgc tagacgagcg
     9601 aaccgcatcc tgcttgtggc aaaacgggag gttgaaaatt cagaagaccc taaattcagg
     9661 gaggctgtta aagcagcttc tgatgagctg agcaaaacca tatcaccgat ggtaatggat
     9721 gctaaagctg tagcaggaaa tatctctgat cctggtttgc agaagagttt cttggattct
     9781 ggatacagga ttctgggagc tgtggccaaa gtcagagaag cctttcagcc tcaggagcca
     9841 gactttcccc ctcctcctcc tgacctagag catctccatc tgactgatga gcttgctcct
     9901 ccaaaaccac cacttccaga aggtgaggtt cccccaccca gaccaccacc ccctgaagaa
     9961 aaagatgagg tcgagatgga aactctgcat attaccaaaa ccatgaaaaa cattgaagtg
    10021 ccggaaacca aaaccgcgag ctttgaatgc gaagtgagcc attttaacgt gccgagcatg
    10081 tggctgaaaa acggcgtgga aattgaaatg agcgaaaaat ttaaaattgt ggtgcagggc
    10141 aaactgcatc agctgattat tatgaacacc agcaccgaag atagcgcgga atataccttt
    10201 gtgtgcggcg ttagtggctg gcgtctgttt aagaaaatta gcaacgatca ggtgagcgcg
    10261 accctgaccg tgactgcggc cgcggagttc ccagagcaga aagcaggaga agctattaat
    10321 cagcccatga tgatggctgc taggcagttg catgacgagg ctcggaaatg gtctagcaag
    10381 ggtaacgaca tcattgctgc tgctaaacga atggcgctgc tcatggctga gatgtcacgc
    10441 ctggtgcgag gaggcagcgg aaacaagcgt gccctcatcc agtgtgcaaa agatattgct
    10501 aaggcatcgg atgaagtcac tcgattggcc aaagaggtgg ccaagcaatg tactgataaa
    10561 cgcattagaa caaacctctt acaggtctgt gagcgaatcc caaccatcag cacgcagctc
    10621 aaaattcttt ccacagtgaa agctaccatg ctgggcagga ctaacatcag cgatgaagaa
    10681 tcagaacagg caactgagat gttggttcat aacgcccaga acctcatgca gtctgtgaag
    10741 gaaactgtga gagaagctga agcagcatcc attaagataa gaacagatgc cggattcact
    10801 ctgcgctggg tcagaaagac cccatggtat cagtagttta attaagcggc cgcgactcta
    10861 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
    10921 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
    10981 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
    11041 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaag gaattcgata
    11101 aaagttttgt tactttatag aagaaatttt gagtttttgt ttttttttaa taaataaata
    11161 aacataaata aattgtttgt tgaatttatt attagtatgt aagtgtaaat ataataaaac
    11221 ttaatatcta ttcaaattaa taaataaacc tcgatataca gaccgataaa acacatgcgt
    11281 caattttacg catgattatc tttaacgtac gtcacaatat gattatcttt ctagggttaa
    11341 tctagctgcg gcgcgccggt acccaattcg ccctatagtg agtcgtatta cgcgcgctca
    11401 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
    11461 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
    11521 ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta
    11581 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
    11641 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
    11701 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
    11761 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
    11821 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
    11881 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
    11941 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
    12001 acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
    12061 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
    12121 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
    12181 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
    12241 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
    12301 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
    12361 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
    12421 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
    12481 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
    12541 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
    12601 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
    12661 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
    12721 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
    12781 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
    12841 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
    12901 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
    12961 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
    13021 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
    13081 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
    13141 cgtcagaccc cgtagaaaag atcaaaggat cttc
//
