LOCUS       sequence-225419-       16228 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The affinity mutant for the F40-based human desmoplakin I tension 
            sensor incorporates a point mutation that exhibits enhanced keratin 
            association..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16228)
  AUTHORS   Price AJ, Cost AL, Ungewiss H, Waschke J, Dunn AR, Grashoff C
  TITLE     Mechanical loading of desmosomes depends on the magnitude and 
            orientation of external stress.
  JOURNAL   Nat Commun. 2018 Dec 11;9(1):5284. doi: 10.1038/s41467-018-07523-0.
  PUBMED    30538252
REFERENCE   2  (bases 1 to 16228)
  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: "Nat 
            Commun."; date: "2018-12-11"; volume: "9(1):5284. doi"; pages: " 
            10.1038/s41467-018-07523-0"
COMMENT     Sequence Label: Desmoplakin I affinity mutant (F40-based tensi...
FEATURES             Location/Qualifiers
     source          1..16228
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        295..607
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    303..321
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    339..357
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    374..392
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    410..428
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    446..464
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    481..499
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    517..535
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     580..604
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             6928..7632
                     /codon_start=1
                     /product="monomeric teal (cyan) fluorescent protein (Ai et 
                     al., 2006)"
                     /label=mTFP1
                     /note="mammalian codon-optimized"
                     /translation="VSKGEETTMGVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDGT
                     NTINLEVKEGAPLPFSYDILTTAFAYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTFE
                     DKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKKTTGWDASTERMYVRDGVLKG
                     DVKHKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYESAV
                     ARNSTDGMDELYK"
     CDS             join(7753..7755,7756..7758,7759..8469)
                     /codon_start=1
                     /product="enhanced YFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7798..7819)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(8059..8078)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8406..8427
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    11364..11485
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11375..11394)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     11429..11448
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             join(12208..12825,12826..12831,12832..12867,12868..12870,
                     12871..12906,12907..12909,12910..12945,12946..12951)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     CDS             12958..13014
                     /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             13015..13614
                     /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(13015..13034)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     13511..13531
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(13669..13686)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    13675..13899
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(14225..14242)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(14396..14984)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(14476..14495)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(15155..15946,15947..16015))
                     /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     15778..15797
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(16016..16120)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggatccctat acagttgaag tcggaagttt acatacactt aagttggagt cattaaaact
       61 cgtttttcaa ctactccaca aatttcttgt taacaaacaa tagttttggc aagtcagtta
      121 ggacatctac tttgtgcatg acacaagtca tttttccaac aattgtttac agacagatta
      181 tttcacttat aattcactgt atcacaattc cagtgggtca gaagtttaca tacactaagt
      241 tcgactcctc tgcagaatgc ggcgatgttt cggtaagggg tccgctatct agacgagttt
      301 actccctatc agtgatagag aacgtatgtc gagtttactc cctatcagtg atagagaacg
      361 atgtcgagtt tactccctat cagtgataga gaacgtatgt cgagtttact ccctatcagt
      421 gatagagaac gtatgtcgag tttactccct atcagtgata gagaacgtat gtcgagttta
      481 tccctatcag tgatagagaa cgtatgtcga gtttactccc tatcagtgat agagaacgta
      541 tgtcgattag gcgtgtacgg tgggcgccta taaaagcaga gctcgtttag tgaaccgtca
      601 gatcgcctgg agccaattcc aactcttttg ccttatacct actttccgta ccacttccta
      661 ccctcgaaag gcctctgcgg ccgcgtcaac agcccctgct ccttgggccc ctccatgcca
      721 tgccgtaatc tctcccaccc gaccaacacc aacacccagc tccgacgcag ctcctctgcg
      781 cccttgccgc cctccgagcc acagctttcc tcccgctcct gcccccggcc cgtcgccgtc
      841 tccgcgctcg cagcggcctc gggagggccc aggtagcgag cagcgacctc gcgagccttc
      901 cgcactcccg cccggttccc cggccgtccg cctatccttg gccccctccg ctttctccgc
      961 gccggcccgc ctcgcttatg cctcggcgct gagccgctct cccgattgcc cgccgacatg
     1021 agctgcaacg gaggctccca cccgcggatc aacactctgg gccgcatgat ccgcgccgag
     1081 tctggcccgg acctgcgcta cgaggtgacc agcggcggcg ggggcaccag caggatgtac
     1141 tattctcggc gcggcgtgat caccgaccag aactcggacg gctactgtca aaccggcacg
     1201 atgtccaggc accagaacca gaacaccatc caggagctgc tgcagaactg ctccgactgc
     1261 ttgatgcgag cagagctcat cgtgcagcct gaattgaagt atggagatgg aatacaactg
     1321 actcggagtc gagaattgga tgagtgtttt gcccaggcca atgaccaaat ggaaatcctc
     1381 gacagcttga tcagagagat gcggcagatg ggccagccct gtgatgctta ccagaaaagg
     1441 cttcttcagc tccaagagca aatgcgagcc ctttataaag ccatcagtgt ccctcgagtc
     1501 cgcagggcca gctccaaggg tggtggaggc tacacttgtc agagtggctc tggctgggat
     1561 gagttcacca aacatgtcac cagtgaatgt ttggggtgga tgaggcagca aagggcggag
     1621 atggacatgg tggcctgggg tgtggacctg gcctcagtgg agcagcacat taacagccac
     1681 cggggcatcc acaactccat cggcgactat cgctggcagc tggacaaaat caaagccgac
     1741 ctgcgcgaga aatctgcgat ctaccagttg gaggaggagt atgaaaacct gctgaaagcg
     1801 tcctttgaga ggatggatca cctgcgacag ctgcagaaca tcattcaggc cacgtccagg
     1861 gagatcatgt ggatcaatga ctgcgaggag gaggagctgc tgtacgactg gagcgacaag
     1921 aacaccaaca tcgctcagaa acaggaggcc ttctccatac gcatgagtca actggaagtt
     1981 aaagaaaaag agctcaataa gctgaaacaa gaaagtgacc aacttgtcct caatcagcat
     2041 ccagcttcag acaaaattga ggcctatatg gacactctgc agacgcagtg gagttggatt
     2101 cttcagatca ccaagtgcat tgatgttcat ctgaaagaaa atgctgccta ctttcagttt
     2161 tttgaagagg cgcagtctac tgaagcatac ctgaaggggc tccaggactc catcaggaag
     2221 aagtacccct gcgacaagaa catgcccctg cagcacctgc tggaacagat caaggagctg
     2281 gagaaagaac gagagaaaat ccttgaatac aagcgtcagg tgcagaactt ggtaaacaag
     2341 tctaagaaga ttgtacagct gaagcctcgt aacccagact acagaagcaa taaacccatt
     2401 attctcagag ctctctgtga ctacaaacaa gatcagaaaa tcgtgcataa gggggatgag
     2461 tgtatcctga aggacaacaa cgagcgcagc aagtggtacg tgacgggccc gggaggcgtt
     2521 gacatgcttg ttccctctgt ggggctgatc atccctcctc cgaacccact ggccgtggac
     2581 ctctcttgca agattgagca gtactacgaa gccatcttgg ctctgtggaa ccagctctac
     2641 atcaacatga agagcctggt gtcctggcac tactgcatga ttgacataga gaagatcagg
     2701 gccatgacaa tcgccaagct gaaaacaatg cggcaggaag attacatgaa gacgatagcc
     2761 gaccttgagt tacattacca agagttcatc agaaatagcc aaggctcaga gatgtttgga
     2821 gatgatgaca agcggaaaat acagtctcag ttcaccgatg cccagaagca ttaccagacc
     2881 ctggtcattc agctccctgg ctatccccag caccagacag tgaccacaac tgaaatcact
     2941 catcatggaa cctgccaaga tgtcaaccat aataaagtaa ttgaaaccaa cagagaaaat
     3001 gacaagcaag aaacatggat gctgatggag ctgcagaaga ttcgcaggca gatagagcac
     3061 tgcgagggca ggatgactct caaaaacctc cctctagcag accaggggtc ttctcaccac
     3121 atcacagtga aaattaacga gcttaagagt gtgcagaatg attcacaagc aattgctgag
     3181 gttctcaacc agcttaaaga tatgcttgcc aacttcagag gttctgaaaa gtactgctat
     3241 ttacagaatg aagtatttgg actatttcag aaactggaaa atatcaatgg tgttacagat
     3301 ggctacttaa atagcttatg cacagtaagg gcactgctcc aggctattct ccaaacagaa
     3361 gacatgttaa aggtttatga agccaggctc actgaggagg aaactgtctg cctggacctg
     3421 gataaagtgg aagcttaccg ctgtggactg aagaaaataa aaaatgactt gaacttgaag
     3481 aagtcgttgt tggccactat gaagacagaa ctacagaaag cccagcagat ccactctcag
     3541 acttcacagc agtatccact ttatgatctg gacttgggca agttcggtga aaaagtcaca
     3601 cagctgacag accgctggca aaggatagat aaacagatcg actttagatt atgggacctg
     3661 gagaaacaaa tcaagcaatt gaggaattat cgtgataact atcaggcttt ctgcaagtgg
     3721 ctctatgatg ctaaacgccg ccaggattcc ttagaatcca tgaaatttgg agattccaac
     3781 acagtcatgc ggtttttgaa tgagcagaag aacttgcaca gtgaaatatc tggcaaacga
     3841 gacaaatcag aggaagtaca aaaaattgct gaactttgcg ccaattcaat taaggattat
     3901 gagctccagc tggcctcata cacctcagga ctggaaactc tgctgaacat acctatcaag
     3961 aggaccatga ttcagtcccc ttctggggtg attctgcaag aggctgcaga tgttcatgct
     4021 cggtacattg aactacttac aagatctgga gactattaca ggttcttaag tgagatgctg
     4081 aagagtttgg aagatctgaa gctgaaaaat accaagatcg aagttttgga agaggagctc
     4141 agactggccc gagatgccaa ctcggaaaac tgtaataaga acaaattcct ggatcagaac
     4201 ctgcagaaat accaggcaga gtgttcccag ttcaaagcga agcttgcgag cctggaggag
     4261 ctgaagagac aggctgagct ggatgggaag tcggctaagc aaaatctaga caagtgctac
     4321 ggccaaataa aagaactcaa tgagaagatc acccgactga cttatgagat tgaagatgaa
     4381 aagagaagaa gaaaatctgt ggaagacaga tttgaccaac agaagaatga ctatgaccaa
     4441 ctgcagaaag caaggcaatg tgaaaaggag aaccttggtt ggcagaaatt agagtctgag
     4501 aaagccatca aggagaagga gtacgagatt gaaaggttga gggttctact gcaggaagaa
     4561 ggcacccgga agagagaata tgaaaatgag ctggcaaagg taagaaacca ctataatgag
     4621 gagatgagta atttaaggaa caagtatgaa acagagatta acattacgaa gaccaccatc
     4681 aaggagatat ccatgcaaaa agaggatgat tccaaaaatc ttagaaacca gcttgataga
     4741 ctttcaaggg aaaatcgaga tctgaaggat gaaattgtca ggctcaatga cagcatcttg
     4801 caggccactg agcagcgaag gcgagctgaa gaaaacgccc ttcagcaaaa ggcctgtggc
     4861 tctgagataa tgcagaagaa gcagcatctg gagatagaac tgaagcaggt catgcagcag
     4921 cgctctgagg acaatgcccg gcacaagcag tccctggagg aggctgccaa gaccattcag
     4981 gacaaaaata aggagatcga gagactcaaa gctgagtttc aggaggaggc caagcgccgc
     5041 tgggaatatg aaaatgaact gagtaaggta agaaacaatt atgatgagga gatcattagc
     5101 ttaaaaaatc agtttgagac cgagatcaac atcaccaaga ccaccatcca ccagctcacc
     5161 atgcagaagg aagaggatac cagtggctac cgggctcaga tagacaatct cacccgagaa
     5221 aacaggagct tatctgaaga aataaagagg ctgaagaaca ctctaaccca gaccacagag
     5281 aatctcagga gggtggaaga agacatccaa cagcaaaagg ccactggctc tgaggtgtct
     5341 cagaggaaac agcagctgga ggttgagctg agacaagtca ctcagatgcg aacagaggag
     5401 agcgtaagat ataagcaatc tcttgatgat gctgccaaaa ccatccagga taaaaacaag
     5461 gagatagaaa ggttaaaaca actgatcgac aaagaaacaa atgaccggaa atgcctggaa
     5521 gatgaaaacg cgagattaca aagggtccag tatgacctgc agaaagcaaa cagtagtgcg
     5581 acggagacaa taaacaaact gaaggttcag gagcaagaac tgacacgcct gaggatcgac
     5641 tatgaaaggg tttcccagga gaggactgtg aaggaccagg atatcacgcg gttccagaac
     5701 tctctgaaag agctgcagct gcagaagcag aaggtggaag aggagctgaa tcggctgaag
     5761 aggaccgcgt cagaagactc ctgcaagagg aagaagctgg aggaagagct ggaaggcatg
     5821 aggaggtcgc tgaaggagca agccatcaaa atcaccaacc tgacccagca gctggagcag
     5881 gcatccattg ttaagaagag gagtgaggat gacctccggc agcagaggga cgtgctggat
     5941 ggccacctga gggaaaagca gaggacccag gaagagctga ggaggctctc ttctgaggtc
     6001 gaggccctga ggcggcagtt actccaggaa caggaaagtg tcaaacaagc tcacttgagg
     6061 aatgagcatt tccagaaggc gatagaagat aaaagcagaa gcttaaatga aagcaaaata
     6121 gaaattgaga ggctgcagtc tctcacagag aacctgacca aggagcactt gatgttagaa
     6181 gaagaactgc ggaacctgag gctggagtac gatgacctga ggagaggacg aagcgaagcg
     6241 gacagtgata aaaatgcaac catcttggaa ctaaggagcc agctgcagat cagcaacaac
     6301 cggaccctgg aactgcaggg gctgattaat gatttacaga gagagaggga aaatttgaga
     6361 caggaaattg agaaattcca aaagcaggct ttagaggcat ctaataggat tcaggaatca
     6421 aagaatcagt gtactcaggt ggtacaggaa agagagagcc ttctggtgaa aatcaaagtc
     6481 ctggagcaag acaaggcaag gctgcagagg ctggaggatg agctgaatcg tgcaaaatca
     6541 actctagagg cagaaaccag ggtgaaacag cgcctggagt gtgagaaaca gcaaattcag
     6601 aatgacctga atcagtggaa gactcaatat tcccgcaagg aggaggctat taggaagata
     6661 gaatcggaaa gagaaaagag tgagagagag aagaacagtc ttaggagtga gatcgaaaga
     6721 ctccaagcag agatcaagag aattgaagag aggtgcaggc gtaagctgga ggattctacc
     6781 agggagacac agtcacagtt agaaacagaa cgctcccgat atcagaggga gattgataaa
     6841 ctcagacagc gcttaattaa gggaagcggc ggaaccggat cgactagcgg gggtagcggg
     6901 ggaagcacgg ggggaggaac tggcgccgtg agcaagggcg aggagaccac aatgggcgta
     6961 atcaagcccg acatgaagat caagctgaag atggagggca acgtgaatgg ccacgccttc
     7021 gtgatcgagg gcgagggcga gggcaagccc tacgacggca ccaacaccat caacctggag
     7081 gtgaaggagg gagcccccct gcccttctcc tacgacattc tgaccaccgc gttcgcctac
     7141 ggcaacaggg ccttcaccaa gtaccccgac gacatcccca actacttcaa gcagtccttc
     7201 cccgagggct actcttggga gcgcaccatg accttcgagg acaagggcat cgtgaaggtg
     7261 aagtccgaca tctccatgga ggaggactcc ttcatctacg agatacacct caagggcgag
     7321 aacttccccc ccaacggccc cgtgatgcag aagaagacca ccggctggga cgcctccacc
     7381 gagaggatgt acgtgcgcga cggcgtgctg aagggcgacg tcaagcacaa gctgctgctg
     7441 gagggcggcg gccaccaccg cgttgacttc aagaccatct acagggccaa gaaggcggtg
     7501 aagctgcccg actatcactt tgtggaccac cgcatcgaga tcctgaacca cgacaaggac
     7561 tacaacaagg tgaccgttta cgagagcgcc gtggcccgca actccaccga cggcatggac
     7621 gagctgtaca aggggccagg tggtgcaggg ccaggtggtg cagggccagg tggtgcaggg
     7681 ccaggtggtg cagggcccgg tggtgcaggt ccaggtggtg caggtccagg tggtgcaggt
     7741 ccaggtggtg ctatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     7801 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     7861 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     7921 ccctggccca ccctcgtgac caccttcggc tacggcctgc agtgcttcgc ccgctacccc
     7981 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     8041 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     8101 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     8161 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     8221 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     8281 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     8341 cccgacaacc actacctgag ctaccagtcc aaactgagca aagaccccaa cgagaagcgc
     8401 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     8461 ctgtacaagg gcaccggagg tggcactagc ggaggctcag gcggatcgac ctccggaact
     8521 ggaggttccg gtagcggccg cccatatggg tcccatcgag agacccagac tgagtgtgag
     8581 tggaccgttg acacctccaa gctggtgttt gatgggctga ggaagaaggt gacagcaatg
     8641 cagctctatg agtgtcagct gatcgacaaa acaaccttgg acaaactatt gaaggggaag
     8701 aagtcagtgg aagaagttgc ttctgaaatc cagccattcc ttcggggtgc aggatctatc
     8761 gctggagcat ctgcttctcc taaggaaaaa tactctttgg tagaggccaa gagaaagaaa
     8821 ttaatcagcc cagaatccac agtcatgctt ctggaggccc aggcagctac aggtggtata
     8881 attgatcccc atcggaatga gaagctgact gtcgacagtg ccatagctcg ggacctcatt
     8941 gacttcgatg accgtcagca gatatatgca gcagaaaaag ctatcactgg ttttgatgat
     9001 ccattttcag gcaagacagt atctgtttca gaagccatca agaaaaattt gattgataga
     9061 gaaaccggaa tgcgcctgct ggaagcccag attgcttcag ggggtgtagt agaccctgtg
     9121 aacagtgtct ttttgccaaa agatgtcgcc ttggcccggg ggctgattga tagagatttg
     9181 tatcgatccc tgaatgatcc ccgagatagt cagaaaaact ttgtggatcc agtcaccaaa
     9241 aagaaggtca gttacgtgca gctgaaggaa cggtgcagaa tcgaaccaca tactggtctg
     9301 ctcttgcttt cagtacagaa gagaagcatg tccttccaag gaatcagaca acctgtgacc
     9361 gtcactgagc tagtagattc tggtatattg agaccgtcca ctgtcaatga actggaatct
     9421 ggtcagattt cttatgacga ggttggtgag agaattaagg acttcctcca gggttcaagc
     9481 tgcatagcag gcatatacaa tgagaccaca aaacagaagc ttggcattta tgaggccatg
     9541 aaaattggct tagtccgacc tggtactgct ctggagttgc tggaagccca agcagctact
     9601 ggctttatag tggatcctgt tagcaacttg aggttaccag tggaggaagc ctacaagaga
     9661 ggtctggtgg gcattgagtt caaagagaag ctcctgtctg cagaacgagc tgtcactggg
     9721 tataatgatc ctgaaacagg aaacatcatc tctttgttcc aagccatgaa taaggaactc
     9781 atcgaaaagg gccacggtat tcgcttatta gaagcacaga tcgcaaccgg ggggatcatt
     9841 gacccaaagg agagccatcg tttaccagtt gacatagcat ataagagggg ctatttcaat
     9901 gaggaactca gtgagattct ctcagatcca agtgatgata ccaaaggatt ttttgacccc
     9961 aacactgaag aaaatcttac ctatctgcaa ctaaaagaaa gatgcattaa ggatgaggaa
    10021 acagggctct gtcttctgcc tctgaaagaa aagaagaaac aggtgcagac atcacaaaag
    10081 aataccctca ggaagcgtag agtggtcata gttgacccag aaaccaataa agaaatgtct
    10141 gttcaggagg cctacaagaa gggcctaatt gattatgaaa ccttcaaaga actgtgtgag
    10201 caggaatgtg aatgggaaga aataaccatc acgggatcag atggctccac cagggtggtc
    10261 ctggtagata gaaagacagg cagtcagtat gatattcaag atgctattga caagggcctt
    10321 gttgacagga agttctttga tcagtaccga tccggcagcc tcagcctcac tcaatttgct
    10381 gacatgatct ccttgaaaaa tggtgtcggc accagcagca gcatgggcag tggtgtcagc
    10441 gatgatgttt ttagcagctc ccgacatgaa tcagtaagta agatttccac catatccagc
    10501 gtcaggaatt taaccataag gagcagctct ttttcagaca ccctggaaga atcgagcccc
    10561 attgcagcca tctttgacac agaaaacctg gagaaaatct ccattacaga aggtatagag
    10621 cggggcatcg ttgacagcat cacgggtcag aggcttctgg aggctcaggc ctgcacaggt
    10681 ggcatcatcc acccaaccac gggccagaag ctgtcacttc aggacgcagt ctcccagggt
    10741 gtgattgacc aagacatggc caccaggctg aagcctgctc agaaagcctt cataggcttc
    10801 gagggtgtga agggaaagaa gaagatgtca gcagcagagg cagtgaaaga aaaatggctc
    10861 ccgtatgagg ctggccagcg cttcctggag ttccagtacc tcacgggagg tcttgttgac
    10921 ccggaagtgc atgggaggat aagcaccgaa gaagccatcc ggaaggggtt catagatggc
    10981 cgcgccgcac agaggctgca agacaccagc agctatgcca aaatcctgac ctgccccaaa
    11041 accaaattaa aaatatccta taaggatgcc ataaatcgct ccatggtaga agatatcact
    11101 gggctgcgcc ttctggaagc cgcctccgtg tcgtccaagg gcttacccag cccttacaac
    11161 atgtcttcgg ctccggggtc ccgctccggc tcccgctcgg gatctcgctc cggatctcgc
    11221 tccgggtccc gcagtgggtc ccggagagga ggctttgacg ccacagggaa ttcttcctac
    11281 tcttattcct actcatttag cagtagttct attgggcact aggctagcgg cctgtcaggc
    11341 caagcttcca tcgatagaca tgataagata cattgatgag tttggacaaa ccacaacaag
    11401 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
    11461 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
    11521 tcagggggag atgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtaccta
    11581 ggcgagaccc tgtctcacaa aataaagtaa gcccggactg agtgcggaaa ggcgggcctg
    11641 gcgggtctgg tctccccatg cgggccacca gaggccctgc agccttcagt cgcttgaagg
    11701 ggtaatggcg cttccactca caaacatggc ggacagagcg tgtgaacgag atgaacagcc
    11761 cctcaaaaat atggccgccg aggctggacg gccgtgcccc agcagcaccg cctccgcgcc
    11821 ccacgtgatc tctcgccggg cacagcgctg accgcggagg tccaaccgga agaatgtccg
    11881 gattggacat tcggaagagg gcccgccttc cctggggaat ctctgcgcac gcgcagaacg
    11941 cttcgaccaa tgaaaacaca ggaagccgtc cgcgcaaccg cgttgcgtca cttctgccgc
    12001 ccctgtttca aggtatatag ccgtagacgg aacttcgcct ttctctcggc cttagcgcca
    12061 tttttttggg tgagtgtttt ttggttcctg cgttgggatt ccgtgtacaa tccatagaca
    12121 tctgacctcg gcacttagca tcatcacagc aaactaactg tagcctttct ctctttccct
    12181 gtagaaacct ctgcacctga ggccaccatg tctagactgg acaagagcaa agtcataaac
    12241 ggcgctctgg aattactcaa tggagtcggt atcgaaggcc tgacgacaag gaaactcgct
    12301 caaaagctgg gagttgagca gcctaccctg tactggcacg tgaagaacaa gcgggccctg
    12361 ctcgatgccc tgccaatcga gatgctggac aggcatcata cccacttctg ccccctggaa
    12421 ggcgagtcat ggcaagactt tctgcggaac aacgccaagt cattccgctg tgctctcctc
    12481 tcacatcgcg acggggctaa agtgcatctc ggcacccgcc caacagagaa acagtacgaa
    12541 accctggaaa atcagctcgc gttcctgtgt cagcaaggct tctccctgga gaacgcactg
    12601 tacgctctgt ccgccgtggg ccactttaca ctgggctgcg tattggagga acaggagcat
    12661 caagtagcaa aagaggaaag agagacacct accaccgatt ctatgccccc acttctgaga
    12721 caagcaattg agctgttcga ccggcaggga gccgaacctg ccttcctttt cggcctggaa
    12781 ctaatcatat gtggcctgga gaaacagcta aagtgcgaaa gcggcgggcc ggccgacgcc
    12841 cttgacgatt ttgacttaga catgctccca gccgatgccc ttgacgactt tgaccttgat
    12901 atgctgcctg ctgacgctct tgacgatttt gaccttgaca tgctccccgg gtccggcgct
    12961 actaacttca gcctgctgaa gcaggctggt gacgtcgagg agaatcctgg tcccatgacc
    13021 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc
    13081 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac
    13141 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
    13201 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc
    13261 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
    13321 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg
    13381 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc
    13441 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc
    13501 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
    13561 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgattcgaa
    13621 ggcctgtcgt gaagcttggg gatcaattct ctagagctcg ctgatcagcc tcgactgtgc
    13681 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
    13741 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
    13801 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
    13861 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
    13921 gctgggccta ctagctactc gggacccctt accgaaacat cgccgcattc tgcagaggag
    13981 tcgagtgtat gtaaacttct gacccactgg gaatgtgatg aaagaaataa aagctgaaat
    14041 gaatcattct ctctactatt attctgatat ttcacattct taaaataaag tggtgatcct
    14101 aactgaccta agacagggaa tttttactag gattaaatgt caggaattgt gaaaaagtga
    14161 gtttaaatgt atttggctaa ggtgtatgta aacttccgac ttcaactgta tagggatccg
    14221 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
    14281 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
    14341 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
    14401 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
    14461 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
    14521 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
    14581 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
    14641 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
    14701 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
    14761 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
    14821 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
    14881 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
    14941 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
    15001 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
    15061 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
    15121 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
    15181 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
    15241 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
    15301 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
    15361 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
    15421 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
    15481 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
    15541 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
    15601 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
    15661 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
    15721 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
    15781 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
    15841 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
    15901 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
    15961 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
    16021 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
    16081 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
    16141 cacctgatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggaaa
    16201 ttgtaagcgt taatattttg ttaaaatt
//
