LOCUS       sequence-69233-e       10138 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Destination plasmid (Gateway cloning) for cumate-inducible 
            expression of HA-tagged fusion protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10138)
  AUTHORS   Kaczmarczyk A, Vorholt JA, Francez-Charlot A
  TITLE     Cumate-inducible gene expression system for sphingomonads and other 
            Alphaproteobacteria.
  JOURNAL   Appl Environ Microbiol. 2013 Nov;79(21):6795-802. doi: 
            10.1128/AEM.02296-13. Epub 2013 Aug 30.
  PUBMED    23995928
REFERENCE   2  (bases 1 to 10138)
  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: 
            "10.1128/AEM.02296-13"; journalName: "Appl Environ Microbiol"; date:
            "2013-11"; volume: "79"; issue: "21"; pages: "6795-802"
COMMENT     Sequence Label: pQHD
FEATURES             Location/Qualifiers
     source          1..10138
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      11..642
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      1105..1693
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1755..2366)
                     /codon_start=1
                     /gene="cymR from Pseudomonas putida"
                     /product="cumate repressor (Mullick et al., 2006)"
                     /label=CymR
                     /translation="MSPKRRTQAERAMETQGKLIAAALGVLREKGYAGFRIADVPGAAG
                     VSRGAQSHHFPTKLELLLATFEWLYEQITERSRARLAKLKPEDDVIQQMLDDAAEFFLD
                     DDFSISLDLIVAADRDPALREGIQRTVERNRFVVEDMWLGVLVSRGLSRDDAEDILWLI
                     FNSVRGLAVRSLWQKDKERFERVRNSTLEIARERYAKFKR"
     protein_bind    2443..2470
                     /label=CuO
                     /bound_moiety="CymR"
                     /note="CymR-binding P2 operator sequence from the p-cmt 
                     operon of Pseudomonas putida (Mullick et al., 2006)"
     protein_bind    2513..2540
                     /label=CuO
                     /bound_moiety="CymR"
                     /note="CymR-binding P2 operator sequence from the p-cmt 
                     operon of Pseudomonas putida (Mullick et al., 2006)"
     CDS             2565..2591
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    2610..2734
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(2759..2764,2765..2782,2783..2789)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2843..3502
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2909..2928)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      3555..4143
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4297..4314
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             4776..5081
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     4960..4979
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(5122..5246)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             5288..5353
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(5425..5442)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5425..5441)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5434..5456)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5650..5669)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5769..5791
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5829..5847)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             6633..7832
                     /codon_start=1
                     /gene="tetA"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
                     GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
                     VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
                     FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
                     QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
                     LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
                     AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
                     "
     CDS             complement(8063..9211)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     CDS             complement(9483..9854)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL*AYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            9887..9996
                     /note="incP origin of transfer"
ORIGIN
        1 gaccctttcc gacgctcacc gggctggttg ccctcgccgc tgggctggcg gccgtctatg
       61 gccctgcaaa cgcgccagaa acgccgtcga agccgtgtgc gagacaccgc ggccgccggc
      121 gttgtggata cctcgcggaa aacttggccc tcactgacag atgaggggcg gacgttgaca
      181 cttgaggggc cgactcaccc ggcgcggcgt tgacagatga ggggcaggct cgatttcggc
      241 cggcgacgtg gagctggcca gcctcgcaaa tcggcgaaaa cgcctgattt tacgcgagtt
      301 tcccacagat gatgtggaca agcctgggga taagtgccct gcggtattga cacttgaggg
      361 gcgcgactac tgacagatga ggggcgcgat ccttgacact tgaggggcag agtgctgaca
      421 gatgaggggc gcacctattg acatttgagg ggctgtccac aggcagaaaa tccagcattt
      481 gcaagggttt ccgcccgttt ttcggccacc gctaacctgt cttttaacct gcttttaaac
      541 caatatttat aaaccttgtt tttaaccagg gctgcgccct gtgcgcgtga ccgcgcacgc
      601 cgaagggggg tgccccccct tctcgaaccc tcccggcccg ctaacgcggg cctcccatcc
      661 ccccaggggc tgcgcccctc ggccgcgaac ggcctcaccc caaaaatggc agccaagctg
      721 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
      781 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
      841 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
      901 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
      961 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     1021 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     1081 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     1141 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     1201 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     1261 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     1321 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     1381 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     1441 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     1501 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     1561 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     1621 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     1681 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     1741 ttttgctcac atgttcagcg cttgaacttg gcgtagcgct cgcgggcgat ttccagggtc
     1801 gagttgcgga cgcgctcgaa gcgctccttg tccttctgcc acagcgagcg gaccgccagg
     1861 ccgcggaccg agttgaagat cagccacagg atgtcctccg cgtcgtcgcg gctcaggccg
     1921 cgcgacacca gcacgcccag ccacatatcc tcgacgacga agcggttgcg ctcgacggtg
     1981 cgctggatgc cctcgcgcag cgccgggtcg cggtcggcgg ccacgatcag gtccagcgag
     2041 atcgagaagt cgtcgtccag gaagaactcg gcggcgtcgt ccagcatctg ctggatgacg
     2101 tcgtcctcgg gcttcagctt ggccaggcgg gcgcggctgc gctcggtgat ctgctcgtac
     2161 agccactcga aggtggccag cagcagttcc agcttggtcg ggaagtggtg cgactgggcg
     2221 ccgcggctga cgccggcggc gcccgggacg tcggcgatgc ggaagccggc atagcccttc
     2281 tcgcgcagga cgcccagggc ggcggcgatc agcttgccct gggtttccat cgcgcgctcg
     2341 gcctgggtgc ggcgcttcgg cgacatgatg accattcggg gtctctccct gctagcatca
     2401 gggttattgt atcatgagcg gatacatatg tcaatgtacc ggaacaaaca gacaatctgg
     2461 tctgtttgta catggttgac ggatcgccgc gaagccgcta actgcgccgg ttaacaaaca
     2521 gacaatctgg tctgtttgta actagtagag gaagcttccg catgtacccg tacgacgtcc
     2581 cggactacgc cattaatatg actctagaga caagtttgta caaaaaagct gaacgagaaa
     2641 cgtaaaatga tataaatatc aatatattaa attagatttt gcataaaaaa cagactacat
     2701 aatactgtaa aacacaacat atccagtcac tatggcggcc gcattaggca ccccaggctt
     2761 tacactttat gcttccggct cgtataatgt gtggattttg agttaggatc cggcgagatt
     2821 ttcaggagct aaggaagcta aaatggagaa aaaaatcact ggatatacca ccgttgatat
     2881 atcccaatgg catcgtaaag aacattttga ggcatttcag tcagttgctc aatgtaccta
     2941 taaccagacc gttcagctgg atattacggc ctttttaaag accgtaaaga aaaataagca
     3001 caagttttat ccggccttta ttcacattct tgcccgcctg atgaatgctc atccggaatt
     3061 ccgtatggca atgaaagacg gtgagctggt gatatgggat agtgttcacc cttgttacac
     3121 cgttttccat gagcaaactg aaacgttttc atcgctctgg agtgaatacc acgacgattt
     3181 ccggcagttt ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa acctggccta
     3241 tttccctaaa gggtttattg agaatatgtt tttcgtctca gccaatccct gggtgagttt
     3301 caccagtttt gatttaaacg tggccaatat ggacaacttc ttcgcccccg ttttcaccat
     3361 gggcaaatat tatacgcaag gcgacaaggt gctgatgccg ctggcgattc aggttcatca
     3421 tgccgtctgt gatggcttcc atgtcggcag aatgcttaat gaattacaac agtactgcga
     3481 tgagtggcag ggcggggcgt aaacgcgtcg ttccactgag cgtcagaccc cgtagaaaag
     3541 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     3601 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     3661 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     3721 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     3781 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     3841 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     3901 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     3961 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     4021 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     4081 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     4141 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     4201 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     4261 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     4321 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     4381 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatacgcgt
     4441 ggatccggct tactaaaagc cagataacag tatgcgtatt tgcgcgctga tttttgcggt
     4501 ataagaatat atactgatat gtatacccga agtatgtcaa aaagaggtgt gctatgaagc
     4561 agcgtattac agtgacagtt gacagcgaca gctatcagtt gctcaaggca tatatgatgt
     4621 caatatctcc ggtctggtaa gcacaaccat gcagaatgaa gcccgtcgtc tgcgtgccga
     4681 acgctggaaa gcggaaaatc aggaagggat ggctgaggtc gcccggttta ttgaaatgaa
     4741 cggctctttt gctgacgaga acagggactg gtgaaatgca gtttaaggtt tacacctata
     4801 aaagagagag ccgttatcgt ctgtttgtgg atgtacagag tgatattatt gacacgcccg
     4861 ggcgacggat ggtgatcccc ctggccagtg cacgtctgct gtcagataaa gtctcccgtg
     4921 aactttaccc ggtggtgcat atcggggatg aaagctggcg catgatgacc accgatatgg
     4981 ccagtgtgcc ggtctccgtt atcggggaag aagtggctga tctcagccac cgcgaaaatg
     5041 acatcaaaaa cgccattaac ctgatgttct ggggaatata aatgtcaggc tccgttatac
     5101 acagccagtc tgcaggtcga ccatagtgac tggatatgtt gtgttttaca gtattatgta
     5161 gtctgttttt tatgcaaaat ctaatttaat atattgatat ttatatcatt ttacgtttct
     5221 cgttcagctt tcttgtacaa agtggtgata tcgctagcta gaggtaccga gctccaattg
     5281 ggatccggac tacaaggacc acgacggcga ctataaggac cacgacatcg actacaagga
     5341 tgacgacgac aagtgaggat ccggtgaagt gacccgcaaa aaaaaaggcc aggggatcgc
     5401 tcccccggcc ttttttttga attcactggc cgtcgtttta caacgtcgtg actgggaaaa
     5461 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     5521 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     5581 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg
     5641 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     5701 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     5761 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     5821 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     5881 ttagcaccct ttctcggtcc ttcaacgttc ctgacaacga gcctcctttt cgccaatcca
     5941 tcgacaatca ccgcgagtcc ctgctcgaac gctgcgtccg gaccggcttc gtcgaaggcg
     6001 tctatcgcgg cccgcaacag cggcgagagc ggagcctgtt caacggtgcc gccgcgctcg
     6061 ccggcatcgc tgtcgccggc ctgctcctca agcacggccc caacagtgaa gtagctgatt
     6121 gtcatcagcg cattgacggc gtccccggcc gaaaaacccg cctcgcagag gaagcgaagc
     6181 tgcgcgtcgg ccgtttccat ctgcggtgcg cccggtcgcg tgccggcatg gatgcgcgcg
     6241 ccatcgcggt aggcgagcag cgcctgcctg aagctgcggg cattcccgat cagaaatgag
     6301 cgccagtcgt cgtcggctct cggcaccgaa tgcgtatgat tctccgccag catggcttcg
     6361 gccagtgcgt cgagcagcgc ccgcttgttc ctgaagtgcc agtaaagcgc cggctgctga
     6421 acccccaacc gttccgccag tttgcgtgtc gtcagaccgt ctacgccgac ctcgttcaac
     6481 aggtccaggg cggcacggat cactgtattc ggctgcaact ttgtcatgat tgacacttta
     6541 tcactgataa acataatatg tccaccaact tatcagtgat aaagaatccg cgcgttcaat
     6601 cggaccagcg gaggctggtc cggaggccag acgtgaaacc caacataccc ctgatcgtaa
     6661 ttctgagcac tgtcgcgctc gacgctgtcg gcatcggcct gattatgccg gtgctgccgg
     6721 gcctcctgcg cgatctggtt cactcgaacg acgtcaccgc ccactatggc attctgctgg
     6781 cgctgtatgc gttggtgcaa tttgcctgcg cacctgtgct gggcgcgctg tcggatcgtt
     6841 tcgggcggcg gccaatcttg ctcgtctcgc tggccggcgc cactgtcgac tacgccatca
     6901 tggcgacagc gcctttcctt tgggttctct atatcgggcg gatcgtggcc ggcatcaccg
     6961 gggcgactgg ggcggtagcc ggcgcttata ttgccgatat cactgatggc gatgagcgcg
     7021 cgcggcactt cggcttcatg agcgcctgtt tcgggttcgg gatggtcgcg ggacctgtgc
     7081 tcggtgggct gatgggcggt ttctcccccc acgctccgtt cttcgccgcg gcagccttga
     7141 acggcctcaa tttcctgacg ggctgtttcc ttttgccgga gtcgcacaaa ggcgaacgcc
     7201 ggccgttacg ccgggaggct ctcaacccgc tcgcttcgtt ccggtgggcc cggggcatga
     7261 ccgtcgtcgc cgccctgatg gcggtcttct tcatcatgca acttgtcgga caggtgccgg
     7321 ccgcgctttg ggtcattttc ggcgaggatc gctttcactg ggacgcgacc acgatcggca
     7381 tttcgcttgc cgcatttggc attctgcatt cactcgccca ggcaatgatc accggccctg
     7441 tagccgcccg gctcggcgaa aggcgggcac tcatgctcgg aatgattgcc gacggcacag
     7501 gctacatcct gcttgccttc gcgacacggg gatggatggc gttcccgatc atggtcctgc
     7561 ttgcttcggg tggcatcgga atgccggcgc tgcaagcaat gttgtccagg caggtggatg
     7621 aggaacgtca ggggcagctg caaggctcac tggcggcgct caccagcctg acctcgatcg
     7681 tcggacccct cctcttcacg gcgatctatg cggcttctat aacaacgtgg aacgggtggg
     7741 catggattgc aggcgctgcc ctctacttgc tctgcctgcc ggcgctgcgt cgcgggcttt
     7801 ggagcggcgc agggcaacga gccgatcgct gatcgtggaa acgataggcc tatgccatgc
     7861 gggtcaaggc gacttccggc aagctatacg cgccctagaa ttgtcaattt taatcctctg
     7921 tttatcggca gttcgtagag cgcgccgtgc gtcccgagcg atactgagcg aagcaagtgc
     7981 gtcgagcagt gcccgcttgt tcctgaaatg ccagtaaagc gctggctgct gaacccccag
     8041 ccggaactga ccccacaagg ccctagcgtt tgcaatgcac caggtcatca ttgacccagg
     8101 cgtgttccac caggccgctg cctcgcaact cttcgcaggc ttcgccgacc tgctcgcgcc
     8161 acttcttcac gcgggtggaa tccgatccgc acatgaggcg gaaggtttcc agcttgagcg
     8221 ggtacggctc ccggtgcgag ctgaaatagt cgaacatccg tcgggccgtc ggcgacagct
     8281 tgcggtactt ctcccatatg aatttcgtgt agtggtcgcc agcaaacagc acgacgattt
     8341 cctcgtcgat caggacctgg caacgggacg ttttcttgcc acggtccagg acgcggaagc
     8401 ggtgcagcag cgacaccgat tccaggtgcc caacgcggtc ggacgtgaag cccatcgccg
     8461 tcgcctgtag gcgcgacagg cattcctcgg ccttcgtgta ataccggcca ttgatcgacc
     8521 agcccaggtc ctggcaaagc tcgtagaacg tgaaggtgat cggctcgccg ataggggtgc
     8581 gcttcgcgta ctccaacacc tgctgccaca ccagttcgtc atcgtcggcc cgcagctcga
     8641 cgccggtgta ggtgatcttc acgtccttgt tgacgtggaa aatgaccttg ttttgcagcg
     8701 cctcgcgcgg gattttcttg ttgcgcgtgg tgaacagggc agagcgggcc gtgtcgtttg
     8761 gcatcgctcg catcgtgtcc ggccacggcg caatatcgaa caaggaaagc tgcatttcct
     8821 tgatctgctg cttcgtgtgt ttcagcaacg cggcctgctt ggcctcgctg acctgttttg
     8881 ccaggtcctc gccggcggtt tttcgcttct tggtcgtcat agttcctcgc gtgtcgatgg
     8941 tcatcgactt cgccaaacct gccgcctcct gttcgagacg acgcgaacgc tccacggcgg
     9001 ccgatggcgc gggcagggca gggggagcca gttgcacgct gtcgcgctcg atcttggccg
     9061 tagcttgctg gaccatcgag ccgacggact ggaaggtttc gcggggcgca cgcatgacgg
     9121 tgcggcttgc gatggtttcg gcatcctcgg cggaaaaccc cgcgtcgatc agttcttgcc
     9181 tgtatgcctt ccggtcaaac gtccgattca ttcaccctcc ttgcgggatt gccccgactc
     9241 acgccggggc aatgtgccct tattcctgat ttgacccgcc tggtgccttg gtgtccagat
     9301 aatccacctt atcggcaatg aagtcggtcc cgtagaccgt ctggccgtcc ttctcgtact
     9361 tggtattccg aatcttgccc tgcacgaata ccagctccgc gaagtcgctc ttcttgatgg
     9421 agcgcatggg gacgtgcttg gcaatcacgc gcaccccccg gccgttttag cggctaaaaa
     9481 agtcatggct ctgccctcgg gcggaccacg cccatcatga ccttgccaag ctcgtcctgc
     9541 ttctcttcga tcttcgccag cagggcgagg atcgtggcat caccgaaccg cgccgtgcgc
     9601 gggtcgtcgg tgagccagag tttcagcagg ccgcccaggc ggcccaggtc gccattgatg
     9661 cgggccagct cgcggacgtg ctcatagtcc acgacgcccg tgattttgta gccctggccg
     9721 acggccagca ggtaggccta caggctcatg ccggccgccg ccgccttttc ctcaatcgct
     9781 cttcgttcgt ctggaaggca gtacaccttg ataggtgggc tgcccttcct ggttggcttg
     9841 gtttcatcag ccatccgctt gccctcatct gttacgccgg cggtagccgg ccagcctcgc
     9901 agagcaggat tcccgttgag caccgccagg tgcgaataag ggacagtgaa gaaggaacac
     9961 ccgctcgcgg gtgggcctac ttcacctatc ctgcccggct gacgccgttg gatacaccaa
    10021 ggaaagtcta cacgaaccct ttggcaaaat cctgtatatc gtgcgaaaaa ggatggatat
    10081 accgaaaaaa tcgctataat gaccccgaag cagggttatg cagcggaaaa gatccgtc
//
