LOCUS       V005592                10513 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V005592
VERSION     V005592
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 10513)
  AUTHORS   Untergasser A, Geurts R, Franssen H.
  TITLE     pHC60 - a very stable plasmid for fluorescence mircroscopy
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 10513)
  AUTHORS   Untergasser A, Geurts R, Franssen H.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-AUG-2008) Laboratory of Molecular Biology, Wageningen
            University, Dreijenlaan 3, Wageningen 6703HA, The Netherlands
REFERENCE   3  (bases 1 to 10513)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 10513)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (19-AUG-2008) Laboratory of Molecular Biology, Wageningen
            University, Dreijenlaan 3, Wageningen 6703HA, The Netherlands"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..10513
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     508..524
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     misc_feature    525..581
                     /label="MCS"
                     /note="pUC18/19 multiple cloning site"
     promoter        613..631
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             663..685
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             693..1409
                     /label="Cycle 3 GFP"
                     /note="Cycle 3 GFP (Crameri et al., 1996)"
     terminator      1474..1521
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
     primer_bind     complement(1581..1597)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    1605..1621
                     /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(1629..1659)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1674..1695)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3178..3888)
                     /direction=LEFT
                     /label="oriV"
                     /note="incP origin of replication"
     oriT            4033..4142
                     /label="oriT"
                     /note="incP origin of transfer"
     CDS             4175..4543
                     /label="traJ"
                     /note="oriT-recognizing protein"
     CDS             7240..7488
                     /gene="parD"
                     /label="Antitoxin ParD"
                     /note="Antitoxin ParD from Escherichia coli. Accession#:
                     P22995"
     CDS             7488..7796
                     /gene="parE"
                     /label="Toxin ParE"
                     /note="Toxin ParE from Escherichia coli. Accession#:
                     Q79EC5"
     CDS             complement(8093..9172)
                     /label="lacI"
                     /note="lac repressor"
     promoter        complement(9173..9250)
                     /label="lacIq promoter"
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             complement(9312..10508)
                     /label="TcR"
                     /note="tetracycline efflux protein"
ORIGIN
        1 gcctccggac cagcctccgc tggtccgatt gaacgcgcgg attctttatc actgataagt
       61 tggtggacat attatgttta tcagtgataa agtgtcaagc atgacaaagt tgcagccgaa
      121 tacagtgatc cgtgccgccc tggacctgtt gaacgaggtc ggcgtagacg gtctgacgac
      181 acgcaaactg gcggaacggt tgggggttca gcagccggcg ctttactggc acttcaggaa
      241 caagcgggcg ctgctcgacg cactggccga agccatgctg gcggagaatc atacgcattc
      301 ggtgccgaga gccgacgacg actggcgctc atttctgatc gggaatgccc gcagcttcag
      361 gcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
      421 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
      481 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaattc gagctcggta
      541 cccggggatc ctctagagtc gacctgcagg catgcaagct ttcgcgagct cgagatctcg
      601 atcccgcgaa attaatacga ctcactatag ggagaccaca acggtttccc tctagaaata
      661 attttgttta actttaagaa ggagatatac atatggctag caaaggagaa gaactcttca
      721 ctggagttgt cccaattctt gttgaattag atggtgatgt taatgggcac aaattttctg
      781 tcagtggaga gggtgaaggt gatgcaacat acggaaaact tacccttaaa tttatttgca
      841 ctactggaaa actacctgtt ccatggccaa cacttgtcac tactttcact tatggtgttc
      901 agtgcttttc aagatacccg gatcatatga aacggcatga ctttttcaag agtgccatgc
      961 ccgaaggtta tgtacaggaa agaactatat ttttcaaaga tgacgggaac tacaagacac
     1021 gtgctgaagt caagtttgaa ggtgataccc ttgttaatag aatcgagtta aaaggtattg
     1081 attttaaaga agatggaaac attcttgggc acaaattgga atacaactat aactcacaca
     1141 atgtatacat catggcagac aaacaaaaga atggaatcaa agctaacttc aaaattagac
     1201 acaacattga agatggaagc gttcaactag cagaccatta tcaacaaaat actccaattg
     1261 gcgatggccc tgtcctttta ccagacaacc attacctgtc cacacaatct gccctttcga
     1321 aagatcccaa cgaaaagaga gaccacatgg tccttcttga gtttgtaaca gctgctggga
     1381 ttacacatgg catggatgaa ctatacaaag gatccggctg ctaacaaagc ccgaaaggaa
     1441 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
     1501 cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggattcga gatctagata
     1561 tcgatgaatt cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     1621 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     1681 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     1741 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     1801 tgctcgccta ccgcgatggc gcgcgcatcc atgccggcac gcgaccgggc gcaccgcaga
     1861 tggaaacggc cgacgcgcag cttcgcttcc tctgcgaggc gggtttttcg gccggggacg
     1921 ccgtcaatgc gctgatgaca atcagctact tcactgttgg ggccgtgctt gaggagcagg
     1981 ccggcgacag cgatgccggc gagcgcggcg gcaccgttga acaggctccg ctctcgccgc
     2041 tgttgcgggc cgcgatagac gccttcgacg aagccggtcc ggacgcagcg ttcgagcagg
     2101 gactcgcggt gattgtcgat ggattggcga aaaggaggct cgttgtcagg aacgttgaag
     2161 gaccgagaaa gggtgacgat tgatcaggac cgctgccgga gcgcaaccca ctcactacag
     2221 cagagccatg tagacaacat cccctccccc tttccaccgc gtcagacgcc cgtagcagcc
     2281 cgctacgggc tttttcatgc cctgccctag cgtccaagcc tcacggccgc gctcggcctc
     2341 tctggcggcc ttctggcgct cctgctgcgg cgtccgctcg tgggccgtgg cgcgggtccg
     2401 cgcgccggcc tcgtgcgcct ggcgctcgcg ggcgaggtcc agggcggccg tcttcacgtt
     2461 ctgccttgcg cagatgagat agatccagtc tgcaagtgtc agaaacgagt ctgcatttga
     2521 tgccctgaac gtcttcatac atctgataag cccctcccgg aagtctgctt tctctccgtc
     2581 agaaagcata ccctgaccgg tttcaatcac ggaccgttct gtcaggtaat gctctgtcca
     2641 gtcatacaga gccgggtttg tcacccgtat aagctgaagc aggcacaaat cagggaaata
     2701 aacaaaatcc cgcatccccg gataaagaaa aatcagggaa ttaatggcct gatggatttc
     2761 ccgtggcgtc gaaagtgcgg caccataggt atcagtcacc gcgatgagat cccttaccat
     2821 tccagagtct ggcggttgat tattaatttg ctgatataga gcctcagccc gctggcgaaa
     2881 ttcattacgt aaatcaaagg cttcaggtcg gggtaattta aaactaagct gaatgatttt
     2941 ctggagataa cggctgccat cttcgatatt cagcgcatgt tcaacggcat gagtgataat
     3001 ctgcctgtca taacagagaa tatgggtaaa gcggggcaga tcggctactg cacgcacaag
     3061 cctgaacact tccgccacct gggatggctc cagtcggtcc agatcatcca tgacaacaat
     3121 gaacttcaga tccagactca ccagttgtcc tgcaatttca gcccgaagct agcttggctg
     3181 ccatttttgg ggtgaggccg ttcgcggccg aggggcgcag cccctggggg gatgggaggc
     3241 ccgcgttagc gggccgggag ggttcgagaa gggggggcac cccccttcgg cgtgcgcggt
     3301 cacgcgcaca gggcgcagcc ctggttaaaa acaaggttta taaatattgg tttaaaagca
     3361 ggttaaaaga caggttagcg gtggccgaaa aacgggcgga aacccttgca aatgctggat
     3421 tttctgcctg tggacagccc ctcaaatgtc aataggtgcg cccctcatct ggtcagcact
     3481 ctgcccctca agtgtcaagg atcgcgcccc tcatctgtca gtagtcgcgc ccctcaagtg
     3541 tcaataccgc agggcactta tccccaggct tgtccacatc atctgtggga aactcgcgta
     3601 aaatcaggcg ttttcgccga tttgcgaggc tggccagctc cacgtcgccg gccgaaatcg
     3661 agcctgcccc tcatctgtca acgccgcgcc gggtgagtcg gcccctcaag tgtcaacgtc
     3721 cgcccctcat ctgtcagtga gggccaagtt ttccgcgagg tatccacaac gccggcggcc
     3781 gcggtgtctc gcacacggct tcgacggcgt ttctggcgcg tttgcagggc catagacggc
     3841 cgccagccca gcggcgaggg caaccagccc ggtgagcgtc ggaaagggtc gacggatctt
     3901 ttccgctgca taaccctgct tcggggtcat tatagcgatt ttttcggtat atccatcctt
     3961 tttcgcacga tatacaggat tttgccaaag ggttcgtgta gactttcctt ggtgtatcca
     4021 acggcgtcag ccgggcagga taggtgaagt aggcccaccc gcgagcgggt gttccttctt
     4081 cactgtccct tattcgcacc tggcggtgct caacgggaat cctgctctgc gaggctggcc
     4141 ggctaccgcc ggcgtaacag atgagggcaa gcggatggct gatgaaacca agccaaccag
     4201 gaagggcagc ccacctatca aggtgtactg ccttccagac gaacgaagag cgattgagga
     4261 aaaggcggcg gcggccggca tgagcctgtc ggcctacctg ctggccgtcg gccagggcta
     4321 caaaatcacg ggcgtcgtgg actatgagca cgtccgcgag ctggcccgca tcaatggcga
     4381 cctgggccgc ctgggcggcc tgctgaaact ctggctcacc gacgacccgc gcacggcgcg
     4441 gttcggtgat gccacgatcc tcgccctgct ggcgaagatc gaagagaagc aggacgagct
     4501 tggcaaggtc atgatgggcg tggtccgccc gagggcagag ccatgacttt tttagccgct
     4561 aaaacggccg gggggtgcgc gtgattgcca agcacgtccc catgcgctcc atcaagaaga
     4621 gcgacttcgc ggagctggta ttcgtgcagg gcaagattcg gaataccaag tacgagaagg
     4681 acggccagac ggtctacggg accgacttca ttgccgataa ggtggattat ctggacacca
     4741 aggcaccagg cgggtcaaat caggaataag ggcacattgc cccggcgtga gtcggggcaa
     4801 tcccgcaagg agggtgaatg aatcggacgt ttgaccggaa ggcatacagg caagaactga
     4861 tcgacgcggg gttttccgcc ggatgatgcc gaaaccatcg caagccgcac cgtcatgcgt
     4921 gcgccccgcg aaaccttcca gtccgtcggc tcgatggtcc agcaagctac ggccaagatc
     4981 gagcgcgaca gcgtgcaact ggctcccctg ccctgcccgc gccatcggcc gccgtggagc
     5041 gttcgcgtcg tctcgaacag gaggcggcag gtttggcgaa gtcgatgacc atcgacacgc
     5101 gaggaactat gacgaccaag aagcgaaaaa ccgccggcga ggacctggca aaacaggtca
     5161 gcgaggccaa gcaggccgcg ttgctgaaac acacgaagca gcagatcaag gaaatgcagc
     5221 tttccttgtt cgatattgcg ccgtggccgg acacgatgcg agcgatgcca aacgacacgg
     5281 cccgctctgc cctgttcacc acgcgcaaca agaaaatccc gcgcgaggcg ctgcaaaaca
     5341 aggtcatttt ccacgtcaac aaggacgtga agatcaccta caccggcgtc gagctgcggg
     5401 ccgacgatga cgaactggtg tggcagcagg tgttggagta cgcgaagcgc acccctatcg
     5461 gcgagccgat caccttcacg ttctacgagc tttgccagga cctgggctgg tcgatcaatg
     5521 gccggtatta cacgaaggcc gaggaatgcc tgtcgcgcct acagggcgac ggcgatgggc
     5581 ttcacgtccg accgcgttgg gcacctggaa tcggtgtcgc tgctgcaccg cttccgcgtc
     5641 ctggaccgtg gcaagaaaac gtcccgttgc caggtcctga tcgacgagga aatcgtcgtg
     5701 ctgtttgctg gcgaccacta cacgaaattc atatgggaga agtaccgcaa gctgtcgccg
     5761 acggcccgac ggatgttcga ctatttcagc tcgcaccggg agccgtaccc gctcaagctg
     5821 gaaaccttcc gcctcatgtg cggatcggat tccacccgcg tgaagaagtg gcgcgagcag
     5881 gtcggcgaag cntgcgaaga gttgcgaggc agcggcctgg tggaacacgc ctgggtcaat
     5941 gatgacctgg tgcattgcaa acgctagggc cttgtggggt cagttccggc tgggggttca
     6001 gcagccagcg ctttactggc atttcaggaa caagcgggca ctgctcgacg cacttgcttc
     6061 gctcagtatc gctcgggacg cacggcgcgc tctacgaact gccgataaac agaggattaa
     6121 aattgacaat tgtgattaag gctcagattc gacggcttgg agcggccgac gtgcaggatt
     6181 tccgcgagat ccgattgtcg gccctgaaga aagctccaga gatgttcggg tccgtttacg
     6241 agcacgagga gaaaaagccc atggaggcgt tcgctgaacg gttgcgagat gccgtggcat
     6301 tcggcgccta catcgacggc gagatcattg ggctgtcggt cttcaaacag gaggacggcc
     6361 ccaaggacgc tcacaaggcg catctgtccg gcgttttcgt ggagcccgaa cagcgaggcc
     6421 gaggggtcgc cggtatgctg ctgcgggcgt tgccggcggg tttattgctc gtgatgatcg
     6481 tccgacagat tcccaacggg aatctggtgg atgcgcatct tcatcctcgg cgcacttaat
     6541 atttcgctat tctggagctt gttgtttatt tcggtctacc gcctgccggg cggggtcgcg
     6601 gcgacggtag gcgctgtgca gccgctgatg gtcgtgttca tctctgccgc tctgctaggt
     6661 agcccgatac gattgatggc ggtcctgggg gctatttgcg gaactgcggg cgtggcgctg
     6721 ttggtgttga caccaaacgc agcgctagat cctgtcggcg tcgcagcggg cctggcgggg
     6781 gcggtttcca tggcgttcgg aaccgtgctg acccgcaagt ggcaacctcc cgtgcctctg
     6841 ctcaccttta ccgcctggca actggcggcc ggaggacttc tgctcgttcc agtagcttta
     6901 gtgtttgatc cgccaatccc gatgcctaca ggaaccaatg ttctcggcct ggcgtggctc
     6961 ggcctgatcg gagcgggttt aacctacttc ctttggttcc gggggatctc gcgactcgaa
     7021 cctacagttg tttccttact gggctttctc agccccggcg tcccccttgg tcaaattggg
     7081 tatacccatt tgggcctagt ctagccggca tggcgcatta cagcaatacg caatttaaat
     7141 gcgcctagcg cattttcccg accttaatgc gcctcgcgct gtagcctcac gcccacatat
     7201 gtgctaatgt ggttacgtgt attttatgga ggttatccaa tgagccgcct gacaatcgac
     7261 atgacggacc agcagcacca gagcctgaaa gccctggccg ccttgcaggg caagaccatt
     7321 aagcaatacg ccctcgaacg tctgttcccc ggtgacgctg atgccgatca ggcatggcag
     7381 gaactgaaaa ccatgctggg gaaccgcatc aacgatgggc ttgccggcaa ggtgtccacc
     7441 aagagcgtcg gcgaaattct tgatgaagaa ctcagcgggg atcgcgcttg acggcctaca
     7501 tcctcacggc tgaggccgaa gccgatctac gcggcatcat ccgctacacg cgccgggagt
     7561 ggggcgcggc gcaggtgcgc cgctatatcg ctaagctgga acagggcata gccaggcttg
     7621 ccgccggcga aggcccgttt aaggacatga gcgaactctt tcccgcgctg cggatggccc
     7681 gctgcgaaca ccactacgtt ttttgcctgc cgcgtgcggg cgaacccgcg ttggtcgtgg
     7741 cgatcctgca tgagcgcatg gacctcatga cgcgacttgc cgacaggctc aagggctgat
     7801 ttcagccgct aaaaatcgcg ccactcacaa cgtcctgatg gcgtactcaa gcttgatatc
     7861 gaattcctgc agcccggggg atcggggacc gccgtgttgc taggatggtt gttcttggat
     7921 cagacgctga gtgcgcttca aatcatcggc gtcctgctcg tgatcgggag tatctggctg
     7981 ggccaacgtt ccaaccgcac tcctagggcg cgtatagctt gccggaagtc gccttgaccc
     8041 gcatggcata ggttgcaatt cgcgctaact tacattaatt gcgttgcgct cactgcccgc
     8101 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     8161 aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca
     8221 gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt
     8281 gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa catgagctgt
     8341 cttcggtatc gtcgtatccc actaccgaga tatccgcacc aacgcgcagc ccggactcgg
     8401 taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa
     8461 cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc
     8521 cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca
     8581 gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac
     8641 ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac
     8701 tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag
     8761 cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc
     8821 gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca
     8881 tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt
     8941 gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc
     9001 ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca
     9061 ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct
     9121 gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca
     9181 ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcaccatt
     9241 cgatggtgtc aacgtaaatg catgccgctt cgccttcgcg cgcgaattgc aacctatcgt
     9301 ttccacgatc agcgatcggc tcgttgccct gcgccgctcc aaagcccgcg acgcagcgcc
     9361 ggcaggcaga gcaagtagag ggcagcgcct gcaatccatg cccacccgtt ccacgttgtt
     9421 atagaagccg catagatcgc cgtgaagagg aggggtccga cgatcgaggt caggctggtg
     9481 agcgccgcca gtgagccttg cagctgcccc tgacgttcct catccacctg cctggacaac
     9541 attgcttgca gcgccggcat tccgatgcca cccgaagcaa gcaggaccat gatcgggaac
     9601 gccatccatc cccgtgtcgc gaaggcaagc aggatgtagc ctgtgccgtc ggcaatcatt
     9661 ccgagcatga gtgcccgcct ttcgccgagc cgggcggcta cagggccggt gatcattgcc
     9721 tgggcgagtg aatgcagaat gccaaatgcg gcaagcgaaa tgccgatcgt ggtcgcgtcc
     9781 cagtgaaagc gatcctcgcc gaaaatgacc caaagcgcgg ccggcacctg tccgacaagt
     9841 tgcatgatga agaagaccgc catcagggcg gcgacgacgg tcatgccccg ggcccaccgg
     9901 aacgaagcga gcgggttgag agcctcccgg cgtaacggcc ggcgttcgcc tttgtgcgac
     9961 tccggcaaaa ggaaacagcc cgtcaggaaa ttgaggccgt tcaaggctgc cgcggcgaag
    10021 aacggagcgt ggggggagaa accgcccatc agcccaccga gcacaggtcc cgcgaccatc
    10081 ccgaacccga aacaggcgct catgaagccg aagtgccgcg cgcgctcatc gccatcagtg
    10141 atatcggcaa tataagcgcc ggctaccgcc ccagtcgccc cggtgatgcc ggccacgatc
    10201 cgcccgatat agagaaccca aaggaaaggc gctgtcgcca tgatggcgta gtcgacagtg
    10261 gcgccggcca gcgagacgag caagattggc cgccgcccga aacgatccga cagcgcgccc
    10321 agcacaggtg cgcaggcaaa ttgcaccaac gcatacagcg ccagcagaat gccatagtgg
    10381 gcggtgacgt cgttcgagtg aaccagatcg cgcaggaggc ccggcagcac cggcataatc
    10441 aggccgatgc cgacagcgtc gagcgcgaca gtgctcagaa ttacgatcag gggtatgttg
    10501 ggtttcacgt ctg
//
