LOCUS       V005304                 7876 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V005304
VERSION     V005304
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7876)
  AUTHORS   Trachtmann N, Alvarez Fong KF, Guitart Font E, Sprenger GA.
  TITLE     Construction of chromosomally encoded lacZ and gfp reporter strains
            of Escherichia coli for global regulation of metabolism
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7876)
  AUTHORS   Trachtmann N, Alvarez Fong KF, Guitart Font E, Sprenger GA.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-MAY-2016) Institute of microbiology, Stuttgart
            university, Allmandring 31, Stuttgart 70569, Germany
REFERENCE   3  (bases 1 to 7876)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7876)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (24-MAY-2016) Institute of microbiology, Stuttgart university,
            Allmandring 31, Stuttgart 70569, Germany"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7876
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    6..303
                     /label="flanking arm for integration"
                     /note="flanking arm for integration"
     regulatory      400..520
                     /label="5S terminator"
                     /note="5S terminator"
                     /regulatory_class="terminator"
     terminator      522..608
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      700..727
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        746..837
                     /label="AmpR promoter"
     CDS             838..1695
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      1869..2457
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     misc_feature    complement(2643..2783)
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     CDS             complement(2888..3076)
                     /label="rop"
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     CDS             complement(4097..5176)
                     /label="lacI"
                     /note="lac repressor"
     promoter        complement(5177..5254)
                     /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             5605..5865
                     /gene="rpsT"
                     /label="Small ribosomal subunit protein bS20"
                     /note="Small ribosomal subunit protein bS20 from
                     Escherichia coli (strain K12). Accession#: P0A7U7"
     protein_bind    complement(5912..5959)
                     /label="FRT"
                     /bound_moiety="FLP recombinase from the Saccharomyces
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(5995..6633)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /function="chloramphenicol resistance"
                     /label="cat"
                     /protein_id="APZ86792.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRCLMNTTVLR"
     gene            complement(5995..6633)
                     /gene="cat"
                     /label="cat"
     promoter        complement(6634..6736)
                     /label="cat promoter"
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     protein_bind    complement(6842..6875)
                     /label="FRT (minimal)"
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     promoter        7067..7095
                     /label="tac promoter"
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    7103..7119
                     /label="lac operator"
                     /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)."
     CDS             7159..7872
                     /label="yeGFP"
                     /note="yeast-enhanced green fluorescent protein"
ORIGIN
        1 gatcctcaag cgggcaaata atgagccgag tcaaaatcgt ggtaatgtac aactaatcta
       61 tctcccccac cttaactttg ggggagctgc cgattaacgc accgctggac taaaaagcgc
      121 agaataatcc gtattgcaga ttcgctgtac cgccgggtgc tgaatcatcc gctcagcaaa
      181 aatagcatgg tactcttcca tcacattctc gacgcgacca atttctacga cagttttatc
      241 ggcataaaag tcatatgcat aaagcgttgg ggcaacgaag attgcattgt gcatcgcacc
      301 aaaggtaccg agctcgaatt cctgttttgg cggatgagag aagattttca gcctgataca
      361 gattaaatca gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc
      421 ggtggtccca cctgacccca tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag
      481 tgtggggtct ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc
      541 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta
      601 ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca acggcccgga gggtggcggg
      661 caggacgccc gccataaact gccaggcatc aaattaagca gaaggccatc ctgacggatg
      721 gcctttttgc gtttctacaa actcttttgt ttatttttct aaatacattc aaatatgtat
      781 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
      841 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
      901 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
      961 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     1021 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     1081 gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     1141 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     1201 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     1261 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     1321 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     1381 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     1441 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     1501 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     1561 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     1621 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     1681 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     1741 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     1801 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     1861 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     1921 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     1981 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     2041 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     2101 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     2161 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     2221 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     2281 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     2341 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     2401 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     2461 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     2521 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     2581 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     2641 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc
     2701 actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc
     2761 tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac
     2821 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     2881 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat
     2941 cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga
     3001 gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt
     3061 tttcctgttt ggtcacttga tgcctccgtg taagggggaa tttctgttca tgggggtaat
     3121 gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg
     3181 gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa
     3241 aatcactcag ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag
     3301 ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt
     3361 ttccagactt tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga
     3421 cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc
     3481 agtaaggcaa ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac
     3541 ccgtggccag gacccaacgc tgcccgagat gcgccgcgtg cggctgctgg agatggcgga
     3601 cgcgatggat atgttctgcc aagggttggt ttgcgcattc acagttctcc gcaagaattg
     3661 attggctcca attcttggag tggtgaatcc gttagcgagg tgccgccggc ttccattcag
     3721 gtcgaggtgg cccggctcca tgcaccgcga cgcaacgcgg ggaggcagac aaggtatagg
     3781 gcggcgccta caatccatgc caacccgttc catgtgctcg ccgaggcggc ataaatcgcc
     3841 gtgacgatca gcggtccagt gatcgaagtt aggctggtaa gagccgcgag cgatccttga
     3901 agctgtccct gatggtcgtc atctacctgc ctggacagca tggcctgcaa cgcgggcatc
     3961 ccgatgccgc cggaagcgag aagaatcata atggggaagg ccatccagcc tcgcgtcgcg
     4021 aacgccagca agacgtagcc cagcgcgtcg gccagcttgc aattcgcgct aacttacatt
     4081 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     4141 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc
     4201 ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt
     4261 gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta
     4321 acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatatccg
     4381 caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt
     4441 tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa
     4501 aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag
     4561 tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg
     4621 ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac
     4681 cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata
     4741 acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat
     4801 agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg
     4861 cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc
     4921 gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa
     4981 cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat
     5041 tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct
     5101 ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata
     5161 acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca
     5221 taccgcgaaa ggttttgcac cattcgatgg tgtcaacgta aatgccgctt cgccttcgcg
     5281 cgcgaattgc aagctgatcc gggcttatcg actgcacggt gcaccaatgc ttctggcgtc
     5341 aggcagccat cgagatccaa agaagtggct tttcattgcc atggcgcaaa tcccgggaag
     5401 aaactgaccg cctgctgcaa tttttatcgc ggaaaagctg tattcacacc ccgcaagctg
     5461 gtagaatcct gcgccatcac tacgtaacga gtgccggcac attaacggcg cttatttgca
     5521 caaatccatt gacaaaagaa ggctaaaagg gcatattcct cggcctttga attgtccata
     5581 tagaacacat ttgggagttg gaccttggct aatatcaaat cagctaagaa gcgcgccatt
     5641 cagtctgaaa aggctcgtaa gcacaacgca agccgtcgct ctatgatgcg tactttcatc
     5701 aagaaagtat acgcagctat cgaagctggc gacaaagctg ctgcacagaa agcatttaac
     5761 gaaatgcaac cgatcgtgga ccgtcaggct gctaaaggtc tgatccacaa aaacaaagct
     5821 gcacgtcata aggctaacct gactgcacag atcaacaaac tggcttaatc gccaatttgc
     5881 tgagatccat tgtgtaggct ggagctgctt cgaagttcct atactttcta gagaatagga
     5941 acttcggaat aggaacttca tttaaatggc gcgccttacg ccccgccctg ccactcatcg
     6001 cagtactgtt gtattcatta agcatctgcc gacatggaag ccatcacaaa cggcatgatg
     6061 aacctgaatc gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatggt
     6121 gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac tggtgaaact
     6181 cacccaggga ttggctgaga cgaaaaacat attctcaata aaccctttag ggaaataggc
     6241 caggttttca ccgtaacacg ccacatcttg cgaatatatg tgtagaaact gccggaaatc
     6301 gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta
     6361 acaagggtga acactatccc atatcaccag ctcaccgtct ttcattgcca tacgtaattc
     6421 cggatgagca ttcatcaggc gggcaagaat gtgaataaag gccggataaa acttgtgctt
     6481 atttttcttt acggtcttta aaaaggccgt aatatccagc tgaacggtct ggttataggt
     6541 acattgagca actgactgaa atgcctcaaa atgttcttta cgatgccatt gggatatatc
     6601 aacggtggta tatccagtga tttttttctc cattttagct tccttagctc ctgaaaatct
     6661 cgacaactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa agttggaacc
     6721 tcttacgtgc cgatcaacgt ctcattttcg ccaaaagttg gcccagggct tcccggtatc
     6781 aacagggaca ccaggattta tttattctgc gaagtgatct tccgtcacag gtaggcgcgc
     6841 cgaagttcct atactttcta gagaatagga acttcggaat aggaactaag gaggatattc
     6901 atatggacca tggctaattc ccatgtcagc cgttaagatc tgtggtatgg ctgtgcaggt
     6961 cgtaaatcac tgcataattc gtgtcgctca aggcgcactc ccgttctgga taatgttttt
     7021 tgcgccgaca tcataacggt tctggcaaat attctgaaat gagctgttga caattaatca
     7081 tcggctcgta taatgtgtgg aattgtgagc ggataacaat ttcacacagg aaacaaagct
     7141 taagaaggag atatacatat gagtaaagga gaagaacttt tcactggagt tgtcccaatt
     7201 cttgttgaat tagatggtga tgttaatggg cacaaatttt ctgtcagtgg agagggtgaa
     7261 ggtgatgcaa catacggaaa acttaccctt aaatttattt gcactactgg aaaactacct
     7321 gttccatggc caacacttgt cactactttc gcgtatggtc ttcaatgctt tgcgagatac
     7381 ccagatcata tgaaacagca tgactttttc aagagtgcca tgcccgaagg ttatgtacag
     7441 gaaagaacta tatttttcaa agatgacggg aactacaaga cacgtgctga agtcaagttt
     7501 gaaggtgata cccttgttaa tagaatcgag ttaaaaggta ttgattttaa agaagatgga
     7561 aacattcttg gacacaaatt ggaatacaac tataactcac acaatgtata catcatggca
     7621 gacaaacaaa agaatggaat caaagttaac ttcaaaatta gacacaacat tgaagatgga
     7681 agcgttcaac tagcagacca ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt
     7741 ttaccagaca accattacct gtccacacaa tctgcccttt cgaaagatcc caacgaaaag
     7801 agagaccaca tggtccttct tgagtttgta acagctgctg ggattacaca tggcatggat
     7861 gaactataca aataag
//
