LOCUS       40924_25311        9499 bp DNA     circular SYN 18-DEC-2018
DEFINITION  TREX vector pIC20H-RL, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9499)
  AUTHORS   Loeschcke A, Markert A, Wilhelm S, Wirtz A, Rosenau F, Jaeger KE, 
            Drepper T.
  TITLE     TREX: A Universal Tool for the Transfer and Expression of 
            Biosynthetic Pathways in Bacteria
  JOURNAL   ACS Synth Biol 2 (1), 22-33 (2013)
  PUBMED    23656323
REFERENCE   2  (bases 1 to 9499)
  AUTHORS   Loeschcke A, Markert A, Drepper T, Jaeger K-E.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-SEP-2012) Institute of Molecular Enzyme Technology, 
            Heinrich-Heine-University, Stetternicher Forst, Juelich, NRW 52426, 
            Germany
REFERENCE   3  (bases 1 to 9499)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9499)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth 
            Biol"; date: "2013"; volume: "2"; issue: "1"; pages: "22-33"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (11-SEP-2012) Institute of Molecular Enzyme Technology, 
            Heinrich-Heine-University, Stetternicher Forst, Juelich, NRW 52426, 
            Germany"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..9499
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    106..127
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        142..172
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /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)."
     primer_bind     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(274..292)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             326..1117
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     misc_feature    1178..1196
                     /note="OE; outside end of transposon Tn5"
     oriT            1247..2438
                     /label=origin of conjuational transfer
                     /note="origin of conjuational transfer"
     promoter        2537..2565
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     CDS             2613..3800
                     /label=TcR
                     /note="tetracycline efflux protein"
     misc_feature    complement(3861..3879)
                     /note="OE; outside end of transposon Tn5"
     CDS             3953..5380
                     /label=Tn5 transposase
                     /note="transposase from the bacterial Tn5 transposon
                     (Reznikoff, 1993)"
     CDS             complement(5486..6016)
                     /label=GmR
                     /note="gentamycin acetyltransferase"
     promoter        complement(6205..6233)
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             complement(6299..7015)
                     /label=EYFP
                     /note="enhanced YFP"
     promoter        7027..7045
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    7046..7070
                     /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)."
     primer_bind     complement(7103..7119)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7593..7697
                     /label=AmpR promoter
     CDS             7698..8555
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      8729..9317
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc
      241 atgcctgcag gtcgactcta gaggatcccc actcctatag tgagtcgtat taattgatca
      301 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
      361 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
      421 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
      481 cctgtccggt gccctgaatg aactccaaga cgaggcagcg cggctatcgt ggctggccac
      541 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
      601 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
      661 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
      721 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
      781 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
      841 caggctcaag gcgcggatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
      901 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
      961 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     1021 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     1081 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     1141 atgacagatc ctcgctacct taggaccgtt atagttaact tgtgtataag agtcagagat
     1201 cagggccgag cgaagcgagc cagccggtgg ccgctcgcgg ccatcgtcca catatccacg
     1261 ggctggcaag ggagcgcagc gaccgcgcag ggcgaagccc ggagagcaag cccgtagggc
     1321 gccgcagccg ccgtaggcgg tcacgacttt gcgaagcaaa gtctagtgag tatactcaag
     1381 cattgagtgg cccgccggag gcaccgcctt gcgctgcccc cgtcgagccg gttggacacc
     1441 aaaagggagg ggcaggcatg gcggcatacg cgatcatgcg atgcaagaag ctggcgaaaa
     1501 tgggcaacgt ggcggccagt ctcaagcacg cctaccgcga gcgcgagacg cccaacgctg
     1561 acgccagcag gacgccagag aacgagcact gggcggccag cagcaccgat gaagcgatgg
     1621 gccgactgcg cgagttgctg ccagagaagc ggcgcaagga cgctgtgttg gcggtcgagt
     1681 acgtcatgac ggccagcccg gaatggtgga agtcggccag ccaagaacag caggcggcgt
     1741 tcttcgagaa ggcgcacaag tggctggcgg acaagtacgg ggcggatcgc atcgtgacgg
     1801 ccagcatcca ccgtgacgaa accagcccgc acatgaccgc gttcgtggtg ccgctgacgc
     1861 aggacggcag gctgtcggcc aaggagttca tcggcaacaa agcgcagatg acccgcgacc
     1921 agaccacgtt tgcggccgct gtggccgatc tagggctgca acggggcatc gagggcagca
     1981 aggcacgtca cacgcgcatt caggcgttct acgaggccct ggagcggcca ccagtgggcc
     2041 acgtcaccat cagcccgcaa gcggtcgagc cacgcgccta tgcaccgcag ggattggccg
     2101 aaaagctggg aatctcaaag cgcgttgaga cgccggaagc cgtggccgac cggctgacaa
     2161 aagcggttcg gcaggggtat gagcctgccc tacaggccgc cgcaggagcg cgtgagatgc
     2221 gcaagaaggc cgatcaagcc caagagacgg cccgagacct tcgggagcgc ctgaagcccg
     2281 ttctggacgc cctggggccg ttgaatcggg atatgcaggc caaggccgcc gcgatcatca
     2341 aggccgtggg cgaaaagctg ctgacggaac agcgggaagt ccagcgccag aaacaggccc
     2401 agcgccagca ggaacgcggg cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     2461 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     2521 tcaagaattc tcatgtttga cagcttatca tcgataagct ttaatgcggt agtttatcac
     2581 agttaaattg ctaacgcagt caggcaccgt gtatgaaatc taacaatgcg ctcatcgtca
     2641 tcctcggcac cgtcaccctg gatgctgtag gcataggctt ggttatgccg gtactgccgg
     2701 gcctcttgcg ggatatcgtc cattccgaca gcatcgccag tcactatggc gtgctgctag
     2761 cgctatatgc gttgatgcaa tttctatgcg cacccgttct cggagcactg tccgaccgct
     2821 ttggccgccg cccagtcctg ctcgcttcgc tacttggagc cactatcgac tacgcgatca
     2881 tggcgaccac acccgtcctg tggatcctct acgccggacg catcgtggcc ggcatcaccg
     2941 gcgccacagg tgcggttgct ggcgcctata tcgccgacat caccgatggg gaagatcggg
     3001 ctcgccactt cgggctcatg agcgcttgtt tcggcgtggg tatggtggca ggccccgtgg
     3061 ccgggggact gttgggcgcc atctccttgc atgcaccatt ccttgcggcg gcggtgctca
     3121 acggcctcaa cctactactg ggctgcttcc taatgcagga gtcgcataag ggagagcgtc
     3181 gaccgatgcc cttgagagcc ttcaacccag tcagctcctt ccggtgggcg cggggcatga
     3241 ctatcgtcgc cgcacttatg actgtcttct ttatcatgca actcgtagga caggtgccgg
     3301 cagcgctctg ggtcattttc ggcgaggacc gctttcgctg gagcgcgacg atgatcggcc
     3361 tgtcgcttgc ggtattcgga atcttgcacg ccctcgctca agccttcgtc actggtcccg
     3421 ccaccaaacg tttcggcgag aagcaggcca ttatcgccgg catggcggcc gacgcgctgg
     3481 gctacgtctt gctggcgttc gcgacgcgag gctggatggc cttccccatt atgattcttc
     3541 tcgcttccgg cggcatcggg atgcccgcgt tgcaggccat gctgtccagg caggtagatg
     3601 acgaccatca gggacagctt caaggatcgc tcgcggctct taccagccta acttcgatca
     3661 ttggaccgct gatcgtcacg gcgatttatg ccgcctcggc gagcacatgg aacgggttgg
     3721 catggattgt aggcgccgcc ctataccttg tctgcctccc cgcgttgcgt cgcggtgcat
     3781 ggagccgggc cacctcgacc tgaatggaag ccggcggcac ctcgctaaca gtgggtaccg
     3841 agctcgaatt catcgatggg ctgactctta tacacaagta gcgtcctgaa cggaaccttt
     3901 cccgttttcc aggatctgat cttccatgtg acctcctaac atggtaacgt tcatgataac
     3961 ttctgctctt catcgtgcgg ccgactgggc taaatctgtg ttctcttcgg cggcgctggg
     4021 tgatcctcgc cgtactgccc gcttggttaa cgtcgccgcc caattggcaa aatattctgg
     4081 taaatcaata accatctcat cagagggtag tgaagccatg caggaaggcg cttaccgatt
     4141 tatccgcaat cccaacgttt ctgccgaggc gatcagaaag gctggcgcca tgcaaacagt
     4201 caagttggct caggagtttc ccgaactgct ggccattgag gacaccacct ctttgagtta
     4261 tcgccaccag gtcgccgaag agcttggcaa gctgggctct attcaggata aatcccgcgg
     4321 atggtgggtt cactccgttc tcttgctcga ggccaccaca ttccgcaccg taggattact
     4381 gcatcaggag tggtggatgc gcccggatga ccctgccgat gcggatgaaa aggagagtgg
     4441 caaatggctg gcagcggccg caactagccg gttacgcatg ggcagcatga tgagcaacgt
     4501 gattgcggtc tgtgaccgcg aagccgatat tcatgcttat ctgcaggaca aactggcgca
     4561 taacgagcgc ttcgtggtgc gctccaagca cccacgcaag gacgtagagt ctgggttgta
     4621 tctgtacgac catctgaaga accaaccgga gttgggtggc tatcagatca gcattccgca
     4681 aaagggcgtg gtggataaac gcggtaaacg taaaaatcga ccagcccgca aggcgagctt
     4741 gagcctgcgc agtgggcgca tcacgctaaa acaggggaat atcacgctca acgcggtgct
     4801 ggccgaggag attaacccgc ccaagggtga gaccccgttg aaatggttgt tgctgaccag
     4861 cgaaccggtc gagtcgctag cccaagcctt gcgcgtcatc gacatttata cccatcgctg
     4921 gcggatcgag gagttccata aggcatggaa aaccggagca ggagccgaga ggcaacgcat
     4981 ggaggagccg gataatctgg agcggatggt ctcgatcctc tcgtttgttg cggtcaggct
     5041 gttacagctc agagaaagct tcacgctgcc gcaagcactc agggcgcaag ggctgctaaa
     5101 ggaagcggaa cacgtagaaa gccagtccgc agaaacggtg ctgaccccgg atgaatgtca
     5161 gctactgggc tatctggaca agggaaaacg caagcgcaaa gagaaagcag gtagcttgca
     5221 gtgggcttac atggcgatag ctagactggg cggttttatg gacagcaagc gaaccggaat
     5281 tgccagctgg ggcgccctct gggaaggttg ggaagccctg caaagtaaac tggatggctt
     5341 tcttgccgcc aaggatctga tggcgcaggg gatcaagatc tgatcaagag acaggggccg
     5401 gctcgctacc ttaggaccgt tatagttagg ggatcaatta ccctgttatc cctaggaagc
     5461 cgatctcggc ttgaacgaat tgttaggtgg cggtacttgg gtcgatatca aagtgcatca
     5521 cttcttcccg tatgcccaac tttgtataga gagccactgc gggatcgtca ccgtaatctg
     5581 cttgcacgta gatcacataa gcaccaagcg cgttggcctc atgcttgagg agattgatga
     5641 gcgcggtggc aatgccctgc ctccggtgct cgccggagac tgcgagatca tagatataga
     5701 tctcactacg cggctgctca aacttgggca gaacgtaagc cgcgagagcg ccaacaaccg
     5761 cttcttggtc gaaggcagca agcgcgatga atgtcttact acggagcaag ttcccgaggt
     5821 aatcggagtc cggctgatgt tgggagtagg tggctacgtc tccgaactca cgaccgaaaa
     5881 gatcaagagc agcccgcatg gatttgactt ggtcagggcc gagcctacat gtgcgaatga
     5941 tgcccatact tgagccacct aactttgttt tagggcgact gccctgctgc gtaacatcgt
     6001 tgctgctgcg taacatcgtt gctgctccat aacatcaaac atcgacccac ggcgtaacgc
     6061 gcttgctgct tggatgcccg aggcatagac tgtacaaaaa aacagtcata acaagccatg
     6121 aaaaccgcca ctgcgccgtt accaccgctg cgttcggtca aggttctgga ccagttgcgt
     6181 gagcgcatac gctacttgca ttacagttta cgaaccgaac aggcttatgt caactgggtt
     6241 cgtgccttca tccgtttcca cggtgtgcgt cggatctatt accctgttat ccctattact
     6301 tgtacagctc gtccatgccg agagtgatcc cggcggcggt cacgaactcc agcaggacca
     6361 tgtgatcgcg cttctcgttg gggtctttgc tcagggcgga ctggtagctc aggtagtggt
     6421 tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga
     6481 gctgcacgct gccgtcctcg atgttgtggc ggatcttgaa gttcaccttg atgccgttct
     6541 tctgcttgtc ggccatgata tagacgttgt ggctgttgta gttgtactcc agcttgtgcc
     6601 ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg
     6661 tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga
     6721 tggtgcgctc ctggacgtag ccttcgggca tggcggactt gaagaagtcg tgctgcttca
     6781 tgtggtcggg gtagcgggcg aagcactgca ggccgtagcc gaaggtggtc acgagggtgg
     6841 gccagggcac gggcagcttg ccggtggtgc agatgaactt cagggtcagc ttgccgtagg
     6901 tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca
     6961 gctcgaccag gatgggcacc accccggtga acagctcctc gcccttgctc accattctcc
     7021 ttaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc ccatctagat
     7081 ctcgagctcg cgaaagcttg gcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     7141 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     7201 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc
     7261 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca
     7321 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     7381 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     7441 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     7501 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     7561 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     7621 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     7681 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     7741 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     7801 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     7861 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     7921 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     7981 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     8041 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     8101 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     8161 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     8221 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     8281 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     8341 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     8401 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     8461 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     8521 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     8581 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     8641 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     8701 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     8761 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     8821 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     8881 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     8941 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     9001 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     9061 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     9121 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     9181 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     9241 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     9301 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     9361 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     9421 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     9481 tgagcgagga agcggaaga
//
