LOCUS       40924_44973        8347 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pUC18-mini-Tn7-LACM15, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8347)
  AUTHORS   Choi KH, Gaynor JB, White KG, Lopez C, Bosio CM, Karkhoff-Schweizer 
            RR, Schweizer HP.
  TITLE     A Tn7-based broad-range bacterial cloning and expression system
  JOURNAL   Nat. Methods 2 (6), 443-448 (2005)
  PUBMED    15908923
REFERENCE   2  (bases 1 to 8347)
  AUTHORS   Choi K-H., Gaynor JB, Karkhoff-Schweizer RR, Schweizer HP.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-MAY-2004) Microbiology, Immunology and Pathology, 
            Colorado State University, Fort Collins, CO 80523, USA
REFERENCE   3  (bases 1 to 8347)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8347)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat. 
            Methods"; date: "2005"; volume: "2"; issue: "6"; pages: "443-448"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (06-MAY-2004) Microbiology, Immunology and Pathology, Colorado State
            University, Fort Collins, CO 80523, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8347
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   382..580
                     /note="Tn7R; right end of Tn7"
     protein_bind    600..647
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(780..1310)
                     /label=GmR
                     /note="gentamycin acetyltransferase"
     promoter        complement(1499..1527)
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     misc_feature    1568..1615
                     /label=FRT, Flp recombinase target site
                     /note="FRT, Flp recombinase target site"
     protein_bind    1568..1615
                     /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(1765..4710)
                     /label=lacZ
                     /note="beta-galactosidase"
     primer_bind     complement(4739..4755)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4763..4779)
                     /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)."
     promoter        complement(4787..4817)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4832..4853)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(4869..5948)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(5949..6026)
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     mobile_element  complement(6093..6258)
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      complement(6528..7116)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7290..8147)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(8148..8252)
                     /label=AmpR promoter
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct agaggaccag ccgcgtaacc tggcaaaatc ggttacggtt gagtaataaa
      361 tggatgccct gcgtaagcgg gtgtgggcgg acaataaagt cttaaactga acaaaataga
      421 tctaaactat gacaataaag tcttaaacta gacagaatag ttgtaaactg aaatcagtcc
      481 agttatgctg tgaaaaagca tactggactt ttgttatggc taaagcaaac tcttcatttt
      541 ctgaagtgca aattgcccgt cgtattaaag aggggcgtgg ggtccgaatt ggggatcttg
      601 aagtacctat tccgaagttc ctattctcta gaaagtatag gaacttcaga gcgcttttga
      661 agctgatgtg cttaaaaact tactcaatgg aattctagat cgaattggcc gcggcgttgt
      721 gacaatttac cgaacaactc cgcggccggg aagccgatct cggcttgaac gaattgttag
      781 gtggcggtac ttgggtcgat atcaaagtgc atcacttctt cccgtatgcc caactttgta
      841 tagagagcca ctgcgggatc gtcaccgtaa tctgcttgca cgtagatcac ataagcacca
      901 agcgcgttgg cctcatgctt gaggagattg atgagcgcgg tggcaatgcc ctgcctccgg
      961 tgctcgccgg agactgcgag atcatagata tagatctcac tacgcggctg ctcaaacttg
     1021 ggcagaacgt aagccgcgag agcgccaaca accgcttctt ggtcgaaggc agcaagcgcg
     1081 atgaatgtct tactacggag caagttcccg aggtaatcgg agtccggctg atgttgggag
     1141 taggtggcta cgtctccgaa ctcacgaccg aaaagatcaa gagcagcccg catggatttg
     1201 acttggtcag ggccgagcct acatgtgcga atgatgccca tacttgagcc acctaacttt
     1261 gttttagggc gactgccctg ctgcgtaaca tcgttgctgc tgcgtaacat cgttgctgct
     1321 ccataacatc aaacatcgac ccacggcgta acgcgcttgc tgcttggatg cccgaggcat
     1381 agactgtaca aaaaaacagt cataacaagc catgaaaacc gccactgcgc cgttaccacc
     1441 gctgcgttcg gtcaaggttc tggaccagtt gcgtgagcgc atacgctact tgcattacag
     1501 tttacgaacc gaacaggctt atgtcaattc gatccattgc tgttgacaaa gggaatcagg
     1561 ggatcttgaa gttcctattc cgaagttcct attctctaga aagtatagga acttcagagc
     1621 gcttttgaag ctaattcgga cgatatcatg catgagctca ctagtggatc ccccgggctg
     1681 caggaattcg agctcgccca aatagtacat aatggatttc cttacgcgaa atacgggcag
     1741 acatggcctg cccggttatt attatttttg acaccagacc aactggtaat ggtagcgacc
     1801 ggcgctcagc tggaattccg ccgatactga cgggctccag gagtcgtcgc caccaatccc
     1861 catatggaaa ccgtcgatat tcagccatgt gccttcttcc gcgtgcagca gatggcgatg
     1921 gctggtttcc atcagttgct gttgactgta gcggctgatg ttgaactgga agtcgccgcg
     1981 ccactggtgt gggccataat tcaattcgcg cgtcccgcag cgcagaccgt tttcgctcgg
     2041 gaagacgtac ggggtataca tgtctgacaa tggcagatcc cagcggtcaa aacaggcggc
     2101 agtaaggcgg tcgggatagt tttcttgcgg ccctaatccg agccagttta cccgctctgc
     2161 tacctgcgcc agctggcagt tcaggccaat ccgcgccgga tgcggtgtat cgctcgccac
     2221 ttcaacatca acggtaatcg ccatttgacc actaccatca atccggtagg ttttccggct
     2281 gataaataag gttttcccct gatgctgcca cgcgtgagcg gtcgtaatca gcaccgcatc
     2341 agcaagtgta tctgccgtgc actgcaacaa cgctgcttcg gcctggtaat ggcccgccgc
     2401 cttccagcgt tcgacccagg cgttagggtc aatgcgggtc gcttcactta cgccaatgtc
     2461 gttatccagc ggtgcacggg tgaactgatc gcgcagcggc gtcagcagtt gttttttatc
     2521 gccaatccac atctgtgaaa gaaagcctga ctggcggtta aattgccaac gcttattacc
     2581 cagctcgatg caaaaatcca tttcgctggt ggtcagatgc gggatggcgt gggacgcggc
     2641 ggggagcgtc acactgaggt tttccgccag acgccactgc tgccaggcgc tgatgtgccc
     2701 ggcttctgac catgcggtcg cgttcggttg cactacgcgt actgtgagcc agagttgccc
     2761 ggcgctctcc ggctgcggta gttcaggcag ttcaatcaac tgtttacctt gtggagcgac
     2821 atccagaggc acttcaccgc ttgccagcgg cttaccatcc agcgccacca tccagtgcag
     2881 gagctcgtta tcgctatgac ggaacaggta ttcgctggtc acttcgatgg tttgcccgga
     2941 taaacggaac tggaaaaact gctgctggtg ttttgcttcc gtcagcgctg gatgcggcgt
     3001 gcggtcggca aagaccagac cgttcataca gaactggcga tcgttcggcg tatcgccaaa
     3061 atcaccgccg taagccgacc acgggttgcc gttttcatca tatttaatca gcgactgatc
     3121 cacccagtcc cagacgaagc cgccctgtaa acggggatac tgacgaaacg cctgccagta
     3181 tttagcgaaa ccgccaagac tgttacccat cgcgtgggcg tattcgcaaa ggatcagcgg
     3241 gcgcgtctct ccaggtagcg aaagccattt tttgatggac catttcggca cagccgggaa
     3301 gggctggtct tcatccacgc gcgcgtacat cgggcaaata atatcggtgg ccgtggtgtc
     3361 ggctccgccg ccttcatact gcaccgggcg ggaaggatcg acagatttga tccagcgata
     3421 cagcgcgtcg tgattagcgc cgtggcctga ttcattcccc agcgaccaga tgatcacact
     3481 cgggtgatta cgatcgcgct gcaccattcg cgttacgcgt tcgctcatcg ccggtagcca
     3541 gcgcggatca tcggtcagac gattcattgg caccatgccg tgggtttcaa tattggcttc
     3601 atccaccaca tacaggccgt agcggtcgca cagcgtgtac cacagcggat ggttcggata
     3661 atgcgaacag cgcacggcgt taaagttgtt ctgcttcatc agcaggatat cctgcaccat
     3721 cgtctgctca tccatgacct gaccatgcag aggatgatgc tcgtgacggt taacgcctcg
     3781 aatcagcaac ggcttgccgt tcagcagcag cagaccattt tcaatccgca cctcgcggaa
     3841 accgacatcg caggcttctg cttcaatcag cgtgccgtcg gcggtgtgca gttcaaccac
     3901 cgcacgatag agattcggga tttcggcgct ccacagtttc gggttttcga cgttcagacg
     3961 tagtgtgacg cgatcggcat aaccaccacg ctcatcgata atttcaccgc cgaaaggcgc
     4021 ggtgccgctg gcgacctgcg tttcaccctg ccataaagaa actgttaccc gtaggtagtc
     4081 acgcaactcg ccgcacatct gaacttcagc ctccagtaca gcgcggctga aatcatcatt
     4141 aaagcgagtg gcaacatgga aatcgctgat ttgtgtagtc ggtttatgca gcaacgagac
     4201 gtcacggaaa atgccgctca tccgccacat atcctgatct tccagataac tgccgtcact
     4261 ccaacgcagc accatcaccg cgaggcggtt ttctccggcg cgtaaaaatg cgctcaggtc
     4321 aaattcagac ggcaaacgac tgtcctggcc gtaaccgacc cagcgcccgt tgcaccacag
     4381 atgaaacgcc gagttaacgc catcaaaaat aattcgcgtc tggccttcct gtagccagct
     4441 ttcatcaaca ttaaatgtga gcgagtaaca acccgtcgga ttctccgtgg gaacaaacgg
     4501 cggattgacc gtaatgggat aggttacgtt ggtgtagatg ggcgcatcgt aaccgtgcat
     4561 ctgccagttt gaggggacga cgacagtatc ggcctcagga agatcgcact ccagccagct
     4621 ttccggcacc gcttctggtg ccggaaacca ggcaaagcgc cattcgccat tcaggctgcg
     4681 caactgttgg gaagggcgat cggtgcgggc cacgacggcc agtgaatccg taatcatggt
     4741 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     4801 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     4861 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     4921 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc
     4981 agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag
     5041 cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt tgacggcggg
     5101 atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg
     5161 cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc
     5221 agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac
     5281 atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat
     5341 ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc
     5401 gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca
     5461 tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga
     5521 acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg
     5581 atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg
     5641 ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta
     5701 atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc
     5761 agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc
     5821 gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc
     5881 acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact
     5941 ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga
     6001 aaggttttgc accattcgat ggtgtcagga tcctctagag tcgacctgca gcccaagctt
     6061 gggcccggta cctcgcgaag gccttgcagg ccaaccagat aagtgaaatc tagttccaaa
     6121 ctattttgtc atttttaatt ttcgtattag cttacgacgc tacacccagt tcccatctat
     6181 tttgtcactc ttccctaaat aatccttaaa aactccattt ccacccctcc cagttcccaa
     6241 ctattttgtc cgcccacagc ggggcatttt tcttcctgtt atgtttgggc gctgcattaa
     6301 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     6361 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     6421 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     6481 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     6541 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     6601 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     6661 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     6721 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6781 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6841 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6901 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6961 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     7021 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     7081 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     7141 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     7201 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     7261 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     7321 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     7381 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     7441 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     7501 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     7561 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     7621 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     7681 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     7741 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     7801 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     7861 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     7921 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     7981 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     8041 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     8101 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     8161 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     8221 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     8281 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     8341 tttcgtc
//
