LOCUS       62056_21955        5670 bp DNA     circular SYN 02-APR-2019
DEFINITION  Cloning vector pUC18T-mini-Tn7T-Apr-sfGFP, complete sequence.
ACCESSION   MH976505
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5670)
  AUTHORS   Ducas-Mowchun K, De Silva PM, Crisostomo L, Fernando DM, Chao TC, 
            Pelka P, Schweizer HP, Kumar A.
  TITLE     Next Generation of Tn7-based single copy insertion elements for use 
            in multi- and pandrug resistant strains of Acinetobacter baumannii
  JOURNAL   Appl. Environ. Microbiol. (2019) In press
  PUBMED    30902859
REFERENCE   2  (bases 1 to 5670)
  AUTHORS   Ducas-Mowchun K, De Silva PM, Kumar A.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-SEP-2018) Department of Microbiology, University of 
            Manitoba, 45, Chancellor Circle, Winnipeg, Manitoba R3T2N2, Canada
REFERENCE   3  (bases 1 to 5670)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Environ. Microbiol. (2019) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (24-SEP-2018) Department of Microbiology, University of Manitoba, 
            45, Chancellor Circle, Winnipeg, Manitoba R3T2N2, Canada"
FEATURES             Location/Qualifiers
     source          1..5670
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     mobile_element  382..580
                     /mobile_element_type="transposon:Tn7R"
                     /label=n7R
                     /note="right end of Tn7"
     primer_bind     585..601
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      629..723
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      826..912
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     protein_bind    958..1005
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(1123..1923)
                     /label=ApmR
                     /note="aminoglycoside 3-N-acetyltransferase type IV"
     promoter        complement(2164..2192)
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     misc_feature    2233..2280
                     /note="FRT; Flp recombinase target site"
     misc_feature    2307..2355
                     /note="MCS; multiple cloning site"
     CDS             complement(2364..3077)
                     /label=superfolder GFP
                     /note="GFP variant that folds robustly even when fused to
                     poorly folded proteins (Nager et al., 2011)"
     protein_bind    complement(3108..3124)
                     /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(3132..3160)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     misc_feature    3166..3178
                     /note="MCS; multiple cloning site"
     mobile_element  complement(3187..3352)
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     oriT            complement(3511..3619)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      complement(3851..4439)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4613..5470)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5471..5575)
                     /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 ggttcgaggt cgacggtatc
      601 gataagctag cttaattagc tgagcttgga ctcctgttga tagatccagt aatgacctca
      661 gaactccatc tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggtgag
      721 aatccaagct agactgcgat gagtggcagg gcggggcgta atttttttaa ggcagttatt
      781 ggtgccctta aacgcctggg gtaatgactc tctagcttga ggcatcaaat aaaacgaaag
      841 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg
      901 agtaggacaa atccgccgct aggagcttgc ggcccggacg atcgaattgg ggatcttgaa
      961 gttcctattc cgaagttcct attctctaga aagtatagga acttcagagc gcttttgaag
     1021 ctgatgtgct taaaaactta ctcaatggaa ttctagatcg aattggccgc ggtagataat
     1081 tcggcttctc caacgtcatc tcgttctccg ctcatgagct cagccaatcg actggcgagc
     1141 ggcatcgcat tcttcgcatc ccgcctctgg cggatgcagg aagatcaacg gatctcggcc
     1201 cagttgaccc agggctgtcg ccacaatgtc gcgggagcgg atcaaccgag caaaggcatg
     1261 accgactgga ccttccttct gaaggctctt ctccttgagc cacctgtccg ccaaggcaaa
     1321 gcgctcacag cagtggtcat tctcgagata atcgacgcgt accaacttgc catcctgaag
     1381 aatggtgcag tgtctcggca ccccataggg aacctttgcc atcaactcgg caagatgcag
     1441 cgtcgtgttg gcatcgtgtc ccacgccgag gagaagtacc tgcccatcga gttcatggac
     1501 acgggcgacc gggcttgcag gcgagtgagg tggcaggggc aatggatcag agatgatctg
     1561 ctctgcctgt ggccccgctg ccgcaaaggc aaatggatgg gcgctgcgct ttacatttgg
     1621 caggcgccag aatgtgtcag agataactcc aaggtccggt gtaacgggcg acgtggcagg
     1681 atcgaacggc tcgtcgtcca gacctgacca cgagggcatg acgagcgtcc ctcccggacc
     1741 cagcgcagca cgcagggcct cgatcagtcc aagtggccca tcttcgaggg gccggacgct
     1801 acggaaggag ctgtggacca gcagcacacc gccgggggta accccaaggt tgagaagctg
     1861 accgatgagc tcggcttttc gccattcgta ttgcacgaca ttgcactcca ccgctgatga
     1921 catcagtcga tcatagcacg atcaacggca ctgttgcaaa tagtcggtgg tgataaactt
     1981 atcatcccct tttgctgatg gagctgcaca tgaaaagccg aattatccat tgctgttgac
     2041 aaagggaatt gtacaaaaaa acagtcataa caagccatga aaaccgccac tgcgccgtta
     2101 ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg agcgcatacg ctacttgcat
     2161 tacagtttac gaaccgaaca ggcttatgtc aattcgatcc attgctgttg acaaagggaa
     2221 tcaggggatc ttgaagttcc tattccgaag ttcctattct ctagaaagta taggaacttc
     2281 agagcgcttt tgaagctaat tcgatcatgc atgagctcac tagtggatcc cccgggctgc
     2341 aggaattcct cgagaagctt ttatttgtag agctcatcca tgccgtgcgt gatacctgct
     2401 gcagtaacga actccagcag caccatgtgg tcgcgctttt cgttcgggtc tttggacaga
     2461 acagactggg tggacaggta gtggttatcc ggcagcagaa ccggaccatc accgatcgga
     2521 gtgttctgct ggtagtggtc cgccagctgt acgctaccgt cttcaacgtt atggcgaatt
     2581 ttgaagttag ctttgatacc gttcttctgt ttgtctgcgg tgatgtaaac gttatgggag
     2641 ttgaagttat attccagttt gtggcccagg atgttgccgt cctctttgaa atcaatgcct
     2701 ttcagttcaa tacggttcac cagagtatca ccttcaaatt taacctctgc acgggttttg
     2761 taggtgccat cgtctttgaa agaaatggtg cgctcctgta cataaccttc cggcattgca
     2821 gatttgaaga aatcatgctg cttcatgtga tccgggtaac gagaaaaaca ctgaacacca
     2881 taggtcaggg tagtcaccag agtcggccat ggaaccggca gtttaccggt agtgcagatg
     2941 aatttcaggg tcagtttacc gttggttgca tcaccttcac cttcaccacg aacagagaat
     3001 ttgtggccgt taacatcacc atccagttca accaggatcg gaacaacacc ggtgaacagt
     3061 tcttcacctt tagacatgcg gccgcgaatt ctgtttcctg tgtgaaattg ttatccgctc
     3121 acaattccac acattatacg agccgatgat taattgtcaa ggtacctcgc gaaggccttg
     3181 caggccaacc agataagtga aatctagttc caaactattt tgtcattttt aattttcgta
     3241 ttagcttacg acgctacacc cagttcccat ctattttgtc actcttccct aaataatcct
     3301 taaaaactcc atttccaccc ctcccagttc ccaactattt tgtccgccca cagcggggca
     3361 tttttcttcc tgttatgttt gggcgctgca ttaatgaatc ggccaacgcg cggggagagg
     3421 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gacccgctgc gctcggtcgt
     3481 tcggctgcgg cgagcggtat cagagcttat cggccagcct cgcagagcag gattcccgtt
     3541 gagcaccgcc aggtgcgaat aagggacagt gaagaaggaa cacccgctcg cgggtgggcc
     3601 tacttcacct atcctgcccg gctgacgccg ttggatacac caaggaaagt ctacacgaac
     3661 cctttggcaa aatcctgtat atcgtgcgaa aaaggatgga tataccgaaa aaatcgctat
     3721 aatgaccccg aagcagggtt atgcagcgga aagtatacct taaggaatcc cctgataacg
     3781 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     3841 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcga
     3901 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     3961 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4021 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4081 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4141 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4201 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4261 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     4321 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     4381 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     4441 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     4501 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     4561 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     4621 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     4681 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     4741 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     4801 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     4861 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     4921 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     4981 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     5041 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     5101 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     5161 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     5221 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     5281 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     5341 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     5401 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     5461 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     5521 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     5581 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     5641 aaaataggcg tatcacgagg ccctttcgtc
//
