LOCUS       40924_45108        6403 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pUC18T-miniTn7T-LAC-Apr, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6403)
  AUTHORS   Ducas-Mowchun K, DeSilva PM, Kumar A.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-MAY-2018) Department of Microbiology, University of 
            Manitoba, 45, Chancellor Circle, Buller Building (Room 412), 
            Winnipeg, MB R3T2N2, Canada
REFERENCE   2  (bases 1 to 6403)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 6403)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (02-MAY-2018) Department of Microbiology, University of Manitoba, 
            45, Chancellor Circle, Buller Building (Room 412), Winnipeg, MB 
            R3T2N2, Canada"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: Geneious v. R11
            Sequencing Technology :: Illumina
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6403
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     mobile_element  383..581
                     /mobile_element_type="transposon:Tn7R"
                     /label=n7R
     primer_bind     586..602
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      630..724
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      827..913
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     protein_bind    959..1006
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(1124..1924)
                     /label=ApmR
                     /note="aminoglycoside 3-N-acetyltransferase type IV"
     promoter        complement(2165..2193)
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     misc_feature    2234..2281
                     /label=FRT
                     /note="FRT"
     protein_bind    2234..2281
                     /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(2327..3406)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(3407..3484)
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     promoter        3714..3742
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    3750..3766
                     /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)."
     misc_feature    3780..3860
                     /label=MCS
                     /note="MCS"
     mobile_element  complement(3869..4034)
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     oriT            complement(4193..4301)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      complement(4533..5121)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5295..6152)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6153..6257)
                     /label=AmpR promoter
ORIGIN
        1 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
       61 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
      121 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
      181 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
      241 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
      301 ttacgccagc tagaggacca gccgcgtaac ctggcaaaat cggttacggt tgagtaataa
      361 atggatgccc tgcgtaagcg ggtgtgggcg gacaataaag tcttaaactg aacaaaatag
      421 atctaaacta tgacaataaa gtcttaaact agacagaata gttgtaaact gaaatcagtc
      481 cagttatgct gtgaaaaagc atactggact tttgttatgg ctaaagcaaa ctcttcattt
      541 tctgaagtgc aaattgcccg tcgtattaaa gaggggcgtg gggttcgagg tcgacggtat
      601 cgataagcta gcttaattag ctgagcttgg actcctgttg atagatccag taatgacctc
      661 agaactccat ctggatttgt tcagaacgct cggttgccgc cgggcgtttt ttattggtga
      721 gaatccaagc tagactgcga tgagtggcag ggcggggcgt aattttttta aggcagttat
      781 tggtgccctt aaacgcctgg ggtaatgact ctctagcttg aggcatcaaa taaaacgaaa
      841 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct
      901 gagtaggaca aatccgccgc taggagcttg cggcccggac gatcgaattg gggatcttga
      961 agtacctatt ccgaagttcc tattctctag aaagtatagg aacttcagag cgcttttgaa
     1021 gctgatgtgc ttaaaaactt actcaatgga attctagatc gaattggccg cggtagataa
     1081 ttcggcttct ccaacgtcat ctcgttctcc gctcatgagc tcagccaatc gactggcgag
     1141 cggcatcgca ttcttcgcat cccgcctctg gcggatgcag gaagatcaac ggatctcggc
     1201 ccagttgacc cagggctgtc gccacaatgt cgcgggagcg gatcaaccga gcaaaggcat
     1261 gaccgactgg accttccttc tgaaggctct tctccttgag ccacctgtcc gccaaggcaa
     1321 agcgctcaca gcagtggtca ttctcgagat aatcgacgcg taccaacttg ccatcctgaa
     1381 gaatggtgca gtgtctcggc accccatagg gaacctttgc catcaactcg gcaagatgca
     1441 gcgtcgtgtt ggcatcgtgt cccacgccga ggagaagtac ctgcccatcg agttcatgga
     1501 cacgggcgac cgggcttgca ggcgagtgag gtggcagggg caatggatca gagatgatct
     1561 gctctgcctg tggccccgct gccgcaaagg caaatggatg ggcgctgcgc tttacatttg
     1621 gcaggcgcca gaatgtgtca gagataactc caaggtccgg tgtaacgggc gacgtggcag
     1681 gatcgaacgg ctcgtcgtcc agacctgacc acgagggcat gacgagcgtc cctcccggac
     1741 ccagcgcagc acgcagggcc tcgatcagtc caagtggccc atcttcgagg ggccggacgc
     1801 tacggaagga gctgtggacc agcagcacac cgccgggggt aaccccaagg ttgagaagct
     1861 gaccgatgag ctcggctttt cgccattcgt attgcacgac attgcactcc accgctgatg
     1921 acatcagtcg atcatagcac gatcaacggc actgttgcaa atagtcggtg gtgataaact
     1981 tatcatcccc ttttgctgat ggagctgcac atgaaaagcc gaattatcca ttgctgttga
     2041 caaagggaat tgtacaaaaa aacagtcata acaagccatg aaaaccgcca ctgcgccgtt
     2101 accaccgctg cgttcggtca aggttctgga ccagttgcgt gagcgcatac gctacttgca
     2161 ttacagttta cgaaccgaac aggcttatgt caattcgatc cattgctgtt gacaaaggga
     2221 atcaggggat cttgaagttc ctattccgaa gttcctattc tctagaaagt ataggaactt
     2281 cagagcgctt ttgaagctaa ttcgatcatg cattgcgttg cgctcactgc ccgctttcca
     2341 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     2401 tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat
     2461 tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca
     2521 gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg
     2581 tatcgtcgta tcccactacc gagatatccg caccaacgcg cagcccggac tcggtaatgg
     2641 cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagta ggaacgatgc
     2701 cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc
     2761 gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca
     2821 gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg
     2881 cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga
     2941 tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca
     3001 cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg
     3061 cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca
     3121 ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg
     3181 gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca
     3241 gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt
     3301 cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag
     3361 agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc accaccctga
     3421 attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcac cattcgatgg
     3481 tgtcaacgta aatgccgctt cgccttcgcg cgcgaattgc aagctgatcc gggcttatcg
     3541 actgcacggt gcaccaatgc ttctggcgtc aggcagccat cggaagctgt ggtatggctg
     3601 tgcaggtcgt aaatcactgc ataattcgtg tcgctcaagg cgcactcccg ttctggataa
     3661 tgttttttgc gccgacatca taacggttct ggcaaatatt ctgaaatgag ctgttgacaa
     3721 ttaatcatcg gctcgtataa tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     3781 cagaattcga gctcactagt ggatcccccg ggctgcagga attcctcgag aagcttgggc
     3841 ccggtacctc gcgaaggcct tgcaggccaa ccagataagt gaaatctagt tccaaactat
     3901 tttgtcattt ttaattttcg tattagctta cgacgctaca cccagttccc atctattttg
     3961 tcactcttcc ctaaataatc cttaaaaact ccatttccac ccctcccagt tcccaactat
     4021 tttgtccgcc cacagcgggg catttttctt cctgttatgt ttgggcgctg cattaatgaa
     4081 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     4141 ctgacccgct gcgctcggtc gttcggctgc ggcgagcggt atcagagctt atcggccagc
     4201 ctcgcagagc aggattcccg ttgagcaccg ccaggtgcga ataagggaca gtgaagaagg
     4261 aacacccgct cgcgggtggg cctacttcac ctatcctgcc cggctgacgc cgttggatac
     4321 accaaggaaa gtctacacga accctttggc aaaatcctgt atatcgtgcg aaaaaggatg
     4381 gatataccga aaaaatcgct ataatgaccc cgaagcaggg ttatgcagcg gaaagtatac
     4441 cttaaggaat cccctgataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4501 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4561 catcacaaaa atcgacgctc gagtcagagg tggcgaaacc cgacaggact ataaagatac
     4621 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4681 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4741 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4801 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4861 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4921 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     4981 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5041 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5101 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5161 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5221 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5281 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5341 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5401 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     5461 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     5521 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     5581 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     5641 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5701 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5761 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5821 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     5881 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     5941 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6001 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6061 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6121 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6181 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6241 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     6301 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     6361 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cgg
//
