LOCUS       40924_45113        5973 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pUC18T-miniTn7T-LAC-Zeo, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5973)
  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 5973)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 5973)
  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..5973
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     mobile_element  412..610
                     /mobile_element_type="transposon:Tn7R"
                     /label=n7R
     primer_bind     615..631
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      659..753
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      856..942
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     protein_bind    988..1035
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(1148..1519)
                     /label=BleoR
                     /note="antibiotic-binding protein"
     promoter        complement(1538..1585)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     promoter        complement(1764..1792)
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     misc_feature    1833..1880
                     /label=FRT-L
                     /note="FRT-L"
     protein_bind    1833..1880
                     /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(1926..3005)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(3006..3083)
                     /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        3313..3341
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    3349..3365
                     /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)."
     mobile_element  complement(3468..3633)
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     oriT            complement(3792..3900)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      complement(4132..4720)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4894..5751)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5752..5856)
                     /label=AmpR promoter
ORIGIN
        1 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc
       61 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
      121 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
      181 cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat
      241 gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg
      301 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct agaggaccag ccgcgtaacc
      361 tggcaaaatc ggttacggtt gagtaataaa tggatgccct gcgtaagcgg gtgtgggcgg
      421 acaataaagt cttaaactga acaaaataga tctaaactat gacaataaag tcttaaacta
      481 gacagaatag ttgtaaactg aaatcagtcc agttatgctg tgaaaaagca tactggactt
      541 ttgttatggc taaagcaaac tcttcatttt ctgaagtgca aattgcccgt cgtattaaag
      601 aggggcgtgg ggttcgaggt cgacggtatc gataagctag cttaattagc tgagcttgga
      661 ctcctgttga tagatccagt aatgacctca gaactccatc tggatttgtt cagaacgctc
      721 ggttgccgcc gggcgttttt tattggtgag aatccaagct agactgcgat gagtggcagg
      781 gcggggcgta atttttttaa ggcagttatt ggtgccctta aacgcctggg gtaatgactc
      841 tctagcttga ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
      901 atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccgct aggagcttgc
      961 ggcccggacg atcgaattgg ggatcttgaa gtacctattc cgaagttcct attctctaga
     1021 aagtatagga acttcagagc gcttttgaag ctgatgtgct taaaaactta ctcaatggaa
     1081 ttctagatcg aattggccgc ggtctctgat gttaattgca caagataaaa atatatcatc
     1141 atgatcagtc ctgctcctcg gccacgaagt gcacgcagtt gccggccggg tcgcgcaggg
     1201 cgaactcccg cccccacggc tgctcgccga tctcggtcat ggccggcccg gaggcgtccc
     1261 ggaagttcgt ggacacgacc tccgaccact cggcgtacag ctcgtccagg ccgcgcaccc
     1321 acacccaggc cagggtgttg tccggcacca cctggtcctg gaccgcgctg atgaacaggg
     1381 tcacgtcgtc ccggaccaca ccggcgaagt cgtcctccac gaagtcccgg gagaacccga
     1441 gccggtcggt ccagaactcg accgctccgg cgacgtcgcg cgcggtgagc accggaacgg
     1501 cactggtcaa cttggccatg gtttagttcc tcaccttgtc gtattatact atgccgatat
     1561 actatgccga tgattaattg tcaacacgtg ctgatcatga ccaaaatccc ttaacgtgag
     1621 ttacgcgtcg aagccgaata tctagaattt gtacaaaaaa acagtcataa caagccatga
     1681 aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg
     1741 agcgcatacg ctacttgcat tacagtttac gaaccgaaca ggcttatgtc aattcgatcc
     1801 attgctgttg acaaagggaa tcaggggatc ttgaagttcc tattccgaag ttcctattct
     1861 ctagaaagta taggaacttc agagcgcttt tgaagctaat tcgatcatgc attgcgttgc
     1921 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     1981 aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt
     2041 gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg
     2101 tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata
     2161 taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatatccgc accaacgcgc
     2221 agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc
     2281 atcgcagtag gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg
     2341 gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta
     2401 tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg
     2461 atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg
     2521 gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca
     2581 ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc
     2641 agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg
     2701 cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc
     2761 gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc
     2821 aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc
     2881 atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg
     2941 cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt
     3001 ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag
     3061 gttttgcacc attcgatggt gtcaacgtaa atgccgcttc gccttcgcgc gcgaattgca
     3121 agctgatccg ggcttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc
     3181 ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc
     3241 gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc
     3301 tgaaatgagc tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat
     3361 aacaatttca cacaggaaac agaattcgag ctcactagtg gatcccccgg gctgcaggaa
     3421 ttcctcgaga agcttgggcc cggtacctcg cgaaggcctt gcaggccaac cagataagtg
     3481 aaatctagtt ccaaactatt ttgtcatttt taattttcgt attagcttac gacgctacac
     3541 ccagttccca tctattttgt cactcttccc taaataatcc ttaaaaactc catttccacc
     3601 cctcccagtt cccaactatt ttgtccgccc acagcggggc atttttcttc ctgttatgtt
     3661 tgggcgctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     3721 tcttccgctt cctcgctcac tgacccgctg cgctcggtcg ttcggctgcg gcgagcggta
     3781 tcagagctta tcggccagcc tcgcagagca ggattcccgt tgagcaccgc caggtgcgaa
     3841 taagggacag tgaagaagga acacccgctc gcgggtgggc ctacttcacc tatcctgccc
     3901 ggctgacgcc gttggataca ccaaggaaag tctacacgaa ccctttggca aaatcctgta
     3961 tatcgtgcga aaaaggatgg atataccgaa aaaatcgcta taatgacccc gaagcagggt
     4021 tatgcagcgg aaagtatacc ttaaggaatc ccctgataac gcaggaaaga acatgtgagc
     4081 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4141 gctccgcccc cctgacgagc atcacaaaaa tcgacgctcg agtcagaggt ggcgaaaccc
     4201 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4261 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4321 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4381 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4441 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4501 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4561 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4621 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4681 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4741 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4801 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     4861 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     4921 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4981 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     5041 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5101 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5161 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5221 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5281 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     5341 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     5401 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     5461 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     5521 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     5581 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5641 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5701 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     5761 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     5821 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     5881 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     5941 gccctttcgt ctcgcgcgtt tcggtgatga cgg
//
