LOCUS       40924_45058        4871 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pUC18T-mini-Tn7T-aadA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4871)
  AUTHORS   Lehman SS, Mladinich K, Boonyakanog A, Mima T, Karkhoff-Schweizer 
            RR, Schweizer HP.
  TITLE     Versatile nourseothricin and streptomycin/spectinomycin resistance 
            gene cassettes and their use in chromosome integration vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4871)
  AUTHORS   Lehman SS, Mladinich K, Boonyakanog A, Mima T, Karkhoff-Schweizer 
            RR, Schweizer HP.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-MAR-2016) Molecular Genetics and Microbiology, 
            University of Florida, 2055 Mowry Road, Gainesville, FL 32610, USA
REFERENCE   3  (bases 1 to 4871)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4871)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (17-MAR-2016) Molecular Genetics and Microbiology, University of 
            Florida, 2055 Mowry Road, Gainesville, FL 32610, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4871
                     /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"
     misc_feature    955..988
                     /note="loxP; Cre recombinase site"
     CDS             complement(1162..1950)
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
     regulatory      2052..2101
                     /label=aadA gene promoter
                     /note="aadA gene promoter"
                     /regulatory_class="promoter"
     protein_bind    complement(2254..2287)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2302..2376
                     /note="MCS; multiple cloning site"
     mobile_element  complement(2388..2553)
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     oriT            complement(2712..2820)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      complement(3052..3640)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3814..4671)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4672..4776)
                     /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 atgggtaccc gcgtctactt
      961 cgtatagcat acattatacg aagttatact aaaacaattc atccagtaaa atataatatt
     1021 ttattttctc ccaatcaggc ttgatcccga attcggcttc ctgatagttt ggctgtgagc
     1081 aattatgtgc ttagtgcatc taacgcttga gttaagccgc gccgcgaagc ggcgtcggct
     1141 tgaacgaatt gttagacatt atttgccgac taccttggtg atctcgcctt tcacgtagtg
     1201 gacaaattct tccaactgat ctgcgcgcga ggccaagcga tcttcttctt gtccaagata
     1261 agcctgtcta gcttcaagta tgacgggctg atactgggcc ggcaggcgct ccattgccca
     1321 gtcggcagcg acatccttcg gcgcgatttt gccggttact gcgctgtacc aaatgcggga
     1381 caacgtaagc actacatttc gctcatcgcc agcccagtcg ggcggcgagt tccatagcgt
     1441 taaggtttca tttagcgcct caaatagatc ctgttcagga accggatcaa agagttcctc
     1501 cgccgctgga cctaccaagg caacgctatg ttctcttgct tttgtcagca agatagccag
     1561 atcaatgtcg atcgtggctg gctcgaagat acctgcaaga atgtcattgc gctgccattc
     1621 tccaaattgc agttcgcgct tagctggata acgccacgga atgatgtcgt cgtgcacaac
     1681 aatggtgact tctacagcgc ggagaatctc gctctctcca ggggaagccg aagtttccaa
     1741 aaggtcgttg atcaaagctc gccgcgttgt ttcatcaagc cttacggtca ccgtaaccag
     1801 caaatcaata tcactgtgtg gcttcaggcc gccatccact gcggagccgt acaaatgtac
     1861 ggccagcaac gtcggttcga gatggcgctc gatgacgcca actacctctg atagttgagt
     1921 cgatacttcg gcgatcaccg cttccctcat gatgtttaac tttgttttag ggcgactgcc
     1981 ctgctgcgta acatcgttgc tgctccataa catcaaacat cgacccacgg cgtaacgcgc
     2041 ttgctgcttg gatgcccgag gcatagactg taccccaaaa aaacagtcat aacaagccat
     2101 gaaaaccgcc actgcgccgt taccaccgct gcgttcggtc aaggttctgg accagttgcg
     2161 tgagcgcata cgctacttgc attacagctt acgaaccgaa caggcttatg tccactgggt
     2221 tcgaagccga attccgacgg agccgatttt gaaataactt cgtatagcat acattatacg
     2281 aagttatacg cggtacccat catgcatgag ctcactagtg gatcccccgg gctgcaggaa
     2341 ttcctcgaga agcttgggcc cggtacctcg cgaaggcctt gcaggccaac cagataagtg
     2401 aaatctagtt ccaaactatt ttgtcatttt taattttcgt attagcttac gacgctacac
     2461 ccagttccca tctattttgt cactcttccc taaataatcc ttaaaaactc catttccacc
     2521 cctcccagtt cccaactatt ttgtccgccc acagcggggc atttttcttc ctgttatgtt
     2581 tgggcgctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     2641 tcttccgctt cctcgctcac tgacccgctg cgctcggtcg ttcggctgcg gcgagcggta
     2701 tcagagctta tcggccagcc tcgcagagca ggattcccgt tgagcaccgc caggtgcgaa
     2761 taagggacag tgaagaagga acacccgctc gcgggtgggc ctacttcacc tatcctgccc
     2821 ggctgacgcc gttggataca ccaaggaaag tctacacgaa ccctttggca aaatcctgta
     2881 tatcgtgcga aaaaggatgg atataccgaa aaaatcgcta taatgacccc gaagcagggt
     2941 tatgcagcgg aaagtatacc ttaaggaatc ccctgataac gcaggaaaga acatgtgagc
     3001 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3061 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3121 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3181 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     3241 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     3301 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3361 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     3421 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3481 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     3541 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     3601 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     3661 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     3721 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     3781 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     3841 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     3901 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     3961 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     4021 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     4081 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     4141 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     4201 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     4261 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     4321 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     4381 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     4441 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     4501 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     4561 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     4621 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     4681 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     4741 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     4801 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     4861 gccctttcgt c
//
