LOCUS       40924_45083        4838 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pUC18T-mini-Tn7T-nat, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4838)
  AUTHORS   Lehman SS, Mladinich KM, 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   J. Microbiol. Methods 129, 8-13 (2016)
  PUBMED    27457407
REFERENCE   2  (bases 1 to 4838)
  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 4838)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4838)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J. 
            Microbiol. Methods 129, 8-13 (2016)"
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..4838
                     /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    965..987
                     /note="loxP; Cre recombinase site"
     promoter        1117..1460
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1467..2030
                     /label=NrsR
                     /note="nourseothricin acetyltransferase"
     protein_bind    complement(2210..2243)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2269..2343
                     /note="MCS; multiple cloning site"
     mobile_element  complement(2355..2520)
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     oriT            complement(2679..2787)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      complement(3019..3607)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3781..4638)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4639..4743)
                     /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 atctagagga tccccgggta
      961 ccgtctactt cgtatagcat acattatacg aagttatact aaaacaattc atccagtaaa
     1021 atataatatt ttattttctc ccaatcaggc ttgatcccga attcggcttc cagatctgtt
     1081 tagcttgcct tgtccccgcc gggtcacccg gccagcgaca tggaggccca gaataccctc
     1141 cttgacagtc ttgacgtgcg cagctcaggg gcatgatgtg actgtcgccc gtacatttag
     1201 cccatacatc cccatgtata atcatttgca tccatacatt ttgatggccg cacggcgcga
     1261 agcaaaaatt acggctcctc gctgcagacc tgcgagcagg gaaacgctcc cctcacagac
     1321 gcgttgaatt gtccccacgc cgcgcccctg tagagaaata taaaaggtta ggatttgcca
     1381 ctgaggttct tctttcatat acttcctttt aaaatcttgc taggatacag ttctcacatc
     1441 acatccgaac ataaacaacc atgggtacca ctcttgacga cacggcttac cggtaccgca
     1501 ccagtgtccc gggggacgcc gaggccatcg aggcactgga tgggtccttc accaccgaca
     1561 ccgtcttccg cgtcaccgcc accggggacg gcttcaccct gcgggaggtg ccggtggacc
     1621 cgcccctgac caaggtgttc cccgacgacg aatcggacga cgaatcggac gacggggagg
     1681 acggcgaccc ggactcccgg acgttcgtcg cgtacgggga cgacggcgac ctggcgggct
     1741 tcgtggtcgt ctcgtactcc ggctggaacc gccggctgac cgtcgaggac atcgaggtcg
     1801 ccccggagca ccgggggcac ggggtcgggc gcgcgttgat ggggctcgcg acggagttcg
     1861 cccgcgagcg gggcgccggg cacctctggc tggaggtcac caacgtcaac gcaccggcga
     1921 tccacgcgta ccggcggatg gggttcaccc tctgcggcct ggacaccgcc ctgtacgacg
     1981 gcaccgcctc ggacggcgag caggcgctct acatgagcat gccctgcccc taatcagtac
     2041 tgacaataaa aagattcttg ttttcaagaa cttgtcattt gtatagtttt tttatattgt
     2101 agttgttcta ttttaatcaa atgttagcgt gatttatatt ttttttcgcc tcgacatcat
     2161 ctgcccagat gcgaagttaa agccgaattc cgacggagcc gattttgaaa taacttcgta
     2221 tagcatacat tatacgaagt tatacggtac ccggggatcc tctagatcat gcatgagctc
     2281 actagtggat cccccgggct gcaggaattc ctcgagaagc ttgggcccgg tacctcgcga
     2341 aggccttgca ggccaaccag ataagtgaaa tctagttcca aactattttg tcatttttaa
     2401 ttttcgtatt agcttacgac gctacaccca gttcccatct attttgtcac tcttccctaa
     2461 ataatcctta aaaactccat ttccacccct cccagttccc aactattttg tccgcccaca
     2521 gcggggcatt tttcttcctg ttatgtttgg gcgctgcatt aatgaatcgg ccaacgcgcg
     2581 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga cccgctgcgc
     2641 tcggtcgttc ggctgcggcg agcggtatca gagcttatcg gccagcctcg cagagcagga
     2701 ttcccgttga gcaccgccag gtgcgaataa gggacagtga agaaggaaca cccgctcgcg
     2761 ggtgggccta cttcacctat cctgcccggc tgacgccgtt ggatacacca aggaaagtct
     2821 acacgaaccc tttggcaaaa tcctgtatat cgtgcgaaaa aggatggata taccgaaaaa
     2881 atcgctataa tgaccccgaa gcagggttat gcagcggaaa gtatacctta aggaatcccc
     2941 tgataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3001 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3061 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3121 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3181 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3241 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3301 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3361 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3421 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     3481 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3541 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3601 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3661 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3721 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3781 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3841 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     3901 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     3961 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4021 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4081 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     4141 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     4201 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     4261 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4321 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4381 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4441 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4501 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4561 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4621 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4681 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     4741 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     4801 aacctataaa aataggcgta tcacgaggcc ctttcgtc
//
