LOCUS       40924_45053        4602 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pUC18T-mini-Tn7T-aad9, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4602)
  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  JOURNAL   Unpublished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 4602)
  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 4602)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4602)
  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            "Unpublished"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..4602
                     /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    955..988
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      1024..1067
                     /regulatory_class="terminator"
     CDS             complement(1066..1818)
                     /codon_start=1
                     /gene="aad9"
                     /product="spectinomycin adenyltransferase"
                     /label=aad9
                     /protein_id="ANY60780.1"
                     /translation="MNTYEQINKVKKILRKHLKNNLIGTYMFGSGVESGLKPNSDLDFL
                     VVVSEPLTDQSKEILIQKIRPISKKIGDKSNLRYIELTIIIQQEMVPWNHPPKQEFIYG
                     EWLQELYEQGYIPQKELNSDLTIMLYQAKRKNKRIYGNYDLEELLPDIPFSDVRRAIMD
                     SSEELIDNYQDDETNSILTLCRMILTMDTGKIIPKDIAGNAVAESSPLEHRERILLAVR
                     SYLGENIEWTNENVNLTINYLNNRLKKL"
     gene            complement(1066..1818)
                     /gene="aad9"
                     /label=aad9
                     /note="derived from Enterococcus faecalis"
     regulatory      1825..1830
                     /regulatory_class="ribosome_binding_site"
     regulatory      1872..1900
                     /label=chloramphenicol transacetylase gene promoter
                     /note="chloramphenicol transacetylase gene promoter"
                     /regulatory_class="promoter"
     regulatory      1872..1877
                     /regulatory_class="minus_10_signal"
     regulatory      1894..1900
                     /regulatory_class="minus_35_signal"
     misc_feature    1985..2017
                     /note="loxP; Cre recombinase site"
     misc_feature    2033..2107
                     /note="MCS; multiple cloning site"
     mobile_element  complement(2119..2284)
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     oriT            complement(2443..2551)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      complement(2783..3371)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3545..4402)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4403..4507)
                     /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 gcgtataact
      961 tcgtataatg tatgctatac gaagttattt caaaatcggc tccgtcggaa ttcataataa
     1021 aacaaaaaaa ttgaaaaaag tgtttccacc attttttcaa tttttttaca gttttttcag
     1081 gcgattattc agataattaa tggtcagatt cacattttca ttggtccatt caatattttc
     1141 gcccagatag ctgcgcacgg ccagcagaat gcgttcgcgg tgttccagcg ggctgctttc
     1201 ggccacggca ttgccggcaa tatctttcgg aataattttg ccggtatcca tggtcagaat
     1261 catgcggcac agggtcagaa tgctattggt ttcatcatcc tgataattat caatcagttc
     1321 ttcgctgcta tccataatgg cgcggcgcac atcgctaaac ggaatatccg gcagcagttc
     1381 ttccagatca taattgccat aaatgcgttt atttttgcgt ttggcctgat acagcataat
     1441 ggtcagatcg ctattcagtt ctttctgcgg aatatagccc tgttcataca gttcttgcag
     1501 ccattcgcca taaataaatt cctgtttcgg cggatgattc cacggcacca tttcctgctg
     1561 aataataatg gtcagttcaa tatagcgcag attgctttta tcgccaattt ttttgctaat
     1621 cgggcgaatt ttctgaatca gaatttcttt gctctgatcg gtcagcggtt cgctcaccac
     1681 caccagaaaa tccagatcgc tattcggttt caggccgctt tccacgccgc tgccaaacat
     1741 ataggtgcca atcagattat ttttcagatg tttgcgcaga atttttttca ctttattaat
     1801 ctgttcatag gtattcatat aaatcctcct caggataact caaaaaatac gcccggtagt
     1861 gatcttattt cattatggtg aaagttggaa cctcttacgt gccgatcaac gtctcatgaa
     1921 ttcgggatca agcctgattg ggagaaaata aaatattata ttttactgga tgaattgttt
     1981 tagtataact tcgtataatg tatgctatac gaagtagacg cgggtaccca tcatgcatga
     2041 gctcactagt ggatcccccg ggctgcagga attcctcgag aagcttgggc ccggtacctc
     2101 gcgaaggcct tgcaggccaa ccagataagt gaaatctagt tccaaactat tttgtcattt
     2161 ttaattttcg tattagctta cgacgctaca cccagttccc atctattttg tcactcttcc
     2221 ctaaataatc cttaaaaact ccatttccac ccctcccagt tcccaactat tttgtccgcc
     2281 cacagcgggg catttttctt cctgttatgt ttgggcgctg cattaatgaa tcggccaacg
     2341 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgacccgct
     2401 gcgctcggtc gttcggctgc ggcgagcggt atcagagctt atcggccagc ctcgcagagc
     2461 aggattcccg ttgagcaccg ccaggtgcga ataagggaca gtgaagaagg aacacccgct
     2521 cgcgggtggg cctacttcac ctatcctgcc cggctgacgc cgttggatac accaaggaaa
     2581 gtctacacga accctttggc aaaatcctgt atatcgtgcg aaaaaggatg gatataccga
     2641 aaaaatcgct ataatgaccc cgaagcaggg ttatgcagcg gaaagtatac cttaaggaat
     2701 cccctgataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     2761 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     2821 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     2881 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     2941 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     3001 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     3061 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     3121 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     3181 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     3241 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     3301 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     3361 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     3421 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     3481 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     3541 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     3601 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     3661 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     3721 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     3781 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     3841 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     3901 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     3961 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     4021 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     4081 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     4141 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     4201 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     4261 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     4321 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     4381 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     4441 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     4501 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     4561 cattaaccta taaaaatagg cgtatcacga ggccctttcg tc
//
