LOCUS       40924_45003        5874 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pUC18-mini-Tn7T-Tp-Gateway, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5874)
  AUTHORS   Choi K-H., Gaynor JB, White KG, Karkhoff-Schweizer RR, Schweizer HP.
  TITLE     A Tn7-based universal bacterial cloning and expression system
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5874)
  AUTHORS   Choi K-H., Gaynor JB, White KG, Karkhoff-Schweizer RR, Schweizer HP.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-AUG-2004) Microbiology, Immunology and Pathology, 
            Colorado State University, Fort Collins, CO 80523, USA
REFERENCE   3  (bases 1 to 5874)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5874)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (27-AUG-2004) Microbiology, Immunology and Pathology, Colorado State
            University, Fort Collins, CO 80523, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5874
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   382..580
                     /label=Tn7R, right end of Tn7
                     /note="Tn7R, 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    964..1011
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(1076..1309)
                     /codon_start=1
                     /label=TpR
                     /note="E. coli plasmid-associated dihydrofolate reductase"
                     /translation="MGQSSDEANAPVAGQFALPLSATFGLGDRVRKKSGAAWQGQVVGW
                     YCTKLTPEGYAVESESHPGSVQIYPVAALERVA"
     promoter        complement(1636..1664)
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     misc_feature    1716..1763
                     /label=FRT, Flp recombinase target site
                     /note="FRT, Flp recombinase target site"
     protein_bind    1716..1763
                     /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)."
     protein_bind    1885..2009
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(2053..2355)
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     CDS             complement(2700..3356)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(3410..3440)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     protein_bind    complement(3465..3589)
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     mobile_element  complement(3620..3785)
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      complement(4055..4643)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4817..5674)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5675..5779)
                     /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 atcgagctcg aattggggat
      961 cttgaagtac ctattccgaa gttcctattc tctagaaagt ataggaactt cagagcgctt
     1021 ttgaagctga tgtgcttaaa aacttactca atggaataat tctagataat tcttaggcca
     1081 cacgttcaag tgcagccaca ggataaattt gcactgagcc tgggtgggat tcggactcga
     1141 ccgcatagcc ttcaggagtg agttttgtgc aataccaacc gacgacttga ccctgccaag
     1201 cggcaccaga tttcttgcgt acgcgatccc ctaagccaaa ggtggcactc aggggaagcg
     1261 caaactgccc tgcaacggga gcgttggctt catcgctact ttgacccatg tcgaatcctt
     1321 cttgtgaatc tattatggcg acaagcaaat cgagctctga ctgcctaccc cacaacaact
     1381 actagaaagc accagcacaa cggctgccta actttgtttt agggcgacta ccctgctgcg
     1441 taacatcgtt gctgttccat aacatcaaac atcgacccac ggcgtaacgc gcttagctag
     1501 cttggatgcc cgaggctaga ctgtacaaaa aaacagtcat aacaagccat gaaaaccgcc
     1561 actgcgccgt taccaccgct gcgttcggtc aaggttctgg accagttgcg tgagcgcata
     1621 cgctacttgc attacagttt acgaaccgaa caggcttatg tcaactgggt tcgtgaatta
     1681 tccattgctg ttgacaaagg gaatcagggg atcttgaagt tcctattccg aagttcctat
     1741 tctctagaaa gtataggaac ttcagagcgc ttttgaagct aattcgagct cgatcatgca
     1801 tgagctcact agtggatccc ccgggctgca ggaattcctc gagaagcttg catgcctgca
     1861 ggtcgactct agaggatccc catcaccact ttgtacaaga aagctgaacg agaaacgtaa
     1921 aatgatataa atatcaatat attaaattag attttgcata aaaaacagac tacataatac
     1981 tgtaaaacac aacatatcca gtcactatgg tcgacctgca gactggctgt gtataacgga
     2041 gcctgacatt tatattcccc agaacatcag gttaatggcg tttttgatgt cattttcgcg
     2101 gtggctgaga tcagccactt cttccccgat aacggagacc ggcacactgg ccatatcggt
     2161 ggtcatcatg cgccagcttt catccccgat atgcaccacc gggtaaagtt cacgggagac
     2221 tttatctgac agcagacgtg cactggccag ggggatcacc atccgtcgcc cgggcgtgtc
     2281 aataatatca ctctgtacat ccacaaacag acgataacgg ctctctcttt tataggtgta
     2341 aaccttaaac tgcatttcac cagtccctgt tctcgtcagc aaaagagccg ttcatttcaa
     2401 taaaccgggc gacctcagcc atcccttcct gattttccgc tttccagcgt tcggcacgca
     2461 gacgacgggc ttcattctgc atggttgtgc ttaccagacc ggagatattg acatcatata
     2521 tgccttgagc aactgatagc tgtcgctgtc aactgtcact gtaatacgct gcttcatagc
     2581 acacctcttt ttgacatact tcgggtatac atatcagtat atattcttat accgcaaaaa
     2641 tcagcgcgca aatacgcata ctgttatctg gcttttagta agccggatcc acgcgtttac
     2701 gccccgccct gccactcatc gcagtactgt tgtaattcat taagcattct gccgacatgg
     2761 aagccatcac agacggcatg atgaacctga atcgccagcg gcatcagcac cttgtcgcct
     2821 tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga agttgtccat attggccacg
     2881 tttaaatcaa aactggtgaa actcacccag ggattggctg agacgaaaaa catattctca
     2941 ataaaccctt tagggaaata ggccaggttt tcaccgtaac acgccacatc ttgcgaatat
     3001 atgtgtagaa actgccggaa atcgtcgtgg tattcactcc agagcgatga aaacgtttca
     3061 gtttgctcat ggaaaacggt gtaacaaggg tgaacactat cccatatcac cagctcaccg
     3121 tctttcattg ccatacggaa ttccggatga gcattcatca ggcgggcaag aatgtgaata
     3181 aaggccggat aaaacttgtg cttatttttc tttacggtct ttaaaaaggc cgtaatatcc
     3241 agctgaacgg tctggttata ggtacattga gcaactgact gaaatgcctc aaaatgttct
     3301 ttacgatgcc attgggatat atcaacggtg gtatatccag tgattttttt ctccatttta
     3361 gcttccttag ctcctgaaaa tctcgccgga tcctaactca aaatccacac attatacgag
     3421 ccggaagcat aaagtgtaaa gcctggggtg cctaatgcgg ccgccatagt gactggatat
     3481 gttgtgtttt acagtattat gtagtctgtt ttttatgcaa aatctaattt aatatattga
     3541 tatttatatc attttacgtt tctcgttcag cttttttgta caaacttgtg atgggtacct
     3601 cgcgaaggcc ttgcaggcca accagataag tgaaatctag ttccaaacta ttttgtcatt
     3661 tttaattttc gtattagctt acgacgctac acccagttcc catctatttt gtcactcttc
     3721 cctaaataat ccttaaaaac tccatttcca cccctcccag ttcccaacta ttttgtccgc
     3781 ccacagcggg gcatttttct tcctgttatg tttgggcgct gcattaatga atcggccaac
     3841 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     3901 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     3961 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     4021 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4081 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4141 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4201 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4261 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4321 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4381 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     4441 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     4501 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     4561 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     4621 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     4681 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     4741 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     4801 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     4861 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     4921 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     4981 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     5041 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     5101 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     5161 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     5221 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     5281 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     5341 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     5401 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     5461 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     5521 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     5581 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     5641 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     5701 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     5761 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     5821 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtc
//
