LOCUS       sequence-109982-        5707 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TpR mini-Tn7 vector, mobilizable, for CFP tagging bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5707)
  AUTHORS   Choi KH, Schweizer HP
  TITLE     mini-Tn7 insertion in bacteria with single attTn7 sites: example 
            Pseudomonas aeruginosa.
  JOURNAL   Nat Protoc. 2006;1(1):153-61.
  PUBMED    17406227
REFERENCE   2  (bases 1 to 5707)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Protoc."; date: "2006"; volume: "1(1)"; pages: "153-61"
COMMENT     Sequence Label: pUC18T-mini-Tn7T-Tp-ecfp
FEATURES             Location/Qualifiers
     source          1..5707
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     584..600
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     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
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    963..1010
                     /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(1072..1308)
                     /codon_start=1
                     /gene="dhfrIIb from plasmid R388"
                     /product="E. coli plasmid-associated dihydrofolate 
                     reductase"
                     /label=TpR
                     /note="confers resistance to trimethoprim"
                     /translation="MGQSSDEANAPVAGQFALPLSATFGLGDRVRKKSGAAWQGQVVGW
                     YCTKLTPEGYAVESESHPGSVQIYPVAALERVA"
     promoter        complement(join(1633..1638,1639..1655,1656..1661))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    1713..1760
                     /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)."
     terminator      complement(1892..1978)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(2081..2175)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(2198..2911,2912..2914,2915..2917))
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MLSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2243..2264)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2592..2611
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2851..2872
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     RBS             2924..2935
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    2954..2970
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     protein_bind    2987..3003
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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  3224..3389
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     oriT            3548..3656
                     /note="incP origin of transfer"
     rep_origin      complement(3888..4476)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3968..3987)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4647..5438,5439..5507))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5270..5289
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5508..5612)
                     /gene="bla"
                     /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 atgagctcga attggggatc
      961 ttgaagtacc tattccgaag ttcctattct ctagaaagta taggaacttc agagcgcttt
     1021 tgaagctgat gtgcttaaaa acttactcaa tggaataatt ctagataatt cttaggccac
     1081 acgttcaagt gcagccacag gataaatttg cactgagcct gggtgggatt cggactcgac
     1141 cgcatagcct tcaggagtga gttttgtgca ataccaaccg acgacttgac cctgccaagc
     1201 ggcaccagat ttcttgcgta cgcgatcccc taagccaaag gtggcactca ggggaagcgc
     1261 aaactgccct gcaacgggag cgttggcttc atcgctactt tgacccatgt cgaatccttc
     1321 ttgtgaatct attatggcga caagcaaatc gagctctgac tgcctacccc acaacaacta
     1381 tcagaaagca ccagcacaac ggctgcctaa ctttgtttta gggcgactgc cctgctgcgt
     1441 aacatcgttg ctgctccata acatcaaaca tcgacccacg gcgtaacgcg cttgctgctt
     1501 ggatgcccga ggctagactg tacaaaaaaa cagtcataac aagccatgaa aaccgccact
     1561 gcgccgttac caccgctgcg ttcggtcaag gttctggacc agttgcgtga gcgcatacgc
     1621 tacttgcatt acagtttacg aaccgaacag gcttatgtca actgggttcg tgaattatcc
     1681 attgctgttg acaaagggaa tcaggggatc ttgaagttcc tattccgaag ttcctattct
     1741 ctagaaagta taggaacttc agagcgcttt tgaagctaat tcgagctcga tcatgcatga
     1801 gctcactagt ggatcccccg ggctgcagga attcctcgag aagcttgggc ccggtacctc
     1861 gcgaaggcgg gccgcaagct cttagcggcg gatttgtcct actcaggaga gcgttcaccg
     1921 acaaacaaca gataaaacga aaggcccagt ctttcgactg agcctttcgt tttatttgat
     1981 gcctcaagct agagagtcat taccccaggc gtttaagggc accaataact gccttaaaaa
     2041 aattacgccc cgccctgcca ctcatcgcag tctagcttgg attctcacca ataaaaaacg
     2101 cccggcggca accgagcgtt ctgaacaaat ccagatggag ttctgaggtc attactggat
     2161 ctatcaacag gagtccaagc tcagctaatt aagcttatta cttgtacagc tcgtccatgc
     2221 cgagagtgat cccggcggcg gtcacgaact ccagcaggac catgtgatcg cgcttctcgt
     2281 tggggtcttt gctcagggcg gactgggtgc tcaggtagtg gttgtcgggc agcagcacgg
     2341 ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc gagctgcacg ctgccgtcct
     2401 cgatgttgtg gcggatcttg aagttggcct tgatgccgtt cttctgcttg tcggcggtga
     2461 tatagacgtt gtggctgatg tagttgtact ccagcttgtg ccccaggatg ttgccgtcct
     2521 ccttgaagtc gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca
     2581 cctcggcgcg ggtcttgtag ttgccgtcgt ccttgaagaa gatggtgcgc tcctggacgt
     2641 agccttcggg catggcggac ttgaagaagt cgtgctgctt catgtggtcg gggtagcggc
     2701 tgaagcactg cacgccccag gtcagggtgg tcacgagggt gggccagggc acgggcagct
     2761 tgccggtggt gcagatgaac ttcagggtca gcttgccgta ggtggcatcg ccctcgccct
     2821 cgccggacac gctgaacttg tggccgttta cgtcgccgtc cagctcgacc aggatgggca
     2881 ccaccccggt gaacagctcc tcgcccttgc tcagcatgct taatttctcc tctttaattc
     2941 tagatgtgtg aaattgttat ccgctcacaa ttgaatctaa gtatcattgt tatccgctca
     3001 caagtcaaca ctctttttga taaattttct cgaggtgaag acgaaagggc ctcgtgatac
     3061 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     3121 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     3181 atccgctcat gagacaataa ccctgataaa tgccttgcag gccaaccaga taagtgaaat
     3241 ctagttccaa actattttgt catttttaat tttcgtatta gcttacgacg ctacacccag
     3301 ttcccatcta ttttgtcact cttccctaaa taatccttaa aaactccatt tccacccctc
     3361 ccagttccca actattttgt ccgcccacag cggggcattt ttcttcctgt tatgtttggg
     3421 cgctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     3481 ccgcttcctc gctcactgac ccgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     3541 agcttatcgg ccagcctcgc agagcaggat tcccgttgag caccgccagg tgcgaataag
     3601 ggacagtgaa gaaggaacac ccgctcgcgg gtgggcctac ttcacctatc ctgcccggct
     3661 gacgccgttg gatacaccaa ggaaagtcta cacgaaccct ttggcaaaat cctgtatatc
     3721 gtgcgaaaaa ggatggatat accgaaaaaa tcgctataat gaccccgaag cagggttatg
     3781 cagcggaaag tataccttaa ggaatcccct gataacgcag gaaagaacat gtgagcaaaa
     3841 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     3901 cgcccccctg acgagcatca caaaaatcga cgctcgagtc agaggtggcg aaacccgaca
     3961 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4021 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4081 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4141 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4201 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4261 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     4321 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     4381 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     4441 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     4501 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     4561 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     4621 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     4681 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     4741 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     4801 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     4861 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     4921 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     4981 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     5041 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     5101 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     5161 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     5221 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     5281 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     5341 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     5401 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     5461 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     5521 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     5581 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     5641 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     5701 tttcgtc
//
