LOCUS       sequence-121764-        5577 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ZeoR mini-Tn7 vector, mobilizable, for CFP tagging bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5577)
  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 5577)
  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-Zeo-ecfp
FEATURES             Location/Qualifiers
     source          1..5577
                     /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    complement(966..1013)
                     /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(1096..1470)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     promoter        complement(1489..1536)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     protein_bind    complement(1647..1694)
                     /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(1748..1834)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(1937..2031)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(2054..2767,2768..2770,2771..2773))
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MLSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2099..2120)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2448..2467
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2707..2728
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     RBS             2780..2791
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    2810..2826
                     /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    2843..2859
                     /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  3094..3259
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     oriT            3418..3526
                     /note="incP origin of transfer"
     rep_origin      complement(3758..4346)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3838..3857)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4517..5308,5309..5377))
                     /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     5140..5159
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5378..5482)
                     /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 atcgaattag cttcaaaagc
      961 gctctgaagt tcctatactt tctagagaat aggaacttcg gaataggaac ttcaagatcc
     1021 cctgattccc tttgtcaaca gcaatggatt tatctctgat gttacattgc acaagataaa
     1081 aatatatcat catgatcagt cctgctcctc ggccacgaag tgcacgcagt tgccggccgg
     1141 gtcgcgcagg gcgaactccc gcccccacgg ctgctcgccg atctcggtca tggccggccc
     1201 ggaggcgtcc cggaagttcg tggacacgac ctccgaccac tcggcgtaca gctcgtccag
     1261 gccgcgcacc cacacccagg ccagggtgtt gtccggcacc acctggtcct ggaccgcgct
     1321 gatgaacagg gtcacgtcgt cccggaccac accggcgaag tcgtcctcca cgaagtcccg
     1381 ggagaacccg agccggtcgg tccagaactc gaccgctccg gcgacgtcgc gcgcggtgag
     1441 caccggaacg gcactggtca acttggccat ggtttagttc ctcaccttgt cgtattatac
     1501 tatgccgata tactatgccg atgattaatt gtcaacacgt gctgatcatg accaaaatcc
     1561 cttaacgtga gttacgcgtc gaagccgaat atctagaatt attccattga gtaagttttt
     1621 aagcacatca gcttcaaaag cgctctgaag ttcctatact ttctagagaa taggaacttc
     1681 ggaataggta cttcaagatc cccaattcga tcatgcatga gctcgggccg caagctccta
     1741 gcggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg
     1801 cccagtcttt cgactgagcc tttcgtttta tttgatgcct caagctagag agtcattacc
     1861 ccaggcgttt aagggcacca ataactgcct taaaaaaatt acgccccgcc ctgccactca
     1921 tcgcagtcta gcttggattc tcaccaataa aaaacgcccg gcggcaaccg agcgttctga
     1981 acaaatccag atggagttct gaggtcatta ctggatctat caacaggagt ccaagctcag
     2041 ctaattaagc ttattacttg tacagctcgt ccatgccgag agtgatcccg gcggcggtca
     2101 cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttgctc agggcggact
     2161 gggtgctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg ggggtgttct
     2221 gctggtagtg gtcggcgagc tgcacgctgc cgtcctcgat gttgtggcgg atcttgaagt
     2281 tggccttgat gccgttcttc tgcttgtcgg cggtgatata gacgttgtgg ctgatgtagt
     2341 tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg cccttcagct
     2401 cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc
     2461 cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga
     2521 agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa gcactgcacg ccccaggtca
     2581 gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag atgaacttca
     2641 gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg aacttgtggc
     2701 cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac agctcctcgc
     2761 ccttgctcag catgcttaat ttctcctctt taattctaga tgtgtgaaat tgttatccgc
     2821 tcacaattga atctaagtat cattgttatc cgctcacaag tcaacactct ttttgataaa
     2881 ttttctcgag gtgaagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     2941 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3001 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     3061 gataaatggt acctcgcgaa ggccttgcag gccaaccaga taagtgaaat ctagttccaa
     3121 actattttgt catttttaat tttcgtatta gcttacgacg ctacacccag ttcccatcta
     3181 ttttgtcact cttccctaaa taatccttaa aaactccatt tccacccctc ccagttccca
     3241 actattttgt ccgcccacag cggggcattt ttcttcctgt tatgtttggg cgctgcatta
     3301 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     3361 gctcactgac ccgctgcgct cggtcgttcg gctgcggcga gcggtatcag agcttatcgg
     3421 ccagcctcgc agagcaggat tcccgttgag caccgccagg tgcgaataag ggacagtgaa
     3481 gaaggaacac ccgctcgcgg gtgggcctac ttcacctatc ctgcccggct gacgccgttg
     3541 gatacaccaa ggaaagtcta cacgaaccct ttggcaaaat cctgtatatc gtgcgaaaaa
     3601 ggatggatat accgaaaaaa tcgctataat gaccccgaag cagggttatg cagcggaaag
     3661 tataccttaa ggaatcccct gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     3721 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     3781 acgagcatca caaaaatcga cgctcgagtc agaggtggcg aaacccgaca ggactataaa
     3841 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3901 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     3961 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4021 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4081 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4141 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     4201 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4261 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4321 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4381 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     4441 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     4501 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     4561 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     4621 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     4681 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     4741 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     4801 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     4861 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     4921 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     4981 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     5041 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     5101 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     5161 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     5221 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     5281 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     5341 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     5401 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     5461 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     5521 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtc
//
