LOCUS       sequence-121762-        5374 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ZeoR mini-Tn7 vector, mobilizable, for RFP tagging bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5374)
  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 5374)
  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-dsRedExpress
FEATURES             Location/Qualifiers
     source          1..5374
                     /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(2086..2763)
                     /codon_start=1
                     /product="improved tetrameric variant of DsRed fluorescent 
                     protein"
                     /label=DsRed2
                     /note="mammalian codon-optimized"
                     /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     primer_bind     complement(2148..2171)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     2563..2583
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     RBS             2770..2781
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    2800..2816
                     /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    2833..2849
                     /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  2891..3056
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     oriT            3215..3323
                     /note="incP origin of transfer"
     rep_origin      complement(3555..4143)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3635..3654)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4314..5105,5106..5174))
                     /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     4937..4956
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5175..5279)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5347..5365
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
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 ttggtaccgg ccgccagtgt gctggaattc ggcttctaca ggaacaggtg
     2101 gtggcggccc tcggtgcgct cgtactgctc cacgatggtg tagtcctcgt tgtgggaggt
     2161 gatgtccagc ttggcgtcca cgtagtagta gccgggcagc tgcacgggct tcttggccat
     2221 gtagatggac ttgaactcca ccaggtagtg gccgccgtcc ttcagcttca gggccttgtg
     2281 ggtctcgccc ttcagcacgc cgtcgcgggg gtacaggcgc tcggtggagg cctcccagcc
     2341 catggtcttc ttctgcatca cggggccgtc ggaggggaag ttcacgccga tgaacttcac
     2401 cttgtagatg aagcagccgt cctgcaggga ggagtcctgg gtcacggtcg ccacgccgcc
     2461 gtcctcgaag ttcatcacgc gctcccactt gaagccctcg gggaaggaca gcttcttgta
     2521 gtcggggatg tcggcggggt gcttcacgta caccttggag ccgtactgga actgggggga
     2581 caggatgtcc caggcgaagg gcagggggcc gcccttggtc accttcagct tcacggtgtt
     2641 gtggccctcg taggggcggc cctcgccctc gccctcgatc tcgaactcgt ggccgttcac
     2701 ggtgccctcc atgcgcacct tgaagcgcat gaactcggtg atgacgttct cggaggaggc
     2761 catgcttaat ttctcctctt taattctaga tgtgtgaaat tgttatccgc tcacaattga
     2821 atctaagtat cattgttatc cgctcacaag tcaacactct ttttgataaa ttttctcgac
     2881 cttgcaggcc aaccagataa gtgaaatcta gttccaaact attttgtcat ttttaatttt
     2941 cgtattagct tacgacgcta cacccagttc ccatctattt tgtcactctt ccctaaataa
     3001 tccttaaaaa ctccatttcc acccctccca gttcccaact attttgtccg cccacagcgg
     3061 ggcatttttc ttcctgttat gtttgggcgc tgcattaatg aatcggccaa cgcgcgggga
     3121 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgacccg ctgcgctcgg
     3181 tcgttcggct gcggcgagcg gtatcagagc ttatcggcca gcctcgcaga gcaggattcc
     3241 cgttgagcac cgccaggtgc gaataaggga cagtgaagaa ggaacacccg ctcgcgggtg
     3301 ggcctacttc acctatcctg cccggctgac gccgttggat acaccaagga aagtctacac
     3361 gaaccctttg gcaaaatcct gtatatcgtg cgaaaaagga tggatatacc gaaaaaatcg
     3421 ctataatgac cccgaagcag ggttatgcag cggaaagtat accttaagga atcccctgat
     3481 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     3541 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3601 tcgagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     3661 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3721 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3781 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3841 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3901 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     3961 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     4021 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     4081 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4141 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     4201 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     4261 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     4321 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     4381 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     4441 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     4501 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     4561 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     4621 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     4681 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     4741 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     4801 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     4861 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     4921 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     4981 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     5041 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     5101 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     5161 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     5221 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     5281 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     5341 tataaaaata ggcgtatcac gaggcccttt cgtc
//
