LOCUS       sequence-121763-        5578 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ZeoR mini-Tn7 vector, mobilizable, for YFP tagging bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5578)
  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 5578)
  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-eyfp
FEATURES             Location/Qualifiers
     source          1..5578
                     /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(967..1014)
                     /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(1097..1471)
                     /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(1490..1537)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     protein_bind    complement(1648..1695)
                     /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(1749..1835)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(1938..2032)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(2055..2768,2769..2771,2772..2774))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MLSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2100..2121)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2449..2468
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2708..2729
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     RBS             2781..2792
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    2811..2827
                     /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    2844..2860
                     /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  3095..3260
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     oriT            3419..3527
                     /note="incP origin of transfer"
     rep_origin      complement(3759..4347)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3839..3858)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4518..5309,5310..5378))
                     /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     5141..5160
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5379..5483)
                     /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 ggcccggacc gatcgaatta gcttcaaaag
      961 cgctctgaag ttcctatact ttctagagaa taggaacttc ggaataggaa cttcaagatc
     1021 ccctgattcc ctttgtcaac agcaatggat ttatctctga tgttacattg cacaagataa
     1081 aaatatatca tcatgatcag tcctgctcct cggccacgaa gtgcacgcag ttgccggccg
     1141 ggtcgcgcag ggcgaactcc cgcccccacg gctgctcgcc gatctcggtc atggccggcc
     1201 cggaggcgtc ccggaagttc gtggacacga cctccgacca ctcggcgtac agctcgtcca
     1261 ggccgcgcac ccacacccag gccagggtgt tgtccggcac cacctggtcc tggaccgcgc
     1321 tgatgaacag ggtcacgtcg tcccggacca caccggcgaa gtcgtcctcc acgaagtccc
     1381 gggagaaccc gagccggtcg gtccagaact cgaccgctcc ggcgacgtcg cgcgcggtga
     1441 gcaccggaac ggcactggtc aacttggcca tggtttagtt cctcaccttg tcgtattata
     1501 ctatgccgat atactatgcc gatgattaat tgtcaacacg tgctgatcat gaccaaaatc
     1561 ccttaacgtg agttacgcgt cgaagccgaa tatctagaat tattccattg agtaagtttt
     1621 taagcacatc agcttcaaaa gcgctctgaa gttcctatac tttctagaga ataggaactt
     1681 cggaataggt acttcaagat ccccaattcg atcatgcatg agctcgggcc gcaagctcct
     1741 agcggcggat ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag
     1801 gcccagtctt tcgactgagc ctttcgtttt atttgatgcc tcaagctaga gagtcattac
     1861 cccaggcgtt taagggcacc aataactgcc ttaaaaaaat tacgccccgc cctgccactc
     1921 atcgcagtct agcttggatt ctcaccaata aaaaacgccc ggcggcaacc gagcgttctg
     1981 aacaaatcca gatggagttc tgaggtcatt actggatcta tcaacaggag tccaagctca
     2041 gctaattaag cttattactt gtacagctcg tccatgccga gagtgatccc ggcggcggtc
     2101 acgaactcca gcaggaccat gtgatcgcgc ttctcgttgg ggtctttgct cagggcggac
     2161 tggtagctca ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat gggggtgttc
     2221 tgctggtagt ggtcggcgag ctgcacgctg ccgtcctcga tgttgtggcg gatcttgaag
     2281 ttcaccttga tgccgttctt ctgcttgtcg gccatgatat agacgttgtg gctgttgtag
     2341 ttgtactcca gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat gcccttcagc
     2401 tcgatgcggt tcaccagggt gtcgccctcg aacttcacct cggcgcgggt cttgtagttg
     2461 ccgtcgtcct tgaagaagat ggtgcgctcc tggacgtagc cttcgggcat ggcggacttg
     2521 aagaagtcgt gctgcttcat gtggtcgggg tagcgggcga agcactgcag gccgtagccg
     2581 aaggtggtca cgagggtggg ccagggcacg ggcagcttgc cggtggtgca gatgaacttc
     2641 agggtcagct tgccgtaggt ggcatcgccc tcgccctcgc cggacacgct gaacttgtgg
     2701 ccgtttacgt cgccgtccag ctcgaccagg atgggcacca ccccggtgaa cagctcctcg
     2761 cccttgctca gcatgcttaa tttctcctct ttaattctag atgtgtgaaa ttgttatccg
     2821 ctcacaattg aatctaagta tcattgttat ccgctcacaa gtcaacactc tttttgataa
     2881 attttctcga ggtgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     2941 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     3001 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     3061 tgataaatgg tacctcgcga aggccttgca ggccaaccag ataagtgaaa tctagttcca
     3121 aactattttg tcatttttaa ttttcgtatt agcttacgac gctacaccca gttcccatct
     3181 attttgtcac tcttccctaa ataatcctta aaaactccat ttccacccct cccagttccc
     3241 aactattttg tccgcccaca gcggggcatt tttcttcctg ttatgtttgg gcgctgcatt
     3301 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     3361 cgctcactga cccgctgcgc tcggtcgttc ggctgcggcg agcggtatca gagcttatcg
     3421 gccagcctcg cagagcagga ttcccgttga gcaccgccag gtgcgaataa gggacagtga
     3481 agaaggaaca cccgctcgcg ggtgggccta cttcacctat cctgcccggc tgacgccgtt
     3541 ggatacacca aggaaagtct acacgaaccc tttggcaaaa tcctgtatat cgtgcgaaaa
     3601 aggatggata taccgaaaaa atcgctataa tgaccccgaa gcagggttat gcagcggaaa
     3661 gtatacctta aggaatcccc tgataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     3721 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3781 gacgagcatc acaaaaatcg acgctcgagt cagaggtggc gaaacccgac aggactataa
     3841 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3901 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3961 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4021 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4081 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4141 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     4201 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4261 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     4321 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     4381 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     4441 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     4501 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     4561 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     4621 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     4681 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     4741 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     4801 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     4861 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     4921 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     4981 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     5041 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     5101 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     5161 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     5221 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     5281 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     5341 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     5401 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     5461 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     5521 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtc
//
