LOCUS       sequence-109988-        7430 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GmR mini-Tn7 vector, for construction of B-galactosidase protein 
            fusions, frame 3. See Depositor Comments below for information 
            regarding the lacZ sequence..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7430)
  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 7430)
  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: pUC18-mini-Tn7T-Gm-LacZ30
FEATURES             Location/Qualifiers
     source          1..7430
                     /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    958..1005
                     /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(1135..1668)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(1857..1862,1863..1879,1880..1885))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    1926..1973
                     /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             2055..5102
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-galactosidase"
                     /label=lacZ
                     /translation="VVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRS
                     LNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNVTYPITVNPP
                     FVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYGQDSRLPSEF
                     DLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQISDFHVATR
                     FNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIIDERGGYADRV
                     TLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIENGLLLLNGKP
                     LLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLWYTLCDRYGL
                     YVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWSLGNESGHGA
                     NHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVPKWSIKKWLS
                     LPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSLIKYDENGNP
                     WSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQTIEVTSEYL
                     FRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQLWLTVRVVQ
                     PNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELGNKRWQFNRQ
                     SGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWKAAGHYQAEA
                     ALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDVEVASDTPHP
                     ARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPYVFPSENGLR
                     CGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDGFHMGIGGDD
                     SWSPSVSADFQLSAGRYHYQLVWCQK"
     primer_bind     complement(2057..2079)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     mobile_element  5176..5341
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     primer_bind     complement(5440..5457)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5611..6199)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5691..5710)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6370..7161,7162..7230))
                     /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     6993..7012
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7231..7335)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7403..7421
                     /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 atcgaattgg ggatcttgaa
      961 gtacctattc cgaagttcct attctctaga aagtatagga acttcagagc gcttttgaag
     1021 ctgatgtgct taaaaactta ctcaatggaa ttctagatcg aattggccgc ggcgttgtga
     1081 caatttaccg aacaactccg cggccgggaa gccgatctcg gcttgaacga attgttaggt
     1141 ggcggtactt gggtcgatat caaagtgcat cacttcttcc cgtatgccca actttgtata
     1201 gagagccact gcgggatcgt caccgtaatc tgcttgcacg tagatcacat aagcaccaag
     1261 cgcgttggcc tcatgcttga ggagattgat gagcgcggtg gcaatgccct gcctccggtg
     1321 ctcgccggag actgcgagat catagatata gatctcacta cgcggctgct caaacttggg
     1381 cagaacgtaa gccgcgagag cgccaacaac cgcttcttgg tcgaaggcag caagcgcgat
     1441 gaatgtctta ctacggagca agttcccgag gtaatcggag tccggctgat gttgggagta
     1501 ggtggctacg tctccgaact cacgaccgaa aagatcaaga gcagcccgca tggatttgac
     1561 ttggtcaggg ccgagcctac atgtgcgaat gatgcccata cttgagccac ctaactttgt
     1621 tttagggcga ctgccctgct gcgtaacatc gttgctgctg cgtaacatcg ttgctgctcc
     1681 ataacatcaa acatcgaccc acggcgtaac gcgcttgctg cttggatgcc cgaggcatag
     1741 actgtacaaa aaaacagtca taacaagcca tgaaaaccgc cactgcgccg ttaccaccgc
     1801 tgcgttcggt caaggttctg gaccagttgc gtgagcgcat acgctacttg cattacagtt
     1861 tacgaaccga acaggcttat gtcaattcga tccattgctg ttgacaaagg gaatcagggg
     1921 atcttgaagt tcctattccg aagttcctat tctctagaaa gtataggaac ttcagagcgc
     1981 ttttgaagct aattcgatca tgcatgagct cgaattcccg gggatccgtc gacctgcagc
     2041 caagcttcga tcccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
     2101 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
     2161 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcttt gcctggtttc
     2221 cggcaccaga agcggtgccg gaaagctggc tggagtgcga tcttcctgag gccgatactg
     2281 tcgtcgtccc ctcaaactgg cagatgcacg gttacgatgc gcccatctac accaacgtaa
     2341 cctatcccat tacggtcaat ccgccgtttg ttcccacgga gaatccgacg ggttgttact
     2401 cgctcacatt taatgttgat gaaagctggc tacaggaagg ccagacgcga attatttttg
     2461 atggcgttaa ctcggcgttt catctgtggt gcaacgggcg ctgggtcggt tacggccagg
     2521 acagtcgttt gccgtctgaa tttgacctga gcgcattttt acgcgccgga gaaaaccgcc
     2581 tcgcggtgat ggtgctgcgt tggagtgacg gcagttatct ggaagatcag gatatgtggc
     2641 ggatgagcgg cattttccgt gacgtctcgt tgctgcataa accgactaca caaatcagcg
     2701 atttccatgt tgccactcgc tttaatgatg atttcagccg cgctgtactg gaggctgaag
     2761 ttcagatgtg cggcgagttg cgtgactacc tacgggtaac agtttcttta tggcagggtg
     2821 aaacgcaggt cgccagcggc accgcgcctt tcggcggtga aattatcgat gagcgtggtg
     2881 gttatgccga tcgcgtcaca ctacgtctga acgtcgaaaa cccgaaactg tggagcgccg
     2941 aaatcccgaa tctctatcgt gcggtggttg aactgcacac cgccgacggc acgctgattg
     3001 aagcagaagc ctgcgatgtc ggtttccgcg aggtgcggat tgaaaatggt ctgctgctgc
     3061 tgaacggcaa gccgttgctg attcgaggcg ttaaccgtca cgagcatcat cctctgcatg
     3121 gtcaggtcat ggatgagcag acgatggtgc aggatatcct gctgatgaag cagaacaact
     3181 ttaacgccgt gcgctgttcg cattatccga accatccgct gtggtacacg ctgtgcgacc
     3241 gctacggcct gtatgtggtg gatgaagcca atattgaaac ccacggcatg gtgccaatga
     3301 atcgtctgac cgatgatccg cgctggctac cggcgatgag cgaacgcgta acgcgaatgg
     3361 tgcagcgcga tcgtaatcac ccgagtgtga tcatctggtc gctggggaat gaatcaggcc
     3421 acggcgctaa tcacgacgcg ctgtatcgct ggatcaaatc tgtcgatcct tcccgcccgg
     3481 tgcagtatga aggcggcgga gccgacacca cggccaccga tattatttgc ccgatgtacg
     3541 cgcgcgtgga tgaagaccag cccttcccgg ctgtgccgaa atggtccatc aaaaaatggc
     3601 tttcgctacc tggagagacg cgcccgctga tcctttgcga atacgcccac gcgatgggta
     3661 acagtcttgg cggtttcgct aaatactggc aggcgtttcg tcagtatccc cgtttacagg
     3721 gcggcttcgt ctgggactgg gtggatcagt cgctgattaa atatgatgaa aacggcaacc
     3781 cgtggtcggc ttacggcggt gattttggcg atacgccgaa cgatcgccag ttctgtatga
     3841 acggtctggt ctttgccgac cgcacgccgc atccagcgct gacggaagca aaacaccagc
     3901 agcagttttt ccagttccgt ttatccgggc aaaccatcga agtgaccagc gaatacctgt
     3961 tccgtcatag cgataacgag ctcctgcact ggatggtggc gctggatggt aagccgctgg
     4021 caagcggtga agtgcctctg gatgtcgctc cacaaggtaa acagttgatt gaactgcctg
     4081 aactaccgca gccggagagc gccgggcaac tctggctcac agtacgcgta gtgcaaccga
     4141 acgcgaccgc atggtcagaa gccgggcaca tcagcgcctg gcagcagtgg cgtctggcgg
     4201 aaaacctcag tgtgacgctc cccgccgcgt cccacgccat cccgcatctg accaccagcg
     4261 aaatggattt ttgcatcgag ctgggtaata agcgttggca atttaaccgc cagtcaggct
     4321 ttctttcaca gatgtggatt ggcgataaaa aacaactgct gacgccgctg cgcgatcagt
     4381 tcacccgtgc accgctggat aacgacattg gcgtaagtga agcgacccgc attgacccta
     4441 acgcctgggt cgaacgctgg aaggcggcgg gccattacca ggccgaagca gcgttgttgc
     4501 agtgcacggc agatacactt gctgatgcgg tgctgattac gaccgctcac gcgtggcagc
     4561 atcaggggaa aaccttattt atcagccgga aaacctaccg gattgatggt agtggtcaaa
     4621 tggcgattac cgttgatgtt gaagtggcga gcgatacacc gcatccggcg cggattggcc
     4681 tgaactgcca gctggcgcag gtagcagagc gggtaaactg gctcggatta gggccgcaag
     4741 aaaactatcc cgaccgcctt actgccgcct gttttgaccg ctgggatctg ccattgtcag
     4801 acatgtatac cccgtacgtc ttcccgagcg aaaacggtct gcgctgcggg acgcgcgaat
     4861 tgaattatgg cccacaccag tggcgcggcg acttccagtt caacatcagc cgctacagtc
     4921 aacagcaact gatggaaacc agccatcgcc atctgctgca cgcggaagaa ggcacatggc
     4981 tgaatatcga cggtttccat atggggattg gtggcgacga ctcctggagc ccgtcagtat
     5041 cggcggattt ccagctgagc gccggtcgct accattacca gttggtctgg tgtcaaaaat
     5101 aataataacc gggcaggcca tgtctgcccg tatttcgcgt aaggaaatcc attatgtact
     5161 atttccttgc aggccaacca gataagtgaa atctagttcc aaactatttt gtcattttta
     5221 attttcgtat tagcttacga cgctacaccc agttcccatc tattttgtca ctcttcccta
     5281 aataatcctt aaaaactcca tttccacccc tcccagttcc caactatttt gtccgcccac
     5341 agcggggcat ttttcttcct gttatgtttg ggcgctgcat taatgaatcg gccaacgcgc
     5401 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5461 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5521 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5581 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5641 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5701 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5761 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5821 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5881 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5941 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6001 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     6061 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6121 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     6181 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6241 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6301 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     6361 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6421 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6481 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6541 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6601 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6661 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6721 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6781 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6841 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6901 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6961 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     7021 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     7081 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     7141 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7201 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7261 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     7321 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     7381 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
//
