LOCUS       sequence-109990-        7431 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GmR mini-Tn7 vector, for construction of B-galactosidase protein 
            fusions, frame 1. 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 7431)
  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 7431)
  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-LacZ10
FEATURES             Location/Qualifiers
     source          1..7431
                     /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             2056..5103
                     /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(2058..2080)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     mobile_element  5177..5342
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     primer_bind     complement(5441..5458)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5612..6200)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5692..5711)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6371..7162,7163..7231))
                     /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     6994..7013
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7232..7336)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7404..7422
                     /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 caagcttgcg atcccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     2101 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     2161 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgctt tgcctggttt
     2221 ccggcaccag aagcggtgcc ggaaagctgg ctggagtgcg atcttcctga ggccgatact
     2281 gtcgtcgtcc cctcaaactg gcagatgcac ggttacgatg cgcccatcta caccaacgta
     2341 acctatccca ttacggtcaa tccgccgttt gttcccacgg agaatccgac gggttgttac
     2401 tcgctcacat ttaatgttga tgaaagctgg ctacaggaag gccagacgcg aattattttt
     2461 gatggcgtta actcggcgtt tcatctgtgg tgcaacgggc gctgggtcgg ttacggccag
     2521 gacagtcgtt tgccgtctga atttgacctg agcgcatttt tacgcgccgg agaaaaccgc
     2581 ctcgcggtga tggtgctgcg ttggagtgac ggcagttatc tggaagatca ggatatgtgg
     2641 cggatgagcg gcattttccg tgacgtctcg ttgctgcata aaccgactac acaaatcagc
     2701 gatttccatg ttgccactcg ctttaatgat gatttcagcc gcgctgtact ggaggctgaa
     2761 gttcagatgt gcggcgagtt gcgtgactac ctacgggtaa cagtttcttt atggcagggt
     2821 gaaacgcagg tcgccagcgg caccgcgcct ttcggcggtg aaattatcga tgagcgtggt
     2881 ggttatgccg atcgcgtcac actacgtctg aacgtcgaaa acccgaaact gtggagcgcc
     2941 gaaatcccga atctctatcg tgcggtggtt gaactgcaca ccgccgacgg cacgctgatt
     3001 gaagcagaag cctgcgatgt cggtttccgc gaggtgcgga ttgaaaatgg tctgctgctg
     3061 ctgaacggca agccgttgct gattcgaggc gttaaccgtc acgagcatca tcctctgcat
     3121 ggtcaggtca tggatgagca gacgatggtg caggatatcc tgctgatgaa gcagaacaac
     3181 tttaacgccg tgcgctgttc gcattatccg aaccatccgc tgtggtacac gctgtgcgac
     3241 cgctacggcc tgtatgtggt ggatgaagcc aatattgaaa cccacggcat ggtgccaatg
     3301 aatcgtctga ccgatgatcc gcgctggcta ccggcgatga gcgaacgcgt aacgcgaatg
     3361 gtgcagcgcg atcgtaatca cccgagtgtg atcatctggt cgctggggaa tgaatcaggc
     3421 cacggcgcta atcacgacgc gctgtatcgc tggatcaaat ctgtcgatcc ttcccgcccg
     3481 gtgcagtatg aaggcggcgg agccgacacc acggccaccg atattatttg cccgatgtac
     3541 gcgcgcgtgg atgaagacca gcccttcccg gctgtgccga aatggtccat caaaaaatgg
     3601 ctttcgctac ctggagagac gcgcccgctg atcctttgcg aatacgccca cgcgatgggt
     3661 aacagtcttg gcggtttcgc taaatactgg caggcgtttc gtcagtatcc ccgtttacag
     3721 ggcggcttcg tctgggactg ggtggatcag tcgctgatta aatatgatga aaacggcaac
     3781 ccgtggtcgg cttacggcgg tgattttggc gatacgccga acgatcgcca gttctgtatg
     3841 aacggtctgg tctttgccga ccgcacgccg catccagcgc tgacggaagc aaaacaccag
     3901 cagcagtttt tccagttccg tttatccggg caaaccatcg aagtgaccag cgaatacctg
     3961 ttccgtcata gcgataacga gctcctgcac tggatggtgg cgctggatgg taagccgctg
     4021 gcaagcggtg aagtgcctct ggatgtcgct ccacaaggta aacagttgat tgaactgcct
     4081 gaactaccgc agccggagag cgccgggcaa ctctggctca cagtacgcgt agtgcaaccg
     4141 aacgcgaccg catggtcaga agccgggcac atcagcgcct ggcagcagtg gcgtctggcg
     4201 gaaaacctca gtgtgacgct ccccgccgcg tcccacgcca tcccgcatct gaccaccagc
     4261 gaaatggatt tttgcatcga gctgggtaat aagcgttggc aatttaaccg ccagtcaggc
     4321 tttctttcac agatgtggat tggcgataaa aaacaactgc tgacgccgct gcgcgatcag
     4381 ttcacccgtg caccgctgga taacgacatt ggcgtaagtg aagcgacccg cattgaccct
     4441 aacgcctggg tcgaacgctg gaaggcggcg ggccattacc aggccgaagc agcgttgttg
     4501 cagtgcacgg cagatacact tgctgatgcg gtgctgatta cgaccgctca cgcgtggcag
     4561 catcagggga aaaccttatt tatcagccgg aaaacctacc ggattgatgg tagtggtcaa
     4621 atggcgatta ccgttgatgt tgaagtggcg agcgatacac cgcatccggc gcggattggc
     4681 ctgaactgcc agctggcgca ggtagcagag cgggtaaact ggctcggatt agggccgcaa
     4741 gaaaactatc ccgaccgcct tactgccgcc tgttttgacc gctgggatct gccattgtca
     4801 gacatgtata ccccgtacgt cttcccgagc gaaaacggtc tgcgctgcgg gacgcgcgaa
     4861 ttgaattatg gcccacacca gtggcgcggc gacttccagt tcaacatcag ccgctacagt
     4921 caacagcaac tgatggaaac cagccatcgc catctgctgc acgcggaaga aggcacatgg
     4981 ctgaatatcg acggtttcca tatggggatt ggtggcgacg actcctggag cccgtcagta
     5041 tcggcggatt tccagctgag cgccggtcgc taccattacc agttggtctg gtgtcaaaaa
     5101 taataataac cgggcaggcc atgtctgccc gtatttcgcg taaggaaatc cattatgtac
     5161 tatttccttg caggccaacc agataagtga aatctagttc caaactattt tgtcattttt
     5221 aattttcgta ttagcttacg acgctacacc cagttcccat ctattttgtc actcttccct
     5281 aaataatcct taaaaactcc atttccaccc ctcccagttc ccaactattt tgtccgccca
     5341 cagcggggca tttttcttcc tgttatgttt gggcgctgca ttaatgaatc ggccaacgcg
     5401 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     5461 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     5521 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     5581 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     5641 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     5701 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     5761 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     5821 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     5881 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     5941 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6001 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     6061 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6121 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     6181 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6241 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6301 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6361 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6421 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6481 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     6541 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6601 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6661 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     6721 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     6781 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     6841 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     6901 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     6961 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     7021 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     7081 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     7141 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     7201 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     7261 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     7321 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     7381 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt c
//
