LOCUS       sequence-109989-        7429 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GmR mini-Tn7 vector, for construction of B-galactosidase protein 
            fusions, frame 2. 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 7429)
  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 7429)
  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-LacZ20
FEATURES             Location/Qualifiers
     source          1..7429
                     /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             2054..5101
                     /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(2056..2078)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     mobile_element  5175..5340
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     primer_bind     complement(5439..5456)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5610..6198)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5690..5709)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6369..7160,7161..7229))
                     /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     6992..7011
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7230..7334)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7402..7420
                     /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 caagcttgct cccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     2101 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     2161 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgctttg cctggtttcc
     2221 ggcaccagaa gcggtgccgg aaagctggct ggagtgcgat cttcctgagg ccgatactgt
     2281 cgtcgtcccc tcaaactggc agatgcacgg ttacgatgcg cccatctaca ccaacgtaac
     2341 ctatcccatt acggtcaatc cgccgtttgt tcccacggag aatccgacgg gttgttactc
     2401 gctcacattt aatgttgatg aaagctggct acaggaaggc cagacgcgaa ttatttttga
     2461 tggcgttaac tcggcgtttc atctgtggtg caacgggcgc tgggtcggtt acggccagga
     2521 cagtcgtttg ccgtctgaat ttgacctgag cgcattttta cgcgccggag aaaaccgcct
     2581 cgcggtgatg gtgctgcgtt ggagtgacgg cagttatctg gaagatcagg atatgtggcg
     2641 gatgagcggc attttccgtg acgtctcgtt gctgcataaa ccgactacac aaatcagcga
     2701 tttccatgtt gccactcgct ttaatgatga tttcagccgc gctgtactgg aggctgaagt
     2761 tcagatgtgc ggcgagttgc gtgactacct acgggtaaca gtttctttat ggcagggtga
     2821 aacgcaggtc gccagcggca ccgcgccttt cggcggtgaa attatcgatg agcgtggtgg
     2881 ttatgccgat cgcgtcacac tacgtctgaa cgtcgaaaac ccgaaactgt ggagcgccga
     2941 aatcccgaat ctctatcgtg cggtggttga actgcacacc gccgacggca cgctgattga
     3001 agcagaagcc tgcgatgtcg gtttccgcga ggtgcggatt gaaaatggtc tgctgctgct
     3061 gaacggcaag ccgttgctga ttcgaggcgt taaccgtcac gagcatcatc ctctgcatgg
     3121 tcaggtcatg gatgagcaga cgatggtgca ggatatcctg ctgatgaagc agaacaactt
     3181 taacgccgtg cgctgttcgc attatccgaa ccatccgctg tggtacacgc tgtgcgaccg
     3241 ctacggcctg tatgtggtgg atgaagccaa tattgaaacc cacggcatgg tgccaatgaa
     3301 tcgtctgacc gatgatccgc gctggctacc ggcgatgagc gaacgcgtaa cgcgaatggt
     3361 gcagcgcgat cgtaatcacc cgagtgtgat catctggtcg ctggggaatg aatcaggcca
     3421 cggcgctaat cacgacgcgc tgtatcgctg gatcaaatct gtcgatcctt cccgcccggt
     3481 gcagtatgaa ggcggcggag ccgacaccac ggccaccgat attatttgcc cgatgtacgc
     3541 gcgcgtggat gaagaccagc ccttcccggc tgtgccgaaa tggtccatca aaaaatggct
     3601 ttcgctacct ggagagacgc gcccgctgat cctttgcgaa tacgcccacg cgatgggtaa
     3661 cagtcttggc ggtttcgcta aatactggca ggcgtttcgt cagtatcccc gtttacaggg
     3721 cggcttcgtc tgggactggg tggatcagtc gctgattaaa tatgatgaaa acggcaaccc
     3781 gtggtcggct tacggcggtg attttggcga tacgccgaac gatcgccagt tctgtatgaa
     3841 cggtctggtc tttgccgacc gcacgccgca tccagcgctg acggaagcaa aacaccagca
     3901 gcagtttttc cagttccgtt tatccgggca aaccatcgaa gtgaccagcg aatacctgtt
     3961 ccgtcatagc gataacgagc tcctgcactg gatggtggcg ctggatggta agccgctggc
     4021 aagcggtgaa gtgcctctgg atgtcgctcc acaaggtaaa cagttgattg aactgcctga
     4081 actaccgcag ccggagagcg ccgggcaact ctggctcaca gtacgcgtag tgcaaccgaa
     4141 cgcgaccgca tggtcagaag ccgggcacat cagcgcctgg cagcagtggc gtctggcgga
     4201 aaacctcagt gtgacgctcc ccgccgcgtc ccacgccatc ccgcatctga ccaccagcga
     4261 aatggatttt tgcatcgagc tgggtaataa gcgttggcaa tttaaccgcc agtcaggctt
     4321 tctttcacag atgtggattg gcgataaaaa acaactgctg acgccgctgc gcgatcagtt
     4381 cacccgtgca ccgctggata acgacattgg cgtaagtgaa gcgacccgca ttgaccctaa
     4441 cgcctgggtc gaacgctgga aggcggcggg ccattaccag gccgaagcag cgttgttgca
     4501 gtgcacggca gatacacttg ctgatgcggt gctgattacg accgctcacg cgtggcagca
     4561 tcaggggaaa accttattta tcagccggaa aacctaccgg attgatggta gtggtcaaat
     4621 ggcgattacc gttgatgttg aagtggcgag cgatacaccg catccggcgc ggattggcct
     4681 gaactgccag ctggcgcagg tagcagagcg ggtaaactgg ctcggattag ggccgcaaga
     4741 aaactatccc gaccgcctta ctgccgcctg ttttgaccgc tgggatctgc cattgtcaga
     4801 catgtatacc ccgtacgtct tcccgagcga aaacggtctg cgctgcggga cgcgcgaatt
     4861 gaattatggc ccacaccagt ggcgcggcga cttccagttc aacatcagcc gctacagtca
     4921 acagcaactg atggaaacca gccatcgcca tctgctgcac gcggaagaag gcacatggct
     4981 gaatatcgac ggtttccata tggggattgg tggcgacgac tcctggagcc cgtcagtatc
     5041 ggcggatttc cagctgagcg ccggtcgcta ccattaccag ttggtctggt gtcaaaaata
     5101 ataataaccg ggcaggccat gtctgcccgt atttcgcgta aggaaatcca ttatgtacta
     5161 tttccttgca ggccaaccag ataagtgaaa tctagttcca aactattttg tcatttttaa
     5221 ttttcgtatt agcttacgac gctacaccca gttcccatct attttgtcac tcttccctaa
     5281 ataatcctta aaaactccat ttccacccct cccagttccc aactattttg tccgcccaca
     5341 gcggggcatt tttcttcctg ttatgtttgg gcgctgcatt aatgaatcgg ccaacgcgcg
     5401 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     5461 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     5521 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5581 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5641 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5701 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5761 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     5821 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     5881 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     5941 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6001 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     6061 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6121 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     6181 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     6241 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6301 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     6361 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     6421 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     6481 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     6541 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     6601 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     6661 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     6721 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     6781 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     6841 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     6901 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     6961 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     7021 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     7081 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     7141 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     7201 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     7261 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     7321 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     7381 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtc
//
