LOCUS       sequence-193889-        5871 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Suicide vector for gene replacement.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5871)
  AUTHORS   Quandt J, Hynes MF
  TITLE     Versatile suicide vectors which allow direct selection for gene 
            replacement in gram-negative bacteria.
  JOURNAL   Gene. 1993 May 15;127(1):15-21.
  PUBMED    8486283
REFERENCE   2  (bases 1 to 5871)
  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: "Gene."; 
            date: "1993-05-15"; volume: "127(1)"; pages: "15-21"
COMMENT     Sequence Label: pJQ200mp18
FEATURES             Location/Qualifiers
     source          1..5871
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(491..496,497..513,514..519))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    1347..1368
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1383..1388,1389..1406,1407..1413)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1421..1437
                     /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)."
     primer_bind     1426..1448
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1445..1461
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1445..1461
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1462..1692
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNRLA
                     AHPPFASWRNSEEARTDRPSQQLRSLNGEWR"
     misc_feature    1470..1526
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1530..1547)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1530..1546)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1539..1561)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2133..2150)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2267..2812)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(2347..2366)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     oriT            3195..3304
                     /note="incP origin of transfer"
     promoter        3680..4125
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             join(4126..4212,4213..5547)
                     /codon_start=1
                     /gene="Bacillus subtilis sacB"
                     /product="secreted levansucrase that renders bacterial 
                     growth sensitive to sucrose"
                     /label=SacB
                     /note="negative selection marker"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     CDS             complement(join(5640..5871,1..302))
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 tgctcaaact tgggcagaac gtaagccgcg agagcgccaa caaccgcttc ttggtcgaag
       61 gcagcaagcg cgatgaatgt cttactacgg agcaagttcc cgaggtaatc ggagtccggc
      121 tgatgttggg agtaggtggc tacgtctccg aactcacgac cgaaaagatc aagagcagcc
      181 cgcatggatt tgacttggtc agggccgagc ctacatgtgc gaatgatgcc catacttgag
      241 ccacctaact ttgttttagg gcgactgccc tgctgcgtaa catcgttgct gctgcgtaac
      301 atcgttgctg ctccataaca tcaaacatcg acccacggcg taacgcgctt gctgcttgga
      361 tgcccgaggc atagactgta caaaaaaaca gtcataacaa gccatgaaaa ccgccactgc
      421 gccgttacca ccgctgcgtt cggtcaaggt tctggaccag ttgcgtgagc gcatacgcta
      481 cttgcattac agtttacgaa ccgaacaggc ttatgtcaac tgggttcgtg ccttcatccg
      541 tttccacggt gtgcgtcacc cggcaacctt gggcagcagc gaagtcgagg catttctgtc
      601 ctggctggcg aacgagcgca aggtttcggt ctccacgcat cgtcaggcat tggcggcctt
      661 gctgttcttc tacggcaagg tgctgtgcac ggatctgccc tggcttcagg agatcggaag
      721 acctcggccg tcgcggcgct tgccggtggt gctgaccccg gatgaagtgg ttcgcatcct
      781 cggttttctg gaaggcgagc atcgtttgtt cgcccagctt ctgtatggaa cgggcatgcg
      841 gatcagtgag ggtttgcaac tgcgggtcaa ggatctggat ttcgatcacg gcacgatcat
      901 cgtgcgggag ggcaagggct ccaaggatcg ggccttgatg ttacccgaga gcttggcacc
      961 cagcctgcgc gagcagaatt aattccctgg cgtaatagcg aagaggcccg caccgatcgc
     1021 ccttcccaac agttgcgcag cctgaatggc gaatggcgat aagctagctt cacgctgccg
     1081 caagcactca gggcgcaagg gctgctaaag gaagcggaac acgtagaaag ccagtccgca
     1141 gaaacggtgc tgaccccgga tgaatgtcag ctactgggct atctggacaa gggaaaacgc
     1201 aagcgcaaag agaaagcagg tagcttgcag tgggcttaca tggcgatagc tagactgggc
     1261 ggttttatgg acagcaagcg aaccggaatt gccagctggc acgacaggtt tcccgactgg
     1321 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     1381 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     1441 cacacaggaa acagctatga catgattacg aattcgagct cggtacccgg ggatcctcta
     1501 gagtcgacct gcaggcatgc aagcttggca ctggccgtcg ttttacaacg tcgtgactgg
     1561 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     1621 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     1681 gaatggcgat aagctagctt cacgctgccg caagcactca gggcgcaagg gctgctaaag
     1741 gaagcggaac acgtagaaag ccagtccgca gaaacggtgc tgaccccgga tgaatgtcag
     1801 ctactgggct atctggacaa gggaaaacgc aagcgcaaag agaaagcagg tagcttgcag
     1861 tgggcttaca tggcgatagc tagactgggc ggttttatgg acagcaagcg aaccggaatt
     1921 gccaggggaa ttaattctgt ccctcctgtt cagctactga cggggtggtg cgtaacggca
     1981 aaagcaccgc cggacatcag cgctagcgga gtgtatactg gcttactatg ttggcactga
     2041 tgagggtgtc agtgaagtgc ttcatgtggc aggagaaaaa aggctgcacc ggtgcgtcag
     2101 cagaatatgt gatacaggat atattccgct tcctcgctca ctgactcgct acgctcggtc
     2161 gttcgactgc ggcgagcgga aatggcttac gaacggggcg gagatttcct ggaagatgcc
     2221 aggaagatac ttaacaggga agtgagaggg ccgcggcaaa gccgtttttc cataggctcc
     2281 gcccccctga caagcatcac gaaatctgac gctcaaatca gtggtggcga aacccgacag
     2341 gactataaag ataccaggcg tttccccctg gcggctccct cgtgcgctct cctgttcctg
     2401 cctttcggtt taccggtgtc attccgctgt tatggccgcg tttgtctcat tccacgcctg
     2461 acactcagtt ccgggtaggc agttcgctcc aagctggact gtatgcacga accccccgtt
     2521 cagtccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggaaagacat
     2581 gcaaaagcac cactggcagc agccactggt aattgattta gaggagttag tcttgaagtc
     2641 atgcgccggt taaggctaaa ctgaaaggac aagttttggt gactgcgctc ctccaagcca
     2701 gttacctcgg ttcaaagagt tggtagctca gagaaccttc gaaaaaccgc cctgcaaggc
     2761 ggttttttcg ttttcagagc aagagattac gcgcagacca aaacgatctc aagaagatca
     2821 tcttattaat cagataaaat atttctagct agagtcgatc ttcgccagca gggcgaggat
     2881 cgtggcatca ccgaaccgcg ccgtgcgcgg gtcgtcggtg agccagagtt tcagcaggcc
     2941 gcccaggcgg cccaggtcgc cattgatgcg ggccagctcg cggacgtgct catagtccac
     3001 gacgcccgtg attttgtagc cctggccgac ggccagcagg taggccgaca ggctcatgcc
     3061 ggccgccgcc gccttttcct caatcgctct tcgttcgtct ggaaggcagt acaccttgat
     3121 aggtgggctg cccttcctgg ttggcttggt ttcatcagcc atccgcttgc cctcatctgt
     3181 tacgccggcg gtagccggcc agcctcgcag agcaggattc ccgttgagca ccgccaggtg
     3241 cgaataaggg acagtgaaga aggaacaccc gctcgcgggt gggcctactt cacctatcct
     3301 gcccggctga cgccgttgga tacaccaagg aaagtctaca cgaacccttt ggcaaaatcc
     3361 tgtatatcgt gcgaaaaagg atggatatac cgaaaaaatc gctataatga ccccgaagca
     3421 gggttatgca gcggaaaagc gctgcttccc tgctgttttg tggaatatct accgactgga
     3481 aacaggcaaa tgcaggaaat tactgaactg aggggacagg cgagagacga tgccaaagag
     3541 ctacaccgac gagctggccg agtgggttga atcccgcgcg gccaagaagc gccggcgtga
     3601 tgaggctgcg gttgcgttcc tggcggtgag ggcggatgtc gatcgactct agctagagga
     3661 tcgatccttt ttaacccatc acatatacct gccgttcact attatttagt gaaatgagat
     3721 attatgatat tttctgaatt gtgattaaaa aggcaacttt atgcccatgc aacagaaact
     3781 ataaaaaata cagagaatga aaagaaacag atagattttt tagttcttta ggcccgtagt
     3841 ctgcaaatcc ttttatgatt ttctatcaaa caaaagagga aaatagacca gttgcaatcc
     3901 aaacgagagt ctaatagaat gaggtcgaaa agtaaatcgc gcgggtttgt tactgataaa
     3961 gcaggcaaga cctaaaatgt gtaaagggca aagtgtatac tttggcgtca ccccttacat
     4021 attttaggtc tttttttatt gtgcgtaact aacttgccat cttcaaacag gagggctgga
     4081 agaagcagac cgctaacaca gtacataaaa aaggagacat gaacgatgaa catcaaaaag
     4141 tttgcaaaac aagcaacagt attaaccttt actaccgcac tgctggcagg aggcgcaact
     4201 caagcgtttg cgaaagaaac gaaccaaaag ccatataagg aaacatacgg catttcccat
     4261 attacacgcc atgatatgct gcaaatccct gaacagcaaa aaaatgaaaa atatcaagtt
     4321 cctgaattcg attcgtccac aattaaaaat atctcttctg caaaaggcct ggacgtttgg
     4381 gacagctggc cattacaaaa cgctgacggc actgtcgcaa actatcacgg ctaccacatc
     4441 gtctttgcat tagccggaga tcctaaaaat gcggatgaca catcgattta catgttctat
     4501 caaaaagtcg gcgaaacttc tattgacagc tggaaaaacg ctggccgcgt ctttaaagac
     4561 agcgacaaat tcgatgcaaa tgattctatc ctaaaagacc aaacacaaga atggtcaggt
     4621 tcagccacat ttacatctga cggaaaaatc cgtttattct acactgattt ctccggtaaa
     4681 cattacggca aacaaacact gacaactgca caagttaacg tatcagcatc agacagctct
     4741 ttgaacatca acggtgtaga ggattataaa tcaatctttg acggtgacgg aaaaacgtat
     4801 caaaatgtac agcagttcat cgatgaaggc aactacagct caggcgacaa ccatacgctg
     4861 agagatcctc actacgtaga agataaaggc cacaaatact tagtatttga agcaaacact
     4921 ggaactgaag atggctacca aggcgaagaa tctttattta acaaagcata ctatggcaaa
     4981 agcacatcat tcttccgtca agaaagtcaa aaacttctgc aaagcgataa aaaacgcacg
     5041 gctgagttag caaacggcgc tctcggtatg attgagctaa acgatgatta cacactgaaa
     5101 aaagtgatga aaccgctgat tgcatctaac acagtaacag atgaaattga acgcgcgaac
     5161 gtctttaaaa tgaacggcaa atggtacctg ttcactgact cccgcggatc aaaaatgacg
     5221 attgacggca ttacgtctaa cgatatttac atgcttggtt atgtttctaa ttctttaact
     5281 ggcccataca agccgctgaa caaaactggc cttgtgttaa aaatggatct tgatcctaac
     5341 gatgtaacct ttacttactc acacttcgct gtacctcaag cgaaaggaaa caatgtcgtg
     5401 attacaagct atatgacaaa cagaggattc tacgcagaca aacaatcaac gtttgcgcca
     5461 agcttcctgc tgaacatcaa aggcaagaaa acatctgttg tcaaagacag catccttgaa
     5521 caaggacaat taacagttaa caaataaaaa cgcaaaagaa aatgccgatg gccgcggcgt
     5581 tgtgacaatt taccgaacaa ctccgcggcc gggaagccga tctcggcttg aacgaattgt
     5641 taggtggcgg tacttgggtc gatatcaaag tgcatcactt cttcccgtat gcccaacttt
     5701 gtatagagag ccactgcggg atcgtcaccg taatctgctt gcacgtagat cacataagca
     5761 ccaagcgcgt tggcctcatg cttgaggaga ttgatgagcg cggtggcaat gccctgcctc
     5821 cggtgctcgc cggagactgc gagatcatag atatagatct cactacgcgg c
//
