LOCUS       sequence-146447-        4922 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Base vector for construction of series of suicide plasmids for gene 
            replacement in Proteobacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4922)
  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 4922)
  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: pJQ200
FEATURES             Location/Qualifiers
     source          1..4922
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(208..225)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(342..885)
                     /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(422..441)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     oriT            1259..1368
                     /note="incP origin of transfer"
     promoter        1744..2189
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             join(2190..2276,2277..3614)
                     /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
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYRGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGISSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTS
                     VVKDSILEQGQLTVNK"
     CDS             complement(3707..4240)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGSEFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(4429..4434,4435..4451,4452..4457))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 gggaattaat tctgtccctc ctgttcagct actgacgggg tggtgcgtaa cggcaaaagc
       61 accgccggac atcagcgcta gcggagtgta tactggctta ctatgttggc actgatgagg
      121 gtgtcagtga agtgcttcat gtggcaggag aaaaaaggct gcaccggtgc gtcagcagaa
      181 tatgtgatac aggatatatt ccgcttcctc gctcactgac tcgctacgct cggtcgttcg
      241 actgcggcga gcggaaatgg cttacgaacg gggcggagat ttcctggaag atgccaggaa
      301 gatacttaac agggaagtga gagggccgcg gcaaagccgt ttttccatag gctccgcccc
      361 cctgacaagc atcacgaaat ctgacgctca aatcagtggt ggcgaaaccc gacaggacta
      421 taaagatacc aggcgtttcc cctggcggct ccctcgtgcg ctctcctgtt cctgcctttc
      481 ggtttaccgg tgtcattccg ctgttatggc cgcgtttgtc tcattccacg cctgacactc
      541 agttccgggt aggcagttcg ctccaagctg gactgtatgc acgaaccccc cgttcagttc
      601 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggaaag acatgcaaaa
      661 gcaccactgg cagcagccac tggtaattga tttagaggag ttagtcttga agtcatgcgc
      721 cggttaaggc taaactgaaa ggacaagttt tggtgactgc gctcctccaa gccagttacc
      781 tcggttcaaa gagttggtag ctcagagaac cttcgaaaac cgccctgcaa ggcggttttt
      841 tcgttttcag agcaagagat tacgcgcaga ccaaaacgat ctcaagaaga tcatcttatt
      901 aatcagataa aatatttcta gctagagtcg atcttcgcca gcagggcagg atcgtggcat
      961 caccgaaccg cgccgtgccg ctcgtcggtg agccagagtt tcagcaggcc gcccaggggg
     1021 aggtcgccat tgatgcgggc cagctcgcgg acgtgctcat agtccacgac gcccgtgatt
     1081 ttgtagccct ggccgacgcg acaggtaggc cgacagctca tgccggccgc cgccgccttt
     1141 tcctcaatcg ctcttcgttc gtctggaagg cagtacacct tgataggtgg gctgcccttc
     1201 ctggttggct tggtttcatc agccatccgc ttgccctcat ctgttacgcc ggcggtagcc
     1261 ggccagcctc gcagagcagg attcccgttg agcaccgcca ggtgcgaata agggacagtg
     1321 aagaaggaac acccgctcgc gggtgggcct acttcaccta tcctgcccgg ctgacgccgt
     1381 tggatacacc aaggaaagtc tacgcgaacc ctttggcaaa atcctgtata tcgtgcgaaa
     1441 aaggatggat ataccgaaaa aatcgctata atgaccccga agcagggtta tgcagcggaa
     1501 aagcgctgct tccctgctgt tttgtggaat atctaccgac tggaaacagg caaatgcagg
     1561 aaattactga actgagggga caggcgagag acgatgccaa agagctacac cgacgagctg
     1621 gccgagtggg ttgaatcccg cgcggccaag aagcgccggc gtgatgaggc tgcggttgcg
     1681 ttcctggcgg tgagggcgga tgtcgatcga ctctagctag aggatcgatc ctttttaacc
     1741 catcacatat acctgccgtt cactattatt tagtgaaatg agatattatg atattttctg
     1801 aattgtgatt aaaaaggcaa ctttatgccc atgcaacaga aactataaaa aatacagaga
     1861 atgaaaagaa acagatagat tttttagttc tttaggcccg tagtctgcaa atccttttat
     1921 gattttctat caaacaaaag aggaaaatag accagttgca atccaaacga gagtctaata
     1981 gaatgaggtc gaaaagtaaa tcgcgcgggt ttgttactga taaagcaggc aagacctaaa
     2041 atgtgtaaag ggcaaagtgt atactttggc gtcacccctt acatatttta ggtctttttt
     2101 tattgtgcgt aactaacttg ccatcttcaa acaggagggc tggaagaagc agaccgctaa
     2161 cacagtacat aaaaaaggag acatgaacga tgaacatcaa aaagtttgca aaacaagcaa
     2221 cagtattaac ctttactacc gcactgctgg caggaggcgc aactcaagcg tttgcgaaag
     2281 aaacgaacca aaagccatat aaggaaacat acggcatttc ccatattaca cgccatgata
     2341 tgctgcaaat ccctgaacag caaaaaaatg aaaaatatca agttcctgaa ttcgattcgt
     2401 ccacaattaa aaatatctct tctgcaaaag gcctggacgt ttgggacagc tggccattac
     2461 aaaacgctga cggcactgtc gcaaactatc gcggctacca catcgtcttt gcattagccg
     2521 gagatcctaa aaatgcggat gacacatcga tttacatgtt ctatcaaaaa gtcggcgaaa
     2581 cttctattga cagctggaaa aacgctggcc gcgtctttaa agacagcgac aaattcgatg
     2641 caaatgattc tatcctaaaa gaccaaacac aagaatggtc aggttcagcc acatttacat
     2701 ctgacggaaa aatccgttta ttctacactg atttctccgg taaacattac ggcaaacaaa
     2761 cactgacaac tgcacaagtt aacgtatcag catcagacag ctctttgaac atcaacggtg
     2821 tagaggatta taaatcaatc tttgacggtg acggaaaaac gtatcaaaat gtacagcagt
     2881 tcatcgatga aggcaactac agctcaggcg acaaccatac gctgagagat cctcactacg
     2941 tagaagataa aggccacaaa tacttagtat ttgaagcaaa cactggaact gaagatggct
     3001 accaaggcga agaatcttta tttaacaaag catactatgg caaaagcaca tcattcttcc
     3061 gtcaagaaag tcaaaaactt ctgcaaagcg ataaaaaacg cacggctgag ttagcaaacg
     3121 gcgctctcgg tatgattgag ctaaacgatg attacacact gaaaaaagtg atgaaaccgc
     3181 tgattgcatc taacacagta acagatgaaa ttgaacgcgc gaacgtcttt aaaatgaacg
     3241 gcaaatggta cctgttcact gactcccgcg gatcaaaaat gacgattgac ggcatttcgt
     3301 ctaacgatat ttacatgctt ggttatgttt ctaattcttt aactggccca tacaagccgc
     3361 tgaacaaaac tggccttgtg ttaaaaatgg atcttgatcc taacgatgta acctttactt
     3421 actcacactt cgctgtacct caagcgaaag gaaacaatgt cgtggtgatt acaagctata
     3481 tgacaaacag aggattctac gcagacaaac aatcaacgtt tgcgccaagc ttcctgctga
     3541 acatcaaagg caagaaaaca tctgttgtca aagacagcat ccttgaacaa ggacaattaa
     3601 cagttaacaa ataaaaacgc aaaagaaaat gccgatggcc gcggcgttgt gacaatttac
     3661 cgaacaactc cgcggccggg aagccgatct cggcttgaac gaattgttag gtggcggtac
     3721 ttgggtcgat atcaaagtgc atcacttctt cccgtatgcc caactttgta tagagagcca
     3781 ctgcgggatc gtcaccgtaa tctgcttgca cgtagatcac ataagcacca agcgcgttgg
     3841 cctcatgctt gaggagattg atgagcgcgg tggcaatgcc ctgcctccgg tgctcgccgg
     3901 agactgcgag atcatagata tagatctcac tacgcggctg ctcaaacctg ggcagaacgt
     3961 aagccgcgag agcgccaaca accgcttctt ggtcgaaggc agcaagcgcg atgaatgtct
     4021 tactacggag caagttcccg aggtaatcgg agtccggctg atgttgggag taggtggcta
     4081 cgtctccgaa ctcactaccg aaaagatcaa gagcagcccg catggatttg acttggtcag
     4141 ggccgagcct acatgtgcga atgatgccca tacttgagcc acctaacttt gttttagggc
     4201 gactgccctg ctgcgtaaca tcgttgctgc tgcgtaacat cgttgctgct ccataacatg
     4261 aaacatcgac ccacggcgta acgcgcttgc tgcttggatg cccgaggcat agactgtaca
     4321 aaaaaacagt cataacaagc catgaaaacc gccactgcgc cgttaccacc gctgcgttcg
     4381 gtcaaggttc tggaccagtt gcgtgagcgc atacgctact tgcattacag tttacgaacc
     4441 gaacaggctt atgtcaactg ggttcgtgcc ttcatccgtt tccacggtgt gcgtcacccg
     4501 gcaaccttgg gcagcagcga agtcgaggca tttctgtcct ggctggcgaa cgagcgcaag
     4561 gtttcggtct ccacgcatcg tcaggcattg gcggccaagc tgttcttcta cggcaaggtg
     4621 ctgtgcacgg atctgccctg gcttcaggag atcggaagac ctcggccgtc gcggcgcttg
     4681 ccggtggtgc tgaccccgga tgaagtggtt cgcatcctcg gttttctgga aggcgagcat
     4741 cgtttgttcg cccagcttct gtatggaacg ggcatgcgga tcagtgggtt tgcaactgcg
     4801 ggtcaaggtc tggatttcga tcacggcacg atcatgctgc gggagggcaa gggctccaag
     4861 gatcgggcct tgatgttacc cgagagcttg gcacccacgc tgcgcgagca gaattaattc
     4921 cc
//
