LOCUS       sequence-154517-        5213 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Suicide vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5213)
  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 5213)
  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: pJQ200KS
FEATURES             Location/Qualifiers
     source          1..5213
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(43..59)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(44..60)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(89..109)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(89..107)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(128..144)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(128..144)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(141..163)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    152..168
                     /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)."
     promoter        complement(join(176..182,183..200,201..206))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    221..242
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(501..518)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(635..1179)
                     /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(715..734)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     oriT            1553..1662
                     /note="incP origin of transfer"
     promoter        2038..2483
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             join(2484..2570,2571..3905)
                     /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(3998..4531)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGSEFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(4720..4725,4726..4742,4743..4748))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 tagtggatcc cccgggctgc aggaattcga tatcaagctt atcgataccg tcgacctcga
       61 gggggggccc ggtacccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt
      121 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
      181 tacgagccgg gagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
      241 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc caggggaatt
      301 aattctgtcc ctcctgttca gctactgacg gggtggtgcg taacggcaaa agcaccgccg
      361 gacatcagcg ctagcggagt gtatactggc ttactatgtt ggcactgatg agggtgtcag
      421 tgaagtgctt catgtggcag gagaaaaaag gctgcaccgg tgcgtcagca gaatatgtga
      481 tacaggatat attccgcttc ctcgctcact gactcgctac gctcggtcgt tcgactgcgg
      541 cgagcggaaa tggcttacga acggggcgga gatttcctgg aagatgccag gaagatactt
      601 aacagggaag tgagagggcc gcggcaaagc cgtttttcca taggctccgc ccccctgaca
      661 agcatcacga aatctgacgc tcaaatcagt ggtggcgaaa cccgacagga ctataaagat
      721 accaggcgtt tccccctggc ggctccctcg tgcgctctcc tgttcctgcc tttcggttta
      781 ccggtgtcat tccgctgtta tggccgcgtt tgtctcattc cacgcctgac actcagttcc
      841 gggtaggcag ttcgctccaa gctggactgt atgcacgaac cccccgttca gttcgaccgc
      901 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg aaagacatgc aaaagcacca
      961 ctggcagcag ccactggtaa ttgatttaga ggagttagtc ttgaagtcat gcgccggtta
     1021 aggctaaact gaaaggacaa gttttggtga ctgcgctcct ccaagccagt tacctcggtt
     1081 caaagagttg gtagctcaga gaaccttcga aaaccgccct gcaaggcggt tttttcgttt
     1141 tcagagcaag agattacgcg cagaccaaaa cgatctcaag aagatcatct tattaatcag
     1201 ataaaatatt tctagctaga gtcgatcttc gccagcaggg caggatcgtg gcatcaccga
     1261 accgcgccgt gccgctcgtc ggtgagccag agtttcagca ggccgcccag ggggaggtcg
     1321 ccattgatgc gggccagctc gcggacgtgc tcatagtcca cgacgcccgt gattttgtag
     1381 ccctggccga cgcgacaggt aggccgacag ctcatgccgg ccgccgccgc cttttcctca
     1441 atcgctcttc gttcgtctgg aaggcagtac accttgatag gtgggctgcc cttcctggtt
     1501 ggcttggttt catcagccat ccgcttgccc tcatctgtta cgccggcggt agccggccag
     1561 cctcgcagag caggattccc gttgagcacc gccaggtgcg aataagggac agtgaagaag
     1621 gaacacccgc tcgcgggtgg gcctacttca cctatcctgc ccggctgacg ccgttggata
     1681 caccaaggaa agtctacgcg aaccctttgg caaaatcctg tatatcgtgc gaaaaaggat
     1741 ggatataccg aaaaaatcgc tataatgacc ccgaagcagg gttatgcagc ggaaaagcgc
     1801 tgcttccctg ctgttttgtg gaatatctac cgactggaaa caggcaaatg caggaaatta
     1861 ctgaactgag gggacaggcg agagacgatg ccaaagagct acaccgacga gctggccgag
     1921 tgggttgaat cccgcgcggc caagaagcgc cggcgtgatg aggctgcggt tgcgttcctg
     1981 gcggtgaggg cggatgtcga tcgactctag ctagaggatc gatccttttt aacccatcac
     2041 atatacctgc cgttcactat tatttagtga aatgagatat tatgatattt tctgaattgt
     2101 gattaaaaag gcaactttat gcccatgcaa cagaaactat aaaaaataca gagaatgaaa
     2161 agaaacagat agatttttta gttctttagg cccgtagtct gcaaatcctt ttatgatttt
     2221 ctatcaaaca aaagaggaaa atagaccagt tgcaatccaa acgagagtct aatagaatga
     2281 ggtcgaaaag taaatcgcgc gggtttgtta ctgataaagc aggcaagacc taaaatgtgt
     2341 aaagggcaaa gtgtatactt tggcgtcacc ccttacatat tttaggtctt tttttattgt
     2401 gcgtaactaa cttgccatct tcaaacagga gggctggaag aagcagaccg ctaacacagt
     2461 acataaaaaa ggagacatga acgatgaaca tcaaaaagtt tgcaaaacaa gcaacagtat
     2521 taacctttac taccgcactg ctggcaggag gcgcaactca agcgtttgcg aaagaaacga
     2581 accaaaagcc atataaggaa acatacggca tttcccatat tacacgccat gatatgctgc
     2641 aaatccctga acagcaaaaa aatgaaaaat atcaagttcc tgaattcgat tcgtccacaa
     2701 ttaaaaatat ctcttctgca aaaggcctgg acgtttggga cagctggcca ttacaaaacg
     2761 ctgacggcac tgtcgcaaac tatcacggct accacatcgt ctttgcatta gccggagatc
     2821 ctaaaaatgc ggatgacaca tcgatttaca tgttctatca aaaagtcggc gaaacttcta
     2881 ttgacagctg gaaaaacgct ggccgcgtct ttaaagacag cgacaaattc gatgcaaatg
     2941 attctatcct aaaagaccaa acacaagaat ggtcaggttc agccacattt acatctgacg
     3001 gaaaaatccg tttattctac actgatttct ccggtaaaca ttacggcaaa caaacactga
     3061 caactgcaca agttaacgta tcagcatcag acagctcttt gaacatcaac ggtgtagagg
     3121 attataaatc aatctttgac ggtgacggaa aaacgtatca aaatgtacag cagttcatcg
     3181 atgaaggcaa ctacagctca ggcgacaacc atacgctgag agatcctcac tacgtagaag
     3241 ataaaggcca caaatactta gtatttgaag caaacactgg aactgaagat ggctaccaag
     3301 gcgaagaatc tttatttaac aaagcatact atggcaaaag cacatcattc ttccgtcaag
     3361 aaagtcaaaa acttctgcaa agcgataaaa aacgcacggc tgagttagca aacggcgctc
     3421 tcggtatgat tgagctaaac gatgattaca cactgaaaaa agtgatgaaa ccgctgattg
     3481 catctaacac agtaacagat gaaattgaac gcgcgaacgt ctttaaaatg aacggcaaat
     3541 ggtacctgtt cactgactcc cgcggatcaa aaatgacgat tgacggcatt acgtctaacg
     3601 atatttacat gcttggttat gtttctaatt ctttaactgg cccatacaag ccgctgaaca
     3661 aaactggcct tgtgttaaaa atggatcttg atcctaacga tgtaaccttt acttactcac
     3721 acttcgctgt acctcaagcg aaaggaaaca atgtcgtgat tacaagctat atgacaaaca
     3781 gaggattcta cgcagacaaa caatcaacgt ttgcgccaag cttcctgctg aacatcaaag
     3841 gcaagaaaac atctgttgtc aaagacagca tccttgaaca aggacaatta acagttaaca
     3901 aataaaaacg caaaagaaaa tgccgatggc cgcggcgttg tgacaattta ccgaacaact
     3961 ccgcggccgg gaagccgatc tcggcttgaa cgaattgtta ggtggcggta cttgggtcga
     4021 tatcaaagtg catcacttct tcccgtatgc ccaactttgt atagagagcc actgcgggat
     4081 cgtcaccgta atctgcttgc acgtagatca cataagcacc aagcgcgttg gcctcatgct
     4141 tgaggagatt gatgagcgcg gtggcaatgc cctgcctccg gtgctcgccg gagactgcga
     4201 gatcatagat atagatctca ctacgcggct gctcaaacct gggcagaacg taagccgcga
     4261 gagcgccaac aaccgcttct tggtcgaagg cagcaagcgc gatgaatgtc ttactacgga
     4321 gcaagttccc gaggtaatcg gagtccggct gatgttggga gtaggtggct acgtctccga
     4381 actcactacc gaaaagatca agagcagccc gcatggattt gacttggtca gggccgagcc
     4441 tacatgtgcg aatgatgccc atacttgagc cacctaactt tgttttaggg cgactgccct
     4501 gctgcgtaac atcgttgctg ctgcgtaaca tcgttgctgc tccataacat gaaacatcga
     4561 cccacggcgt aacgcgcttg ctgcttggat gcccgaggca tagactgtac aaaaaaacag
     4621 tcataacaag ccatgaaaac cgccactgcg ccgttaccac cgctgcgttc ggtcaaggtt
     4681 ctggaccagt tgcgtgagcg catacgctac ttgcattaca gtttacgaac cgaacaggct
     4741 tatgtcaact gggttcgtgc cttcatccgt ttccacggtg tgcgtcaccc ggcaaccttg
     4801 ggcagcagcg aagtcgaggc atttctgtcc tggctggcga acgagcgcaa ggtttcggtc
     4861 tccacgcatc gtcaggcatt ggcggccaag ctgttcttct acggcaaggt gctgtgcacg
     4921 gatctgccct ggcttcagga gatcggaaga cctcggccgt cgcggcgctt gccggtggtg
     4981 ctgaccccgg atgaagtggt tcgcatcctc ggttttctgg aaggcgagca tcgtttgttc
     5041 gcccagcttc tgtatggaac gggcatgcgg atcagtgggt ttgcaactgc gggtcaaggt
     5101 ctggatttcg atcacggcac gatcatgctg cgggagggca agggctccaa ggatcgggcc
     5161 ttgatgttac ccgagagctt ggcacccacg ctgcgcgagc agaattaatt ccc
//
