LOCUS       sequence-154514-        5217 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 5217)
  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 5217)
  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: pJQ200SK
FEATURES             Location/Qualifiers
     source          1..5217
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(37..53)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(37..53)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(90..110)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(90..108)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(129..145)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(129..145)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(142..164)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    153..169
                     /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(177..183,184..201,202..207))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    222..243
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(502..519)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(636..1180)
                     /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(716..735)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     oriT            1554..1663
                     /note="incP origin of transfer"
     promoter        2039..2484
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             join(2485..2571,2572..3909)
                     /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(4002..4535)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGSEFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(4724..4729,4730..4746,4747..4752))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 tcgataagct tgatatcgaa ttcctgcagc ccgggggatc cactagttct agagcggccg
       61 ccaccgcggt ggagctccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg
      121 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
      181 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
      241 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccaggggaat
      301 taattctgtc cctcctgttc agctactgac ggggtggtgc gtaacggcaa aagcaccgcc
      361 ggacatcagc gctagcggag tgtatactgg cttactatgt tggcactgat gagggtgtca
      421 gtgaagtgct tcatgtggca ggagaaaaaa ggctgcaccg gtgcgtcagc agaatatgtg
      481 atacaggata tattccgctt cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg
      541 gcgagcggaa atggcttacg aacggggcgg agatttcctg gaagatgcca ggaagatact
      601 taacagggaa gtgagagggc cgcggcaaag ccgtttttcc ataggctccg cccccctgac
      661 aagcatcacg aaatctgacg ctcaaatcag tggtggcgaa acccgacagg actataaaga
      721 taccaggcgt ttccccctgg cggctccctc gtgcgctctc ctgttcctgc ctttcggttt
      781 accggtgtca ttccgctgtt atggccgcgt ttgtctcatt ccacgcctga cactcagttc
      841 cgggtaggca gttcgctcca agctggactg tatgcacgaa ccccccgttc agttcgaccg
      901 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gaaagacatg caaaagcacc
      961 actggcagca gccactggta attgatttag aggagttagt cttgaagtca tgcgccggtt
     1021 aaggctaaac tgaaaggaca agttttggtg actgcgctcc tccaagccag ttacctcggt
     1081 tcaaagagtt ggtagctcag agaaccttcg aaaaccgccc tgcaaggcgg ttttttcgtt
     1141 ttcagagcaa gagattacgc gcagaccaaa acgatctcaa gaagatcatc ttattaatca
     1201 gataaaatat ttctagctag agtcgatctt cgccagcagg gcaggatcgt ggcatcaccg
     1261 aaccgcgccg tgccgctcgt cggtgagcca gagtttcagc aggccgccca gggggaggtc
     1321 gccattgatg cgggccagct cgcggacgtg ctcatagtcc acgacgcccg tgattttgta
     1381 gccctggccg acgcgacagg taggccgaca gctcatgccg gccgccgccg ccttttcctc
     1441 aatcgctctt cgttcgtctg gaaggcagta caccttgata ggtgggctgc ccttcctggt
     1501 tggcttggtt tcatcagcca tccgcttgcc ctcatctgtt acgccggcgg tagccggcca
     1561 gcctcgcaga gcaggattcc cgttgagcac cgccaggtgc gaataaggga cagtgaagaa
     1621 ggaacacccg ctcgcgggtg ggcctacttc acctatcctg cccggctgac gccgttggat
     1681 acaccaagga aagtctacgc gaaccctttg gcaaaatcct gtatatcgtg cgaaaaagga
     1741 tggatatacc gaaaaaatcg ctataatgac cccgaagcag ggttatgcag cggaaaagcg
     1801 ctgcttccct gctgttttgt ggaatatcta ccgactggaa acaggcaaat gcaggaaatt
     1861 actgaactga ggggacaggc gagagacgat gccaaagagc tacaccgacg agctggccga
     1921 gtgggttgaa tcccgcgcgg ccaagaagcg ccggcgtgat gaggctgcgg ttgcgttcct
     1981 ggcggtgagg gcggatgtcg atcgactcta gctagaggat cgatcctttt taacccatca
     2041 catatacctg ccgttcacta ttatttagtg aaatgagata ttatgatatt ttctgaattg
     2101 tgattaaaaa ggcaacttta tgcccatgca acagaaacta taaaaaatac agagaatgaa
     2161 aagaaacaga tagatttttt agttctttag gcccgtagtc tgcaaatcct tttatgattt
     2221 tctatcaaac aaaagaggaa aatagaccag ttgcaatcca aacgagagtc taatagaatg
     2281 aggtcgaaaa gtaaatcgcg cgggtttgtt actgataaag caggcaagac ctaaaatgtg
     2341 taaagggcaa agtgtatact ttggcgtcac cccttacata ttttaggtct ttttttattg
     2401 tgcgtaacta acttgccatc ttcaaacagg agggctggaa gaagcagacc gctaacacag
     2461 tacataaaaa aggagacatg aacgatgaac atcaaaaagt ttgcaaaaca agcaacagta
     2521 ttaaccttta ctaccgcact gctggcagga ggcgcaactc aagcgtttgc gaaagaaacg
     2581 aaccaaaagc catataagga aacatacggc atttcccata ttacacgcca tgatatgctg
     2641 caaatccctg aacagcaaaa aaatgaaaaa tatcaagttc ctgaattcga ttcgtccaca
     2701 attaaaaata tctcttctgc aaaaggcctg gacgtttggg acagctggcc attacaaaac
     2761 gctgacggca ctgtcgcaaa ctatcgcggc taccacatcg tctttgcatt agccggagat
     2821 cctaaaaatg cggatgacac atcgatttac atgttctatc aaaaagtcgg cgaaacttct
     2881 attgacagct ggaaaaacgc tggccgcgtc tttaaagaca gcgacaaatt cgatgcaaat
     2941 gattctatcc taaaagacca aacacaagaa tggtcaggtt cagccacatt tacatctgac
     3001 ggaaaaatcc gtttattcta cactgatttc tccggtaaac attacggcaa acaaacactg
     3061 acaactgcac aagttaacgt atcagcatca gacagctctt tgaacatcaa cggtgtagag
     3121 gattataaat caatctttga cggtgacgga aaaacgtatc aaaatgtaca gcagttcatc
     3181 gatgaaggca actacagctc aggcgacaac catacgctga gagatcctca ctacgtagaa
     3241 gataaaggcc acaaatactt agtatttgaa gcaaacactg gaactgaaga tggctaccaa
     3301 ggcgaagaat ctttatttaa caaagcatac tatggcaaaa gcacatcatt cttccgtcaa
     3361 gaaagtcaaa aacttctgca aagcgataaa aaacgcacgg ctgagttagc aaacggcgct
     3421 ctcggtatga ttgagctaaa cgatgattac acactgaaaa aagtgatgaa accgctgatt
     3481 gcatctaaca cagtaacaga tgaaattgaa cgcgcgaacg tctttaaaat gaacggcaaa
     3541 tggtacctgt tcactgactc ccgcggatca aaaatgacga ttgacggcat ttcgtctaac
     3601 gatatttaca tgcttggtta tgtttctaat tctttaactg gcccatacaa gccgctgaac
     3661 aaaactggcc ttgtgttaaa aatggatctt gatcctaacg atgtaacctt tacttactca
     3721 cacttcgctg tacctcaagc gaaaggaaac aatgtcgtgg tgattacaag ctatatgaca
     3781 aacagaggat tctacgcaga caaacaatca acgtttgcgc caagcttcct gctgaacatc
     3841 aaaggcaaga aaacatctgt tgtcaaagac agcatccttg aacaaggaca attaacagtt
     3901 aacaaataaa aacgcaaaag aaaatgccga tggccgcggc gttgtgacaa tttaccgaac
     3961 aactccgcgg ccgggaagcc gatctcggct tgaacgaatt gttaggtggc ggtacttggg
     4021 tcgatatcaa agtgcatcac ttcttcccgt atgcccaact ttgtatagag agccactgcg
     4081 ggatcgtcac cgtaatctgc ttgcacgtag atcacataag caccaagcgc gttggcctca
     4141 tgcttgagga gattgatgag cgcggtggca atgccctgcc tccggtgctc gccggagact
     4201 gcgagatcat agatatagat ctcactacgc ggctgctcaa acctgggcag aacgtaagcc
     4261 gcgagagcgc caacaaccgc ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta
     4321 cggagcaagt tcccgaggta atcggagtcc ggctgatgtt gggagtaggt ggctacgtct
     4381 ccgaactcac taccgaaaag atcaagagca gcccgcatgg atttgacttg gtcagggccg
     4441 agcctacatg tgcgaatgat gcccatactt gagccaccta actttgtttt agggcgactg
     4501 ccctgctgcg taacatcgtt gctgctgcgt aacatcgttg ctgctccata acatgaaaca
     4561 tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga ggcatagact gtacaaaaaa
     4621 acagtcataa caagccatga aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa
     4681 ggttctggac cagttgcgtg agcgcatacg ctacttgcat tacagtttac gaaccgaaca
     4741 ggcttatgtc aactgggttc gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac
     4801 cttgggcagc agcgaagtcg aggcatttct gtcctggctg gcgaacgagc gcaaggtttc
     4861 ggtctccacg catcgtcagg cattggcggc caagctgttc ttctacggca aggtgctgtg
     4921 cacggatctg ccctggcttc aggagatcgg aagacctcgg ccgtcgcggc gcttgccggt
     4981 ggtgctgacc ccggatgaag tggttcgcat cctcggtttt ctggaaggcg agcatcgttt
     5041 gttcgcccag cttctgtatg gaacgggcat gcggatcagt gggtttgcaa ctgcgggtca
     5101 aggtctggat ttcgatcacg gcacgatcat gctgcgggag ggcaagggct ccaaggatcg
     5161 ggccttgatg ttacccgaga gcttggcacc cacgctgcgc gagcagaatt aattccc
//
