LOCUS       sequence-153071-        3882 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template plasmid which encodes chloramphenicol resistance gene for 
            positive selection and a toxin gene (relE) under the control of 
            rhamnose induceable promoter (PrhaB) for negative selection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3882)
  AUTHORS   Khetrapal V, Mehershahi K, Rafee S, Chen S, Lim CL, Chen SL
  TITLE     A set of powerful negative selection systems for unmodified 
            Enterobacteriaceae.
  JOURNAL   Nucleic Acids Res. 2015 Jul 27;43(13):e83. doi: 10.1093/nar/gkv248. 
            Epub 2015 Mar 23.
  PUBMED    25800749
REFERENCE   2  (bases 1 to 3882)
  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: "Nucleic 
            Acids Res."; date: "2015-07-27"; volume: "43(13):e83. doi"; pages: "
            10.1093/nar/gkv248. Epub 2015 Mar 23"
COMMENT     Sequence Label: pSLC-242
FEATURES             Location/Qualifiers
     source          1..3882
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(51..98)
                     /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(115..774)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     689..708
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(775..877)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      1082..1168
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1260..1287
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     regulatory      1341..1350
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        1490..1608
                     /label=rhaB promoter
                     /note="promoter of the E. coli rhaBAD operon, conferring 
                     tight induction with L-rhamnose and repression with 
                     D-glucose in the presence of RhaR and RhaS (Giacalone et 
                     al., 2006)"
     terminator      1979..2225
                     /label=lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
     protein_bind    complement(2278..2311)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     rep_origin      2360..2716
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     promoter        complement(3456..3560)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      3650..3736
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3828..3855
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 agattgcagc attacacgtc ttgagcgatt gtgtaggctg gagctgcttc gaagttccta
       61 tactttctag agaataggaa cttcggaata ggaacttcat ttaaatggcg cgccttacgc
      121 cccgccctgc cactcatcgc agtactgttg taattcatta agcattctgc cgacatggaa
      181 gccatcacaa acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg
      241 cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt
      301 taaatcaaaa ctggtgaaac tcacccaggg attggctgag acgaaaaaca tattctcaat
      361 aaacccttta gggaaatagg ccaggttttc accgtaacac gccacatctt gcgaatatat
      421 gtgtagaaac tgccggaaat cgtcgtggta ttcactccag agcgatgaaa acgtttcagt
      481 ttgctcatgg aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc
      541 tttcattgcc atacgtaatt ccggatgagc attcatcagg cgggcaagaa tgtgaataaa
      601 ggccggataa aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag
      661 ctgaacggtc tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt
      721 acgatgccat tgggatatat caacggtggt atatccagtg atttttttct ccattttagc
      781 ttccttagct cctgaaaatc tcgacaactc aaaaaatacg cccggtagtg atcttatttc
      841 attatggtga aagttggaac ctcttacgtg ccgatcaacg tctcattttc gccaaaagtt
      901 ggcccagggc ttcccggtat caacagggac accaggattt atttattctg cgaagtgatc
      961 ttccgtcaca ggtaggcgcg ccgaagttcc tatactccgc ggcccaaagt gccacctgca
     1021 tcgatggccc ccgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca
     1081 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
     1141 ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca
     1201 acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc aaattaagca
     1261 gaaggccatc ctgacggatg gcctttttgc gtggccagtg ccaagcttgc atgcctgcag
     1321 ctgtccggaa ctagtcgagg ccatggtggc ctcctgatgt cgtcaacacg gcgaaatagt
     1381 aatcacgagg tcaggttctt accttaaatt ttcgacggaa aaccacgtaa aaaacgtcga
     1441 tttttcaaga tacagcgtga attttcagga aatgcggtga gcatcacatc accacaattc
     1501 agcaaattgt gaacatcatc acgttcatct ttccctggtt gccaatggcc cattttcctg
     1561 tcagtaacga gaaggtcgcg aattcaggcg ctttttagac tggtcgtaat gaaattcagc
     1621 aggatcacca tatggcgtat tttctggatt ttgacgagcg ggcactaaag gaatggcgaa
     1681 agctgggctc gacggtacgt gaacagttga aaaagaagct ggttgaagta cttgagtcac
     1741 cccggattga agcaaacaag ctccgtggta tgcctgattg ttacaagatt aagctccggt
     1801 cttcaggcta tcgccttgta taccaggtta tagacgagaa agttgtcgtt ttcgtgattt
     1861 ctgttgggaa aagagaacgc tcggaagtat atagcgaggc ggtcaaacgc attctctgag
     1921 gatccgcgaa ttctcatgtt tgacagctta tcactgatca gtgaattaat ggcgatgacg
     1981 catcctcacg ataatatccg ggtaggcgca atcactttcg tctctactcc gttacaaagc
     2041 gaggctgggt atttcccggc ctttctgtta tccgaaatcc actgaaagca cagcggctgg
     2101 ctgaggagat aaataataaa cgaggggctg tatgcacaaa gcatcttctg ttgagttaag
     2161 aacgagtatc gagatggcac atagccttgc tcaaattgga atcaggtttg tgccaatacc
     2221 agtagaaaca gacgaagaag ctagctttgc actggatggg gacgtcgatc ctgctctgaa
     2281 gttcctatac tttctagaga ataggaactt cggaatagga actaaggagg atattcatat
     2341 ggaccatggc taattcccat gtcagccgtt aagtgttcct gtgtcactga aaattgcttt
     2401 gagaggctct aagggcttct cagtgcgtta catccctggc ttgttgtcca caaccgttaa
     2461 accttaaaag ctttaaaagc cttatatatt cttttttttc ttataaaact taaaacctta
     2521 gaggctattt aagttgctga tttatattaa ttttattgtt caaacatgag agcttagtac
     2581 gtgaaacatg agagcttagt acgttagcca tgagagctta gtacgttagc catgagggtt
     2641 tagttcgtta aacatgagag cttagtacgt taaacatgag agcttagtac gtgaaacatg
     2701 agagcttagt acgtactatc aacaggttga actgcggatc ttgcggccgc aaaaattaaa
     2761 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     2821 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     2881 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     2941 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     3001 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     3061 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     3121 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     3181 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     3241 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     3301 tggcagcact gcataattct cttactgtca tgccatgcac ccaactgatc ttcagcatct
     3361 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     3421 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     3481 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     3541 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgcatc gatggccccc
     3601 gatggtagtg tggggtctcc ccatgcgaga gtagggaact gccaggcatc aaataaaatg
     3661 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct
     3721 cctgagtagg acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg
     3781 gtggcgggca ggacgcccgc cataaactgc caggcatcaa attaagcaga aggccatcct
     3841 gacggatggc ctttttgcgt ggccagtgcc aagcttgcat gc
//
