LOCUS       sequence-151362-        4779 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template plasmid which encodes kanamycin resistance gene for 
            positive selection and toxin gene (relE) under the control of 
            arabinose induceable promoter (ParaB) for negative selection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4779)
  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 4779)
  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-237
FEATURES             Location/Qualifiers
     source          1..4779
                     /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             459..1253
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(513..532)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1123..1142
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      1530..1616
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1708..1735
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1982..2266
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     2189..2208
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     terminator      2657..2903
                     /label=lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
     protein_bind    complement(2956..2989)
                     /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      3038..3394
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(join(3491..4282,4283..4351))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4114..4133
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4352..4456)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      4547..4633
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4725..4752
                     /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 ggaacttcaa gatcccctca cgctgccgca
      121 agcactcagg gcgcaagggc tgctaaagga agcggaacac gtagaaagcc agtccgcaga
      181 aacggtgctg accccggatg aatgtcagct actgggctat ctggacaagg gaaaacgcaa
      241 gcgcaaagag aaagcaggta gcttgcagtg ggcttacatg gcgatagcta gactgggcgg
      301 ttttatggac agcaagcgaa ccggaattgc cagctggggc gccctctggt aaggttggga
      361 agccctgcaa agtaaactgg atggctttct tgccgccaag gatctgatgg cgcaggggat
      421 caagatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
      481 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
      541 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
      601 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
      661 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
      721 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
      781 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
      841 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
      901 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
      961 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     1021 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     1081 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     1141 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     1201 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     1261 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     1321 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     1381 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccagct tcaaaagcga
     1441 tcctccgcgg cccaaagtgc cacctgcatc gatggccccc gatggtagtg tggggtctcc
     1501 ccatgcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact
     1561 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc
     1621 cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc
     1681 cataaactgc caggcatcaa attaagcaga aggccatcct gacggatggc ctttttgcgt
     1741 ggccagtgcc aagcttgcat gcctgcaggt cgacggatcc ccgggtaccg agctctcgag
     1801 cgaatggtga gattgagaat ataacctttc attcccagcg gtcggtcgat aaaaaaatcg
     1861 agataaccgt tggcctcaat cggcgttaaa cccgccacca gatgggcatt aaacgagtat
     1921 cccggcagca ggggatcatt ttgcgcttca gccatacttt tcatactccc accattcaga
     1981 gaagaaacca attgtccata ttgcatcaga cattgccgtc actgcgtctt ttactggctc
     2041 ttctcgctaa ccaaaccggt aaccccgctt attaaaagca ttctgtaaca aagcgggacc
     2101 aaagccatga caaaaacgcg taacaaaagt gtctataatc acggcagaaa agtccacatt
     2161 gattatttgc acggcgtcac actttgctat gccatagcat ttttatccat aagattagcg
     2221 gatcctacct gacgcttttt atcgcaactc tctactgttt ctccatacct gcaggtaccg
     2281 gtccggaatt cttgacggag taaagccata tggcgtattt tctggatttt gacgagcggg
     2341 cactaaagga atggcgaaag ctgggctcga cggtacgtga acagttgaaa aagaagctgg
     2401 ttgaagtact tgagtcaccc cggattgaag caaacaagct ccgtggtatg cctgattgtt
     2461 acaagattaa gctccggtct tcaggctatc gccttgtata ccaggttata gacgagaaag
     2521 ttgtcgtttt cgtgatttct gttgggaaaa gagaacgctc ggaagtatat agcgaggcgg
     2581 tcaaacgcat tctctgagga tccgcgaatt ctcatgtttg acagcttatc actgatcagt
     2641 gaattaatgg cgatgacgca tcctcacgat aatatccggg taggcgcaat cactttcgtc
     2701 tctactccgt tacaaagcga ggctgggtat ttcccggcct ttctgttatc cgaaatccac
     2761 tgaaagcaca gcggctggct gaggagataa ataataaacg aggggctgta tgcacaaagc
     2821 atcttctgtt gagttaagaa cgagtatcga gatggcacat agccttgctc aaattggaat
     2881 caggtttgtg ccaataccag tagaaacaga cgaagaagct agctttgcac tggatgggga
     2941 cgtcaggatc gctctgaagt tcctatactt tctagagaat aggaacttcg gaataggaac
     3001 taaggaggat attcatatgg accatggcta attcccatgt cagccgttaa gtgttcctgt
     3061 gtcactcaaa attgctttga gaggctctaa gggcttctca gtgcgttaca tccctggctt
     3121 gttgtccaca accgttaaac cttaaaagct ttaaaagcct tatatattct tttttttctt
     3181 ataaaactta aaaccttaga ggctatttaa gttgctgatt tatattaatt ttattgttca
     3241 aacatgagag cttagtacgt gaaacatgag agcttagtac gttagccatg agagcttagt
     3301 acgttagcca tgagggttta gttcgttaaa catgagagct tagtacgtta aacatgagag
     3361 cttagtacgt gaaacatgag agcttagtac gtactatcaa caggttgaac tgcggatctt
     3421 gcggccgcaa aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3481 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     3541 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     3601 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     3661 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     3721 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     3781 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     3841 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     3901 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     3961 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     4021 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     4081 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     4141 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     4201 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     4261 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     4321 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     4381 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     4441 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgcatcgatg gccccccgat
     4501 ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa
     4561 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct
     4621 gagtaggaca aatccgccgg gagcggattt gaacgttgcg aagcaacggc ccggagggtg
     4681 gcgggcagga cgcccgccat aaactgccag gcatcaaatt aagcagaagg ccatcctgac
     4741 ggatggcctt tttgcgtggc cagtgccaag cttgcatgc
//
