LOCUS       sequence-51229-e        4922 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4922)
  AUTHORS   Gueguen E, Cascales E
  TITLE     Promoter swapping unveils the role of the Citrobacter rodentium CTS1
            type VI secretion system in interbacterial competition.
  JOURNAL   Appl Environ Microbiol. 2013 Jan;79(1):32-8. doi: 
            10.1128/AEM.02504-12. Epub 2012 Oct 12.
  PUBMED    23064344
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"; doi: 
            "10.1128/AEM.02504-12"; journalName: "Appl Environ Microbiol"; date:
            "2013-01"; volume: "79"; issue: "1"; pages: "32-8"
COMMENT     Sequence Label: pRL128
FEATURES             Location/Qualifiers
     source          1..4922
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(40..62)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    51..67
                     /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(75..81,82..97,98..103))
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    complement(341..388)
                     /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(401..1195)
                     /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(512..531)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1122..1141
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     protein_bind    complement(1563..1596)
                     /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)"
     CDS             complement(1621..2499)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        2526..2810
                     /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     2733..2752
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     terminator      2874..3120
                     /label=lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
     rep_origin      3181..3537
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(join(3634..4425,4426..4494))
                     /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     4257..4276
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4495..4599)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      4690..4776
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4868..4895
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 agattgcagc attacacgtc ttgagcgatt tggctgtttc ctgtgtgaaa ttgttatccg
       61 ctcacaattc cacacattat acgagccgat gattaattgt caacagctca tttcagaata
      121 tttgccagaa ccgttatgat gtcggcgcaa aaaacattat ccagaacggg agtgcgcctt
      181 gagcgacacg aattatgcag tgatttacga cctgcacagc cataccacag cttccgatgg
      241 ctgcctgacg ccagaagcat tggtgcaccg tgcagtcgat gataagctgt caaacatgag
      301 aattgtgcct aatgagtgag gtgtaggctg gagctgcttc gaagttccta tactttctag
      361 agaataggaa cttcggaata ggaacttcaa gatcccctta ttagaagaac tcgtcaagaa
      421 ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc
      481 ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac gctatgtcct
      541 gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag cggccatttt
      601 ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc tcgccgtcgg
      661 gcatgcgcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga tgctcttcgt
      721 ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc tcgatgcgat
      781 gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc cgccgcattg
      841 catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg agatcctgcc
      901 ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg tcgagcacag
      961 ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg tcctgcagtt
     1021 cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca
     1081 gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca tagccgaata
     1141 gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa
     1201 acgatcctca tcctgtctct tgatcagatc ttgatcccct gcgccatcag atccttggcg
     1261 gcaagaaagc catccagttt actttgcagg gcttcccaac cttaccagag ggcgccccag
     1321 ctggcaattc cggttcgctt gctgtccata aaaccgccca gtctagctat cgccatgtaa
     1381 gcccactgca agctacctgc tttctctttg cgcttgcgtt ttcccttgtc cagatagccc
     1441 agtagctgac attcatccgg ggtcagcacc gtttctgcgg actggctttc tacgtgttcc
     1501 gcttccttta gcagcccttg cgccctgagt gcttgcggca gcgtgagctt caaaagcgct
     1561 ctgaagttcc tatactttct agagaatagg aacttcgaac tgcaggtcga cggatcccaa
     1621 ttatgacaac ttgacggcta catcattcac tttttcttca caaccggcac ggaactcgct
     1681 cgggctggcc ccggtgcatt ttttaaatac ccgcgagaaa tagagttgat cgtcaaaacc
     1741 aacattgcga ccgacggtgg cgataggcat ccgggtggtg ctcaaaagca gcttcgcctg
     1801 gctgatacgt tggtcctcgc gccagcttaa gacgctaatc cctaactgct ggcggaaaag
     1861 atgtgacaga cgcgacggcg acaagcaaac atgctgtgcg acgctggcga tatcaaaatt
     1921 gctgtctgcc aggtgatcgc tgatgtactg acaagcctcg cgtacccgat tatccatcgg
     1981 tggatggagc gactcgttaa tcgcttccat gcgccgcagt aacaattgct caagcagatt
     2041 tatcgccagc agctccgaat agcgcccttc cccttgcccg gcgttaatga tttgcccaaa
     2101 caggtcgctg aaatgcggct ggtgcgcttc atccgggcga aagaaccccg tattggcaaa
     2161 tattgacggc cagttaagcc attcatgcca gtaggcgcgc ggacgaaagt aaacccactg
     2221 gtgataccat tcgcgagcct ccggatgacg accgtagtga tgaatctctc ctggcgggaa
     2281 cagcaaaata tcacccggtc ggcaaacaaa ttctcgtccc tgatttttca ccaccccctg
     2341 accgcgaatg gtgagattga gaatataacc tttcattccc agcggtcggt cgataaaaaa
     2401 atcgagataa ccgttggcct caatcggcgt taaacccgcc accagatggg cattaaacga
     2461 gtatcccggc agcaggggat cattttgcgc ttcagccata cttttcatac tcccgccatt
     2521 cagagaagaa accaattgtc catattgcat cagacattgc cgtcactgcg tcttttactg
     2581 gctcttctcg ctaaccaaac cggtaacccc gcttattaaa agcattctgt aacaaagcgg
     2641 gaccaaagcc atgacaaaaa cgcgtaacaa aagtgtctat aatcacggca gaaaagtcca
     2701 cattgattat ttgcacggcg tcacactttg ctatgccata gcatttttat ccataagatt
     2761 agcggatcct acctgacgct ttttatcgca actctctact gtttctccat accccccgga
     2821 attaattctc atgtttgaca gcttatcact gatcagtgaa ttaatggcga tgacgcatcc
     2881 tcacgataat atccgggtag gcgcaatcac tttcgtctct actccgttac aaagcgaggc
     2941 tgggtatttc ccggcctttc tgttatccga aatccactga aagcacagcg gctggctgag
     3001 gagataaata ataaacgagg ggctgtatgc acaaagcatc ttctgttgag ttaagaacga
     3061 gtatcgagat ggcacatagc cttgctcaaa ttggaatcag gtttgtgcca ataccagtag
     3121 aaacagacga agaagctagc tttgcactgg attgcgaggc tttgccatgg ctaattccca
     3181 tgtcagccgt taagtgttcc tgtgtcactg aaaattgctt tgagaggctc taagggcttc
     3241 tcagtgcgtt acatccctgg cttgttgtcc acaaccgtta aaccttaaaa gctttaaaag
     3301 ccttatatat tctttttttt cttataaaac ttaaaacctt agaggctatt taagttgctg
     3361 atttatatta attttattgt tcaaacatga gagcttagta cgtgaaacat gagagcttag
     3421 tacgttagcc atgagagctt agtacgttag ccatgagggt ttagttcgtt aaacatgaga
     3481 gcttagtacg ttaaacatga gagcttagta cgtgaaacat gagagcttag tacgtactat
     3541 caacaggttg aactgcggat cttgcggccg caaaaattaa aaatgaagtt ttaaatcaat
     3601 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     3661 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     3721 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     3781 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     3841 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     3901 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     3961 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     4021 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     4081 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     4141 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     4201 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     4261 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     4321 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     4381 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     4441 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     4501 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     4561 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     4621 acctgcatcg atggcccccc gatggtagtg tggggtctcc ccatgcgaga gtagggaact
     4681 gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
     4741 tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt
     4801 gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc cataaactgc caggcatcaa
     4861 attaagcaga aggccatcct gacggatggc ctttttgcgt ggccagtgcc aagcttgcat
     4921 gc
//
