LOCUS       sequence-426110-        5915 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  biphasic target plasmid version 1.0.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5915)
  AUTHORS   Glass DS, Bren A, Vaisbourd E, Mayo A, Alon U
  TITLE     A synthetic differentiation circuit in Escherichia coli for 
            suppressing mutant takeover.
  JOURNAL   Cell. 2024 Jan 31:S0092-8674(24)00061-8. doi: 
            10.1016/j.cell.2024.01.024.
  PUBMED    38320549
REFERENCE   2  (bases 1 to 5915)
  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: "Cell. 2024 
            Jan 31:S0092-8674(24)00061-8. doi: 10.1016/j.cell.2024.01.024."
COMMENT     Sequence Label: pDSG545
FEATURES             Location/Qualifiers
     source          1..5915
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1..879)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        906..1190
                     /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     1113..1132
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             1276..1287
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1294..1530
                     /codon_start=1
                     /gene="dhfrIIb from plasmid R388"
                     /product="E. coli plasmid-associated dihydrofolate 
                     reductase"
                     /label=TpR
                     /note="confers resistance to trimethoprim"
                     /translation="MGQSSDEANAPVAGQFALPLSATFGLGDRVRKKSGAAWQGQVVGW
                     YCTKLTPEGYAVESESHPGSVQIYPVAALERVA"
     misc_feature    3253..3273
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      3274..3331
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     CDS             complement(join(3427..4218,4219..4287))
                     /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     4050..4069
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4288..4359)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4813..5358)
                     /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(4893..4912)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5826..5844
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(5848..5891)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    5894..5915
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 ttatgacaac ttgacggcta catcattcac tttttcttca caaccggcac ggaactcgct
       61 cgggctggcc ccggtgcatt ttttaaatac ccgcgagaaa tagagttgat cgtcaaaacc
      121 aacattgcga ccgacggtgg cgataggcat ccgggtggtg ctcaaaagca gcttcgcctg
      181 gctgatacgt tggtcctcgc gccagcttaa gacgctaatc cctaactgct ggcggaaaag
      241 atgtgacaga cgcgacggcg acaagcaaac atgctgtgcg acgctggcga tatcaaaatt
      301 gctgtctgcc aggtgatcgc tgatgtactg acaagcctcg cgtacccgat tatccatcgg
      361 tggatggagc gactcgttaa tcgcttccat gcgccgcagt aacaattgct caagcagatt
      421 tatcgccagc agctccgaat agcgcccttc cccttgcccg gcgttaatga tttgcccaaa
      481 caggtcgctg aaatgcggct ggtgcgcttc atccgggcga aagaaccccg tattggcaaa
      541 tattgacggc cagttaagcc attcatgcca gtaggcgcgc ggacgaaagt aaacccactg
      601 gtgataccat tcgcgagcct ccggatgacg accgtagtga tgaatctctc ctggcgggaa
      661 cagcaaaata tcacccggtc ggcaaacaaa ttctcgtccc tgatttttca ccaccccctg
      721 accgcgaatg gtgagattga gaatataacc tttcattccc agcggtcggt cgataaaaaa
      781 atcgagataa ccgttggcct caatcggcgt taaacccgcc accagatggg cattaaacga
      841 gtatcccggc agcaggggat cattttgcgc ttcagccata cttttcatac tcccgccatt
      901 cagagaagaa accaattgtc catattgcat cagacattgc cgtcactgcg tcttttactg
      961 gctcttctcg ctaaccaaac cggtaacccc gcttattaaa agcattctgt aacaaagcgg
     1021 gaccaaagcc atgacaaaaa cgcgtaacaa aagtgtctat aatcacggca gaaaagtcca
     1081 cattgattat ttgcacggcg tcacactttg ctatgccata gcatttttat ccataagatt
     1141 agcggatcct acctgacgct ttttatcgca actctctact gtttctccat acccgttttt
     1201 ttgggctagc tactagagaa cttttcggat caagctatga aggacgcaaa gagggaacta
     1261 aacactttac tagagaaaga ggagaaatac tagatgggtc aaagtagcga tgaagccaac
     1321 gctcccgttg cagggcagtt tgcgcttccc ctgagtgcca cctttggctt aggggatcgc
     1381 gtacgcaaga aatctggtgc cgcttggcag ggtcaagtcg tcggttggta ttgcacaaaa
     1441 ctcactcctg aaggctatgc ggtcgagtcc gaatcccacc caggctcagt gcaaatttat
     1501 cctgtggctg cacttgaacg tgtggcctaa taactcggta ccaaattcca gaaaagaggc
     1561 ctcccgaaag gggggccttt tttcgttttg gtccgcatcc acctgtgcgg acctcggtac
     1621 caaattccag aaaagagacg ctttcgagcg tcttttttcg ttttggtcca tcgagtggcg
     1681 cgcttagttc ctcgttttct ctcgttggaa gaagaagaaa cgagaaacta aatactagag
     1741 tcacacagga ctactagatg atgcaaaccg ttttagcgaa aatcgtcgca gacaaggcga
     1801 tttgggtaga agcccgcaaa cagcagcaac cgctggccag ttttcagaat gaggttcagc
     1861 cgagcacgcg acatttttat gatgcgctac agggtgcgcg cacggcgttt attctggagt
     1921 gcaagaaagc gtcgccgtca aaaggcgtga tccgtgatga tttcgatcca gcacgcattg
     1981 ccgccattta taaacattac gcttcggcaa tttcggtgct gactgatgag aaatattttc
     2041 aggggagctt taatttcctc cccatcgtca gccaaatcgc cccgcagccg attttatgta
     2101 aagacttcat tatcgaccct taccagatct atctggcgcg ctattaccag gccgatgcct
     2161 gcttattaat gctttcagta ctggatgacg accaatatcg ccagcttgcc gccgtcgctc
     2221 acagtctgga gatgggggtg ctgaccgaag tcagtaatga agaggaacag gagcgcgcca
     2281 ttgcattggg agcaaaggtc gttggcatca acaaccgcga tctgcgtgat ttgtcgattg
     2341 atctcaaccg tacccgcgag cttgcgccga aactggggca caacgtgacg gtaatcagcg
     2401 aatccggcat caatacttac gctcaggtgc gcgagttaag ccacttcgct aacggttttc
     2461 tgattggttc ggcgttgatg gcccatgacg atttgcacgc cgccgtgcgc cgggtgttgc
     2521 tgggtgagaa taaagtatgt ggcctgacgc gtgggcaaga tgctaaagca gcttatgacg
     2581 cgggcgcgat ttacggtggg ttgatttttg ttgcgacatc accgcgttgc gtcaacgttg
     2641 aacaggcgca ggaagtgatg gctgcggcac cgttgcagta tgttggcgtg ttccgcaatc
     2701 acgatattgc cgatgtggtg gacaaagcta aggtgttatc gctggcggca gtgcaactgc
     2761 atggtaatga agaacagctg tatatcgata cgctgcgtga agctctgcca gcacatgttg
     2821 ccatctggaa agcattaagc gtcggtgaaa ccctgcccgc ccgcgagttt cagcacgttg
     2881 ataaatatgt tttagacaac ggccagggtg gaagcgggca acgttttgac tggtcactat
     2941 taaatggtca atcgcttggc aacgttctgc tggcgggggg cttaggcgca gataactgcg
     3001 tggaagcggc acaaaccggc tgcgccggac ttgattttaa ttctgctgta gagtcgcaac
     3061 cgggcatcaa agacgcacgt cttttggcct cggttttcca gacgctgcgc gcatattaat
     3121 aactcggtac caaagacgaa caataagacg ctgaaaagcg tcttttttcg ttttggtcct
     3181 ttgaagttta ggggatagtc caattattga aggcctccct aacggggggc ctttttttgt
     3241 ttctggtctc cctactagta gcggccgctg cagtccggca aaaaagggca aggtgtcacc
     3301 accctgccct ttttctttaa aaccgaaaag attacttcgc gttatgcagg cttcctcgct
     3361 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     3421 ggtaatttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     3481 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     3541 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     3601 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     3661 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     3721 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     3781 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     3841 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     3901 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     3961 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     4021 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     4081 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     4141 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     4201 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     4261 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     4321 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagatca cgcatcttcc
     4381 cgacaacgca gaccgttccg tggcaaagca aaagttcaaa atcaccaact ggtccaccta
     4441 caacaaagct ctcatcaacc gtggctccct cactttctgg ctggatgatg gggcgattca
     4501 ggcctggtat gagtcagcaa caccttcttc acgaggcaga cctcagcgct agcggagtgt
     4561 atactggctt actatgttgg cactgatgag ggtgtcagtg aagtgcttca tgtggcagga
     4621 gaaaaaaggc tgcaccggtg cgtcagcaga atatgtgata caggatatat tccgcttcct
     4681 cgctcactga ctcgctacgc tcggtcgttc gactgcggcg agcggaaatg gcttacgaac
     4741 ggggcggaga tttcctggaa gatgccagga agatacttaa cagggaagtg agagggccgc
     4801 ggcaaagccg tttttccata ggctccgccc ccctgacaag catcacgaaa tctgacgctc
     4861 aaatcagtgg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggcgg
     4921 ctccctcgtg cgctctcctg ttcctgcctt tcggtttacc ggtgtcattc cgctgttatg
     4981 gccgcgtttg tctcattcca cgcctgacac tcagttccgg gtaggcagtt cgctccaagc
     5041 tggactgtat gcacgaaccc cccgttcagt ccgaccgctg cgccttatcc ggtaactatc
     5101 gtcttgagtc caacccggaa agacatgcaa aagcaccact ggcagcagcc actggtaatt
     5161 gatttagagg agttagtctt gaagtcatgc gccggttaag gctaaactga aaggacaagt
     5221 tttggtgact gcgctcctcc aagccagtta cctcggttca aagagttggt agctcagaga
     5281 accttcgaaa aaccgccctg caaggcggtt ttttcgtttt cagagcaaga gattacgcgc
     5341 agaccaaaac gatctcaaga agatcatctt attaaggggt ctgacgctca gtggaacgaa
     5401 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     5461 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     5521 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     5581 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     5641 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     5701 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     5761 cagtctattc catggtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     5821 ataaaaatag gcgtatcacg aggcagaatt tcagataaaa aaaatcctta gctttcgcta
     5881 aggatgattt ctggaattcg cggccgcttc tagag
//
