LOCUS       sequence-426111-        4740 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  biphasic target plasmid version 2.0 (""flip-proof"").
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4740)
  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 4740)
  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: pDSG588
FEATURES             Location/Qualifiers
     source          1..4740
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..35
                     /label=BBa_J23116
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.16"
     RBS             101..112
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             119..355
                     /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    2078..2098
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      2099..2156
                     /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(2252..3043,3044..3112))
                     /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     2875..2894
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3113..3184)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3638..4183)
                     /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(3718..3737)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4651..4669
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(4673..4716)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    4719..4740
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 ttgacagcta gctcagtcct agggactatg ctagctacta gagaactttt cggatcaagc
       61 tatgaaggac gcaaagaggg aactaaacac tttactagag aaagaggaga aatactagat
      121 gggtcaaagt agcgatgaag ccaacgctcc cgttgcaggg cagtttgcgc ttcccctgag
      181 tgccaccttt ggcttagggg atcgcgtacg caagaaatct ggtgccgctt ggcagggtca
      241 agtcgtcggt tggtattgca caaaactcac tcctgaaggc tatgcggtcg agtccgaatc
      301 ccacccaggc tcagtgcaaa tttatcctgt ggctgcactt gaacgtgtgg cctaataact
      361 cggtaccaaa ttccagaaaa gaggcctccc gaaagggggg ccttttttcg ttttggtccg
      421 catccacctg tgcggacctc ggtaccaaat tccagaaaag agacgctttc gagcgtcttt
      481 tttcgttttg gtccatcgag tggcgcgctt agttcctcgt tttctctcgt tggaagaaga
      541 agaaacgaga aactaaatac tagagtcaca caggactact agatgatgca aaccgtttta
      601 gcgaaaatcg tcgcagacaa ggcgatttgg gtagaagccc gcaaacagca gcaaccgctg
      661 gccagttttc agaatgaggt tcagccgagc acgcgacatt tttatgatgc gctacagggt
      721 gcgcgcacgg cgtttattct ggagtgcaag aaagcgtcgc cgtcaaaagg cgtgatccgt
      781 gatgatttcg atccagcacg cattgccgcc atttataaac attacgcttc ggcaatttcg
      841 gtgctgactg atgagaaata ttttcagggg agctttaatt tcctccccat cgtcagccaa
      901 atcgccccgc agccgatttt atgtaaagac ttcattatcg acccttacca gatctatctg
      961 gcgcgctatt accaggccga tgcctgctta ttaatgcttt cagtactgga tgacgaccaa
     1021 tatcgccagc ttgccgccgt cgctcacagt ctggagatgg gggtgctgac cgaagtcagt
     1081 aatgaagagg aacaggagcg cgccattgca ttgggagcaa aggtcgttgg catcaacaac
     1141 cgcgatctgc gtgatttgtc gattgatctc aaccgtaccc gcgagcttgc gccgaaactg
     1201 gggcacaacg tgacggtaat cagcgaatcc ggcatcaata cttacgctca ggtgcgcgag
     1261 ttaagccact tcgctaacgg ttttctgatt ggttcggcgt tgatggccca tgacgatttg
     1321 cacgccgccg tgcgccgggt gttgctgggt gagaataaag tatgtggcct gacgcgtggg
     1381 caagatgcta aagcagctta tgacgcgggc gcgatttacg gtgggttgat ttttgttgcg
     1441 acatcaccgc gttgcgtcaa cgttgaacag gcgcaggaag tgatggctgc ggcaccgttg
     1501 cagtatgttg gcgtgttccg caatcacgat attgccgatg tggtggacaa agctaaggtg
     1561 ttatcgctgg cggcagtgca actgcatggt aatgaagaac agctgtatat cgatacgctg
     1621 cgtgaagctc tgccagcaca tgttgccatc tggaaagcat taagcgtcgg tgaaaccctg
     1681 cccgcccgcg agtttcagca cgttgataaa tatgttttag acaacggcca gggtggaagc
     1741 gggcaacgtt ttgactggtc actattaaat ggtcaatcgc ttggcaacgt tctgctggcg
     1801 gggggcttag gcgcagataa ctgcgtggaa gcggcacaaa ccggctgcgc cggacttgat
     1861 tttaattctg ctgtagagtc gcaaccgggc atcaaagacg cacgtctttt ggcctcggtt
     1921 ttccagacgc tgcgcgcata ttaataactc ggtaccaaag acgaacaata agacgctgaa
     1981 aagcgtcttt tttcgttttg gtcctttgaa gtttagggga tagtccaatt attgaaggcc
     2041 tccctaacgg ggggcctttt tttgtttctg gtctccctac tagtagcggc cgctgcagtc
     2101 cggcaaaaaa gggcaaggtg tcaccaccct gccctttttc tttaaaaccg aaaagattac
     2161 ttcgcgttat gcaggcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     2221 gagcggtatc agctcactca aaggcggtaa tttaccaatg cttaatcagt gaggcaccta
     2281 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     2341 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     2401 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     2461 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     2521 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     2581 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     2641 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     2701 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     2761 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     2821 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     2881 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     2941 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     3001 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     3061 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     3121 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     3181 aatgtattta gatcacgcat cttcccgaca acgcagaccg ttccgtggca aagcaaaagt
     3241 tcaaaatcac caactggtcc acctacaaca aagctctcat caaccgtggc tccctcactt
     3301 tctggctgga tgatggggcg attcaggcct ggtatgagtc agcaacacct tcttcacgag
     3361 gcagacctca gcgctagcgg agtgtatact ggcttactat gttggcactg atgagggtgt
     3421 cagtgaagtg cttcatgtgg caggagaaaa aaggctgcac cggtgcgtca gcagaatatg
     3481 tgatacagga tatattccgc ttcctcgctc actgactcgc tacgctcggt cgttcgactg
     3541 cggcgagcgg aaatggctta cgaacggggc ggagatttcc tggaagatgc caggaagata
     3601 cttaacaggg aagtgagagg gccgcggcaa agccgttttt ccataggctc cgcccccctg
     3661 acaagcatca cgaaatctga cgctcaaatc agtggtggcg aaacccgaca ggactataaa
     3721 gataccaggc gtttccccct ggcggctccc tcgtgcgctc tcctgttcct gcctttcggt
     3781 ttaccggtgt cattccgctg ttatggccgc gtttgtctca ttccacgcct gacactcagt
     3841 tccgggtagg cagttcgctc caagctggac tgtatgcacg aaccccccgt tcagtccgac
     3901 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggaaagaca tgcaaaagca
     3961 ccactggcag cagccactgg taattgattt agaggagtta gtcttgaagt catgcgccgg
     4021 ttaaggctaa actgaaagga caagttttgg tgactgcgct cctccaagcc agttacctcg
     4081 gttcaaagag ttggtagctc agagaacctt cgaaaaaccg ccctgcaagg cggttttttc
     4141 gttttcagag caagagatta cgcgcagacc aaaacgatct caagaagatc atcttattaa
     4201 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     4261 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     4321 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     4381 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     4441 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     4501 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     4561 tcctgcaact ttatccgcct ccatccagtc tattccatgg tgccacctga cgtctaagaa
     4621 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggca gaatttcaga
     4681 taaaaaaaat ccttagcttt cgctaaggat gatttctgga attcgcggcc gcttctagag
//
