LOCUS       sequence-251975-        4805 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  asymmetric plasmid partitioning E. coli - regulatory plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4805)
  AUTHORS   Molinari S, Shis DL, Bhakta SP, Chappell J, Igoshin OA, Bennett MR
  TITLE     A synthetic system for asymmetric cell division in Escherichia coli.
  JOURNAL   Nat Chem Biol. 2019 Sep;15(9):917-924. doi: 
            10.1038/s41589-019-0339-x. Epub 2019 Aug 12.
  PUBMED    31406375
REFERENCE   2  (bases 1 to 4805)
  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: 
            "10.1038/s41589-019-0339-x"; journalName: "Nat Chem Biol"; date: 
            "2019-09"; volume: "15"; issue: "9"; pages: "917-924"
COMMENT     Sequence Label: pSM019
FEATURES             Location/Qualifiers
     source          1..4805
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             154..867
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     misc_feature    868..924
                     /label=MCS
                     /note="multiple cloning site"
     terminator      1954..1981
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(2112..2903,2904..2972))
                     /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     2735..2754
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2973..3077)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3498..3515)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3632..4177)
                     /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(3712..3731)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4253..4272
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     protein_bind    4430..4446
                     /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)."
     primer_bind     4435..4457
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             4648..4677
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
ORIGIN
        1 catagcattt ttatccataa gattagcgga tcctaccttt tacttcttgc catcgactac
       61 ggatactgag cacatgcatg ccacttaaaa aggagatcaa caatgaaagc aattttcgta
      121 ctgaaacatc ttaatcatgc taaggagact agtatgcgta aaggcgaaga gctgttcact
      181 ggtgtcgtcc ctattctggt ggaactggat ggtgatgtca acggtcataa gttttccgtg
      241 cgtggcgagg gtgaaggtga cgcaactaat ggtaaactga cgctgaagtt catctgtact
      301 actggtaaac tgccggtacc ttggccgact ctggtaacga cgctgactta tggtgttcag
      361 tgctttgctc gttatccgga ccatatgaag cagcatgact tcttcaagtc cgccatgccg
      421 gaaggctatg tgcaggaacg cacgatttcc tttaaggatg acggcacgta caaaacgcgt
      481 gcggaagtga aatttgaagg cgataccctg gtaaaccgca ttgagctgaa aggcattgac
      541 tttaaagaag acggcaatat cctgggccat aagctggaat acaattttaa cagccacaat
      601 gtttacatca ccgccgataa acaaaaaaat ggcattaaag cgaattttaa aattcgccac
      661 aacgtggagg atggcagcgt gcagctggct gatcactacc agcaaaacac tccaatcggt
      721 gatggtcctg ttctgctgcc agacaatcac tatctgagct accaaagcgt tctgtctaaa
      781 gatccgaacg agaaacgcga tcatatggtt ctgctggagt tcgtaaccgc agcgggcatc
      841 acgcatggta tggatgaact gtacaagaga tctcgagctc aagcttcgaa ttctgcagtc
      901 gacggtaccg cgggcccggg atccatgtcc gaagggcgtc gtggtctggg tcgagggctt
      961 tcggccctgc tgggcgaggt cgacgccgcg ccggctcagg cgccgggcga gcagcttggc
     1021 ggttcccgtg aagcgccgat cgaaatcctg cagcgcaatc ccgaccagcc ccgtcggacc
     1081 ttccgcgagg aagacctcga ggatctgtca aactcgatcc gtgagaaggg cgttctgcag
     1141 ccgatcctgg tgcggccgtc gccggatacg gctggcgagt accagatcgt cgccggcgag
     1201 cgccgttggc gcgcagccca gcgggcgggc ctcaagactg tgccgatcat ggtgcgcgag
     1261 ctcgacgacc tggcggtgct ggagatcggc atcatcgaga acgtccagcg ggctgatctg
     1321 aacgtcctgg aagaagcgct ctcctacaag gtgctgatgg agaagttcga gcgcacccag
     1381 gagaacatcg cccagaccat cggcaagagc cgcagccacg tggccaacac gatgcgcttg
     1441 ctggccctac cggacgaggt tcagtcgcac ctggtcagtg gtgagctgac ggccggccac
     1501 gcccgcgcca tcgctgcagc ggctgacccg gtggccctcg ccaagcaaat catcgagggc
     1561 ggcttgtcgg tgcgcgagac cgaggccctg gcccgtaagg cgcccaacct gtctgcgggc
     1621 aagagcaagg ggggccgccc cccacgcgtg aaggacaccg acacccaggc gctggagtct
     1681 gacctgtcct cggttctggg tctggatgtc tcgatcgacc atcgcgggtc taccggcacg
     1741 ctgaccatca cctacgccac cctcgagcag ctcgacgacc tctgcaaccg actgacgcgc
     1801 gggatctgaa agcttgctac gtgctgctga agttgcccgc aacagagagt ggaaccaacc
     1861 ggtgatacca cgatactatg actgagagtc aacgccatga gcggcctcat ttcttattct
     1921 gagttacaac agtccgcacc gctgtctcac actggctcac cttcgggtgg gcctttctgc
     1981 gtttatatac tagagagaga atataaaaag ccagattatt aatccggctt ttttattatt
     2041 tcctagctag ccgcacttat tcaggcgtag caaccaggcg tttaagggca ccaataactg
     2101 ccttaaaaaa attaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     2161 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     2221 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     2281 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     2341 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     2401 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     2461 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     2521 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     2581 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     2641 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     2701 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     2761 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     2821 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     2881 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     2941 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     3001 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     3061 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcgt ctcattttcg
     3121 ccaaaagttg gcccagggct tcccggtatc aacagggaca ccagaattta tttattctgc
     3181 gaagtgatct tccgtcacag gtatttattc ggcgcaaagt gcgtcgggtg atgctgccaa
     3241 cttactgatt tagtgtatga tggtgttttt gaggtgctcc agtggcttct gtttctatca
     3301 gctgtccctc ctgttcagct actgacgggg tggtgcgtaa cggcaaaagc accgccggac
     3361 atcagcgcta gcggagtgta tactggctta ctatgttggc actgatgagg gtgtcagtga
     3421 agtgcttcat gtggcaggag aaaaaaggct gcaccggtgc gtcagcagaa tatgtgatac
     3481 aggatatatt ccgcttcctc gctcactgac tcgctacgct cggtcgttcg actgcggcga
     3541 gcggaaatgg cttacgaacg gggcggagat ttcctggaag atgccaggaa gatacttaac
     3601 agggaagtga gagggccgcg gcaaagccgt ttttccatag gctccgcccc cctgacaagc
     3661 atcacgaaat ctgacgctca aatcagtggt ggcgaaaccc gacaggacta taaagatacc
     3721 aggcgtttcc ccctggcggc tccctcgtgc gctctcctgt tcctgccttt cggtttaccg
     3781 gtgtcattcc gctgttatgg ccgcgtttgt ctcattccac gcctgacact cagttccggg
     3841 taggcagttc gctccaagct ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc
     3901 gccttatccg gtaactatcg tcttgagtcc aacccggaaa gacatgcaaa agcaccactg
     3961 gcagcagcca ctggtaattg atttagagga gttagtcttg aagtcatgcg ccggttaagg
     4021 ctaaactgaa aggacaagtt ttggtgactg cgctcctcca agccagttac ctcggttcaa
     4081 agagttggta gctcagagaa ccttcgaaaa accgccctgc aaggcggttt tttcgttttc
     4141 agagcaagag attacgcgca gaccaaaacg atctcaagaa gatcatctta ttaatcagat
     4201 aaaatatttc taggctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt
     4261 cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc
     4321 cggcgtagag gatctgctca tgtttgacag cttatcgagc tcagtgcgct cgagcttccc
     4381 aaccttacca gagggcgccc cagctggcaa ttccgacgtc tgtgtggaat tgtgagcgga
     4441 taacaatttc acacagggcc ctcggacacc gaggagaatg tcaagaggcg aacacacaac
     4501 gtcttggagc gccagaggag gaacgagcta aaacggagct tttttgccct gcgtgaccag
     4561 atcccggagt tggaaaacaa tgaaaaggcc cccaaggtag ttatccttaa aaaagccaca
     4621 gcatacatcc tgtccgtcca agcagaggag caaaagctca tttctgaaga ggacttgttg
     4681 cggaaacgac gagaacagtt gaaacacaaa cttgaacagc tacggaactc ttgtgcgtaa
     4741 ggaaaagtaa ggaaaacgat tccttctaac agaaatgtcc tgagcaatca cctatgaact
     4801 gtcga
//
