LOCUS       sequence-251630-        4808 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 4808)
  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 4808)
  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: pSM015
FEATURES             Location/Qualifiers
     source          1..4808
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(154..156,157..159,160..870)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(199..220)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(460..479)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     807..828
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    871..927
                     /label=MCS
                     /note="multiple cloning site"
     terminator      1957..1984
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(2115..2906,2907..2975))
                     /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     2738..2757
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2976..3080)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3501..3518)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3635..4180)
                     /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(3715..3734)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4256..4275
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     protein_bind    4433..4449
                     /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     4438..4460
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             4651..4680
                     /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 agtatggtga gcaagggcga ggagctgttc
      181 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
      241 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
      301 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctggggcgtg
      361 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
      421 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
      481 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
      541 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta catcagccac
      601 aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc
      661 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
      721 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
      781 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
      841 atcactctcg gcatggacga gctgtacaag agatctcgag ctcaagcttc gaattctgca
      901 gtcgacggta ccgcgggccc gggatccatg tccgaagggc gtcgtggtct gggtcgaggg
      961 ctttcggccc tgctgggcga ggtcgacgcc gcgccggctc aggcgccggg cgagcagctt
     1021 ggcggttccc gtgaagcgcc gatcgaaatc ctgcagcgca atcccgacca gccccgtcgg
     1081 accttccgcg aggaagacct cgaggatctg tcaaactcga tccgtgagaa gggcgttctg
     1141 cagccgatcc tggtgcggcc gtcgccggat acggctggcg agtaccagat cgtcgccggc
     1201 gagcgccgtt ggcgcgcagc ccagcgggcg ggcctcaaga ctgtgccgat catggtgcgc
     1261 gagctcgacg acctggcggt gctggagatc ggcatcatcg agaacgtcca gcgggctgat
     1321 ctgaacgtcc tggaagaagc gctctcctac aaggtgctga tggagaagtt cgagcgcacc
     1381 caggagaaca tcgcccagac catcggcaag agccgcagcc acgtggccaa cacgatgcgc
     1441 ttgctggccc taccggacga ggttcagtcg cacctggtca gtggtgagct gacggccggc
     1501 cacgcccgcg ccatcgctgc agcggctgac ccggtggccc tcgccaagca aatcatcgag
     1561 ggcggcttgt cggtgcgcga gaccgaggcc ctggcccgta aggcgcccaa cctgtctgcg
     1621 ggcaagagca aggggggccg ccccccacgc gtgaaggaca ccgacaccca ggcgctggag
     1681 tctgacctgt cctcggttct gggtctggat gtctcgatcg accatcgcgg gtctaccggc
     1741 acgctgacca tcacctacgc caccctcgag cagctcgacg acctctgcaa ccgactgacg
     1801 cgcgggatct gaaagcttgc tacgtgctgc tgaagttgcc cgcaacagag agtggaacca
     1861 accggtgata ccacgatact atgactgaga gtcaacgcca tgagcggcct catttcttat
     1921 tctgagttac aacagtccgc accgctgtct cacactggct caccttcggg tgggcctttc
     1981 tgcgtttata tactagagag agaatataaa aagccagatt attaatccgg cttttttatt
     2041 atttcctagc tagccgcact tattcaggcg tagcaaccag gcgtttaagg gcaccaataa
     2101 ctgccttaaa aaaattacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     2161 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     2221 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     2281 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     2341 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     2401 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     2461 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     2521 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     2581 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     2641 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     2701 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     2761 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     2821 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     2881 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     2941 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     3001 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     3061 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgtctcattt
     3121 tcgccaaaag ttggcccagg gcttcccggt atcaacaggg acaccagaat ttatttattc
     3181 tgcgaagtga tcttccgtca caggtattta ttcggcgcaa agtgcgtcgg gtgatgctgc
     3241 caacttactg atttagtgta tgatggtgtt tttgaggtgc tccagtggct tctgtttcta
     3301 tcagctgtcc ctcctgttca gctactgacg gggtggtgcg taacggcaaa agcaccgccg
     3361 gacatcagcg ctagcggagt gtatactggc ttactatgtt ggcactgatg agggtgtcag
     3421 tgaagtgctt catgtggcag gagaaaaaag gctgcaccgg tgcgtcagca gaatatgtga
     3481 tacaggatat attccgcttc ctcgctcact gactcgctac gctcggtcgt tcgactgcgg
     3541 cgagcggaaa tggcttacga acggggcgga gatttcctgg aagatgccag gaagatactt
     3601 aacagggaag tgagagggcc gcggcaaagc cgtttttcca taggctccgc ccccctgaca
     3661 agcatcacga aatctgacgc tcaaatcagt ggtggcgaaa cccgacagga ctataaagat
     3721 accaggcgtt tccccctggc ggctccctcg tgcgctctcc tgttcctgcc tttcggttta
     3781 ccggtgtcat tccgctgtta tggccgcgtt tgtctcattc cacgcctgac actcagttcc
     3841 gggtaggcag ttcgctccaa gctggactgt atgcacgaac cccccgttca gtccgaccgc
     3901 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg aaagacatgc aaaagcacca
     3961 ctggcagcag ccactggtaa ttgatttaga ggagttagtc ttgaagtcat gcgccggtta
     4021 aggctaaact gaaaggacaa gttttggtga ctgcgctcct ccaagccagt tacctcggtt
     4081 caaagagttg gtagctcaga gaaccttcga aaaaccgccc tgcaaggcgg ttttttcgtt
     4141 ttcagagcaa gagattacgc gcagaccaaa acgatctcaa gaagatcatc ttattaatca
     4201 gataaaatat ttctaggctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga
     4261 tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc
     4321 gtccggcgta gaggatctgc tcatgtttga cagcttatcg agctcagtgc gctcgagctt
     4381 cccaacctta ccagagggcg ccccagctgg caattccgac gtctgtgtgg aattgtgagc
     4441 ggataacaat ttcacacagg gccctcggac accgaggaga atgtcaagag gcgaacacac
     4501 aacgtcttgg agcgccagag gaggaacgag ctaaaacgga gcttttttgc cctgcgtgac
     4561 cagatcccgg agttggaaaa caatgaaaag gcccccaagg tagttatcct taaaaaagcc
     4621 acagcataca tcctgtccgt ccaagcagag gagcaaaagc tcatttctga agaggacttg
     4681 ttgcggaaac gacgagaaca gttgaaacac aaacttgaac agctacggaa ctcttgtgcg
     4741 taaggaaaag taaggaaaac gattccttct aacagaaatg tcctgagcaa tcacctatga
     4801 actgtcga
//
