LOCUS       sequence-251917-        4874 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 4874)
  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 4874)
  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: pDD33
FEATURES             Location/Qualifiers
     source          1..4874
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(154..156,157..159,160..867)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     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"
     CDS             907..1878
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     terminator      2023..2050
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(2181..2972,2973..3041))
                     /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     2804..2823
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3042..3146)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3567..3584)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3701..4246)
                     /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(3781..3800)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4322..4341
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     protein_bind    4499..4515
                     /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     4504..4526
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             4717..4746
                     /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 gctgtacggc ggtggctctg gaggtggtgg gtccggcggt
      901 ggctctatga agcgtgcgcc tgttattcca aaacatacgc tcaatactca accggttgaa
      961 gatacttcgt tatcgacacc agctgccccg atggtggatt cgttaattgc gcgcgtagga
     1021 gtaatggctc gcggtaatgc cattactttg cctgtatgtg gtcgggatgt gaagtttact
     1081 cttgaagtgc tccggggtga tagtgttgag aagacctctc gggtatggtc aggtaatgaa
     1141 cgtgaccagg agctgcttac tgaggacgca ctggatgatc tcatcccttc ttttctactg
     1201 actggtcaac agacaccggc gttcggtcga agagtatctg gtgtcataga aattgccgat
     1261 gggagtcgcc gtcgtaaagc tgctgcactt accgaaagtg attatcgtgt tctggttggc
     1321 gagctggatg atgagcagat ggctgcatta tccagattgg gtaacgatta tcgcccaaca
     1381 agtgcttatg aacgtggtca gcgttatgca agccgattgc agaatgaatt tgctggaaat
     1441 atttctgcgc tggctgatgc ggaaaatatt tcacgtaaga ttattacccg ctgtatcaac
     1501 accgccaaat tgcctaaatc agttgttgct cttttttctc accccggtga actatctgcc
     1561 cggtcaggtg atgcacttca aaaagccttt acagataaag aggaattact taagcagcag
     1621 gcatctaacc ttcatgagca gaaaaaagct ggggtgatat ttgaagctga agaagttatc
     1681 actcttttaa cttctgtgct taaaacgtca tctgcatcaa gaactagttt aagctcacga
     1741 catcagtttg ctcctggagc gacagtattg tataagggcg ataaaatggt gcttaacctg
     1801 gacaggtctc gtgttccaac tgagtgtata gagaaaattg aggccattct taaggaactt
     1861 gaaaagccag caccctgaaa gcttgctacg tgctgctgaa gttgcccgca acagagagtg
     1921 gaaccaaccg gtgataccac gatactatga ctgagagtca acgccatgag cggcctcatt
     1981 tcttattctg agttacaaca gtccgcaccg ctgtctcaca ctggctcacc ttcgggtggg
     2041 cctttctgcg tttatatact agagagagaa tataaaaagc cagattatta atccggcttt
     2101 tttattattt cctagctagc cgcacttatt caggcgtagc aaccaggcgt ttaagggcac
     2161 caataactgc cttaaaaaaa ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     2221 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     2281 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     2341 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     2401 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     2461 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     2521 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     2581 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     2641 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     2701 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     2761 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     2821 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     2881 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     2941 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     3001 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     3061 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     3121 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgtc
     3181 tcattttcgc caaaagttgg cccagggctt cccggtatca acagggacac cagaatttat
     3241 ttattctgcg aagtgatctt ccgtcacagg tatttattcg gcgcaaagtg cgtcgggtga
     3301 tgctgccaac ttactgattt agtgtatgat ggtgtttttg aggtgctcca gtggcttctg
     3361 tttctatcag ctgtccctcc tgttcagcta ctgacggggt ggtgcgtaac ggcaaaagca
     3421 ccgccggaca tcagcgctag cggagtgtat actggcttac tatgttggca ctgatgaggg
     3481 tgtcagtgaa gtgcttcatg tggcaggaga aaaaaggctg caccggtgcg tcagcagaat
     3541 atgtgataca ggatatattc cgcttcctcg ctcactgact cgctacgctc ggtcgttcga
     3601 ctgcggcgag cggaaatggc ttacgaacgg ggcggagatt tcctggaaga tgccaggaag
     3661 atacttaaca gggaagtgag agggccgcgg caaagccgtt tttccatagg ctccgccccc
     3721 ctgacaagca tcacgaaatc tgacgctcaa atcagtggtg gcgaaacccg acaggactat
     3781 aaagatacca ggcgtttccc cctggcggct ccctcgtgcg ctctcctgtt cctgcctttc
     3841 ggtttaccgg tgtcattccg ctgttatggc cgcgtttgtc tcattccacg cctgacactc
     3901 agttccgggt aggcagttcg ctccaagctg gactgtatgc acgaaccccc cgttcagtcc
     3961 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggaaag acatgcaaaa
     4021 gcaccactgg cagcagccac tggtaattga tttagaggag ttagtcttga agtcatgcgc
     4081 cggttaaggc taaactgaaa ggacaagttt tggtgactgc gctcctccaa gccagttacc
     4141 tcggttcaaa gagttggtag ctcagagaac cttcgaaaaa ccgccctgca aggcggtttt
     4201 ttcgttttca gagcaagaga ttacgcgcag accaaaacga tctcaagaag atcatcttat
     4261 taatcagata aaatatttct aggctcatga gcccgaagtg gcgagcccga tcttccccat
     4321 cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca
     4381 cgatgcgtcc ggcgtagagg atctgctcat gtttgacagc ttatcgagct cagtgcgctc
     4441 gagcttccca accttaccag agggcgcccc agctggcaat tccgacgtct gtgtggaatt
     4501 gtgagcggat aacaatttca cacagggccc tcggacaccg aggagaatgt caagaggcga
     4561 acacacaacg tcttggagcg ccagaggagg aacgagctaa aacggagctt ttttgccctg
     4621 cgtgaccaga tcccggagtt ggaaaacaat gaaaaggccc ccaaggtagt tatccttaaa
     4681 aaagccacag catacatcct gtccgtccaa gcagaggagc aaaagctcat ttctgaagag
     4741 gacttgttgc ggaaacgacg agaacagttg aaacacaaac ttgaacagct acggaactct
     4801 tgtgcgtaag gaaaagtaag gaaaacgatt ccttctaaca gaaatgtcct gagcaatcac
     4861 ctatgaactg tcga
//
