LOCUS       sequence-251664-        4336 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 4336)
  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 4336)
  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: pSPB866
FEATURES             Location/Qualifiers
     source          1..4336
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             153..161
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             198..254
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             join(315..317,318..1028)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(357..378)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(618..637)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     965..986
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1029..1085
                     /label=MCS
                     /note="multiple cloning site"
     misc_feature    2050..2070
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     CDS             complement(join(2298..3089,3090..3158))
                     /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     2921..2940
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3159..3263)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(3307..3338)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     primer_bind     complement(3482..3499)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3616..4161)
                     /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(3696..3715)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             4228..4239
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_feature    4286..4307
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 cgcacccata gcatttttat ccataagatt agcggatcct accttttact tcttgccatc
       61 gactacggat actgagcacg acgggcccaa gttcacttaa aaaggagatc aacaatgaaa
      121 gcaattttcg tactgaaaca tcttaatcat gctaaggagg ttttctaatg tcggttaagg
      181 aactggaaga taaagtcgaa gaactgctga gcaaaaacta tcatctggaa aacgaagtgg
      241 cgcgcctgaa aaaactggtg ggcgaacgca agagaggcgg tggctctgga ggtggtgggt
      301 ccggcggtgg ttccgtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
      361 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
      421 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
      481 cctggcccac cctcgtgacc accctgacct ggggcgtgca gtgcttcagc cgctaccccg
      541 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
      601 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
      661 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
      721 tcctggggca caagctggag tacaactaca tcagccacaa cgtctatatc accgccgaca
      781 agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag gacggcagcg
      841 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
      901 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
      961 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     1021 tgtacaagag atctcgagct caagcttcga attctgcagt cgacggtacc gcgggcccgg
     1081 gatccatgtc cgaagggcgt cgtggtctgg gtcgagggct ttcggccctg ctgggcgagg
     1141 tcgacgccgc gccggctcag gcgccgggcg agcagcttgg cggttcccgt gaagcgccga
     1201 tcgaaatcct gcagcgcaat cccgaccagc cccgtcggac cttccgcgag gaagacctcg
     1261 aggatctgtc aaactcgatc cgtgagaagg gcgttctgca gccgatcctg gtgcggccgt
     1321 cgccggatac ggctggcgag taccagatcg tcgccggcga gcgccgttgg cgcgcagccc
     1381 agcgggcggg cctcaagact gtgccgatca tggtgcgcga gctcgacgac ctggcggtgc
     1441 tggagatcgg catcatcgag aacgtccagc gggctgatct gaacgtcctg gaagaagcgc
     1501 tctcctacaa ggtgctgatg gagaagttcg agcgcaccca ggagaacatc gcccagacca
     1561 tcggcaagag ccgcagccac gtggccaaca cgatgcgctt gctggcccta ccggacgagg
     1621 ttcagtcgca cctggtcagt ggtgagctga cggccggcca cgcccgcgcc atcgctgcag
     1681 cggctgaccc ggtggccctc gccaagcaaa tcatcgaggg cggcttgtcg gtgcgcgaga
     1741 cagaggccct ggcccgtaag gcgcccaacc tgtctgcggg caagagcaag gggggccgcc
     1801 ccccacgcgt gaaggacacc gacacccagg cgctggagtc tgacctgtcc tcggttctgg
     1861 gtctggatgt ctcgatcgac catcgcgggt ctaccggcac gctgaccatc acctacgcca
     1921 ccctcgagca gctcgacgac ctctgcaacc gactgacgcg cgggatctaa taatggcctc
     1981 ggtaccaaag acgaacaata agacgctgaa aagcgtcttt tttcgttttg gtccgctgcg
     2041 tttgagacgt actagtagcg gccgctgcag ttaatcactg attaactcgg taccaaattc
     2101 cagaaaagag gcctcccgaa aggggggcct tttttcgttt tggtcctttt gttatcaata
     2161 aaaaagggga gcggtttccc gctcccctta ttgttcgtct acagattatc aaaaaggatc
     2221 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     2281 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     2341 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     2401 ggcttaccat ctggccccag tgctgcaatg ataccgcggg acccacgctc accggctcca
     2461 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     2521 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     2581 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     2641 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     2701 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     2761 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     2821 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     2881 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     2941 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     3001 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     3061 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     3121 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     3181 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     3241 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctgg agtcctgttg
     3301 agtaatagtc aaaagcctcc ggtcggaggc ttttgacttt ctgcttaccc aatagcggag
     3361 tgtatactgg cttactatgt tggcactgat gagggtgtca gtgaagtgct tcatgtggca
     3421 ggagaaaaaa ggctgcaccg gtgcgtcagc agaatatgtg atacaggata tattccgctt
     3481 cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg gcgagcggaa atggcttacg
     3541 aacggggcgg agatttcctg gaagatgcca ggaagatact taacagggaa gtgagagggc
     3601 cgcggcaaag ccgtttttcc ataggctccg cccccctgac aagcatcacg aaatctgacg
     3661 ctcaaatcag tggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3721 cggctccctc gtgcgctctc ctgttcctgc ctttcggttt accggtgtca ttccgctgtt
     3781 atggccgcgt ttgtctcatt ccacgcctga cactcagttc cgggtaggca gttcgctcca
     3841 agctggactg tatgcacgaa ccccccgttc agtccgaccg ctgcgcctta tccggtaact
     3901 atcgtcttga gtccaacccg gaaagacatg caaaagcacc actggcagca gccactggta
     3961 attgatttag aggagttagt cttgaagtca tgcgccggtt aaggctaaac tgaaaggaca
     4021 agttttggtg actgcgctcc tccaagccag ttacctcggt tcaaagagtt ggtagctcag
     4081 agaaccttcg aaaaaccgcc ctgcaaggcg gttttttcgt tttcagagca agagattacg
     4141 cgcagaccaa aacgatctca agaagatcat cttattaaac ggaaaaaaaa accccgcccc
     4201 tgacagggcg gggttttttt tccaattatt gaaggccgct aacgcggcct ttttttgttt
     4261 ctggtctgcc ttaatcaatg actaagaatt cgcggccgct tctagagcgt ctcacttcgg
     4321 atccgctgca ggcttc
//
