LOCUS       sequence-251335-        3625 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 3625)
  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 3625)
  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: pSPB858
FEATURES             Location/Qualifiers
     source          1..3625
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             50..61
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_feature    108..129
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     RBS             311..319
                     /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             356..412
                     /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"
     misc_feature    476..532
                     /label=MCS
                     /note="multiple cloning site"
     misc_feature    1497..1517
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     CDS             complement(join(1745..2536,2537..2605))
                     /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     2368..2387
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2606..2710)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(2754..2785)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     primer_bind     complement(2929..2946)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3063..3608)
                     /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(3143..3162)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acggaaaaaa aaaccccgcc cctgacaggg cggggttttt tttccaatta ttgaaggccg
       61 ctaacgcggc ctttttttgt ttctggtctg ccttaatcaa tgactaagaa ttcgcggccg
      121 cttctagagc gtctcacttc ggatccgctg caggcttccg cacccatagc atttttatcc
      181 ataagattag cggatcctac cttttacttc ttgccatcga ctacggatac tgagcacgac
      241 gggcccaagt tcacttaaaa aggagatcaa caatgaaagc aattttcgta ctgaaacatc
      301 ttaatcatgc taaggaggtt ttctaatgtc ggttaaggaa ctggaagata aagtcgaaga
      361 actgctgagc aaaaactatc atctggaaaa cgaagtggcg cgcctgaaaa aactggtggg
      421 cgaacgcaag agaggcggtg gctctggagg tggtgggtcc ggcggtggtt ccaagagatc
      481 tcgagctcaa gcttcgaatt ctgcagtcga cggtaccgcg ggcccgggat ccatgtccga
      541 agggcgtcgt ggtctgggtc gagggctttc ggccctgctg ggcgaggtcg acgccgcgcc
      601 ggctcaggcg ccgggcgagc agcttggcgg ttcccgtgaa gcgccgatcg aaatcctgca
      661 gcgcaatccc gaccagcccc gtcggacctt ccgcgaggaa gacctcgagg atctgtcaaa
      721 ctcgatccgt gagaagggcg ttctgcagcc gatcctggtg cggccgtcgc cggatacggc
      781 tggcgagtac cagatcgtcg ccggcgagcg ccgttggcgc gcagcccagc gggcgggcct
      841 caagactgtg ccgatcatgg tgcgcgagct cgacgacctg gcggtgctgg agatcggcat
      901 catcgagaac gtccagcggg ctgatctgaa cgtcctggaa gaagcgctct cctacaaggt
      961 gctgatggag aagttcgagc gcacccagga gaacatcgcc cagaccatcg gcaagagccg
     1021 cagccacgtg gccaacacga tgcgcttgct ggccctaccg gacgaggttc agtcgcacct
     1081 ggtcagtggt gagctgacgg ccggccacgc ccgcgccatc gctgcagcgg ctgacccggt
     1141 ggccctcgcc aagcaaatca tcgagggcgg cttgtcggtg cgcgagacag aggccctggc
     1201 ccgtaaggcg cccaacctgt ctgcgggcaa gagcaagggg ggccgccccc cacgcgtgaa
     1261 ggacaccgac acccaggcgc tggagtctga cctgtcctcg gttctgggtc tggatgtctc
     1321 gatcgaccat cgcgggtcta ccggcacgct gaccatcacc tacgccaccc tcgagcagct
     1381 cgacgacctc tgcaaccgac tgacgcgcgg gatctaataa tggcctcggt accaaagacg
     1441 aacaataaga cgctgaaaag cgtctttttt cgttttggtc cgctgcgttt gagacgtact
     1501 agtagcggcc gctgcagtta atcactgatt aactcggtac caaattccag aaaagaggcc
     1561 tcccgaaagg ggggcctttt ttcgttttgg tccttttgtt atcaataaaa aaggggagcg
     1621 gtttcccgct ccccttattg ttcgtctaca gattatcaaa aaggatcttc acctagatcc
     1681 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     1741 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     1801 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     1861 gccccagtgc tgcaatgata ccgcgggacc cacgctcacc ggctccagat ttatcagcaa
     1921 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     1981 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     2041 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     2101 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     2161 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     2221 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     2281 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     2341 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     2401 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     2461 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     2521 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     2581 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     2641 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     2701 gggttccgcg cacatttccc cgaaaagtgc cacctggagt cctgttgagt aatagtcaaa
     2761 agcctccggt cggaggcttt tgactttctg cttacccaat agcggagtgt atactggctt
     2821 actatgttgg cactgatgag ggtgtcagtg aagtgcttca tgtggcagga gaaaaaaggc
     2881 tgcaccggtg cgtcagcaga atatgtgata caggatatat tccgcttcct cgctcactga
     2941 ctcgctacgc tcggtcgttc gactgcggcg agcggaaatg gcttacgaac ggggcggaga
     3001 tttcctggaa gatgccagga agatacttaa cagggaagtg agagggccgc ggcaaagccg
     3061 tttttccata ggctccgccc ccctgacaag catcacgaaa tctgacgctc aaatcagtgg
     3121 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggcgg ctccctcgtg
     3181 cgctctcctg ttcctgcctt tcggtttacc ggtgtcattc cgctgttatg gccgcgtttg
     3241 tctcattcca cgcctgacac tcagttccgg gtaggcagtt cgctccaagc tggactgtat
     3301 gcacgaaccc cccgttcagt ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3361 caacccggaa agacatgcaa aagcaccact ggcagcagcc actggtaatt gatttagagg
     3421 agttagtctt gaagtcatgc gccggttaag gctaaactga aaggacaagt tttggtgact
     3481 gcgctcctcc aagccagtta cctcggttca aagagttggt agctcagaga accttcgaaa
     3541 aaccgccctg caaggcggtt ttttcgtttt cagagcaaga gattacgcgc agaccaaaac
     3601 gatctcaaga agatcatctt attaa
//
