LOCUS       sequence-251662-        4192 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 4192)
  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 4192)
  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: pSPB867
FEATURES             Location/Qualifiers
     source          1..4192
                     /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             join(168..170,171..173,174..884)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(213..234)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(474..493)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     821..842
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    885..941
                     /label=MCS
                     /note="multiple cloning site"
     misc_feature    1906..1926
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     CDS             complement(join(2154..2945,2946..3014))
                     /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     2777..2796
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3015..3119)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(3163..3194)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     primer_bind     complement(3338..3355)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3472..4017)
                     /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(3552..3571)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             4084..4095
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_feature    4142..4163
                     /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 gtgagcaagg
      181 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
      241 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
      301 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
      361 tgacctgggg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
      421 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
      481 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
      541 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
      601 actacatcag ccacaacgtc tatatcaccg ccgacaagca gaagaacggc atcaaggcca
      661 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
      721 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
      781 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
      841 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagagatct cgagctcaag
      901 cttcgaattc tgcagtcgac ggtaccgcgg gcccgggatc catgtccgaa gggcgtcgtg
      961 gtctgggtcg agggctttcg gccctgctgg gcgaggtcga cgccgcgccg gctcaggcgc
     1021 cgggcgagca gcttggcggt tcccgtgaag cgccgatcga aatcctgcag cgcaatcccg
     1081 accagccccg tcggaccttc cgcgaggaag acctcgagga tctgtcaaac tcgatccgtg
     1141 agaagggcgt tctgcagccg atcctggtgc ggccgtcgcc ggatacggct ggcgagtacc
     1201 agatcgtcgc cggcgagcgc cgttggcgcg cagcccagcg ggcgggcctc aagactgtgc
     1261 cgatcatggt gcgcgagctc gacgacctgg cggtgctgga gatcggcatc atcgagaacg
     1321 tccagcgggc tgatctgaac gtcctggaag aagcgctctc ctacaaggtg ctgatggaga
     1381 agttcgagcg cacccaggag aacatcgccc agaccatcgg caagagccgc agccacgtgg
     1441 ccaacacgat gcgcttgctg gccctaccgg acgaggttca gtcgcacctg gtcagtggtg
     1501 agctgacggc cggccacgcc cgcgccatcg ctgcagcggc tgacccggtg gccctcgcca
     1561 agcaaatcat cgagggcggc ttgtcggtgc gcgagacaga ggccctggcc cgtaaggcgc
     1621 ccaacctgtc tgcgggcaag agcaaggggg gccgcccccc acgcgtgaag gacaccgaca
     1681 cccaggcgct ggagtctgac ctgtcctcgg ttctgggtct ggatgtctcg atcgaccatc
     1741 gcgggtctac cggcacgctg accatcacct acgccaccct cgagcagctc gacgacctct
     1801 gcaaccgact gacgcgcggg atctaataat ggcctcggta ccaaagacga acaataagac
     1861 gctgaaaagc gtcttttttc gttttggtcc gctgcgtttg agacgtacta gtagcggccg
     1921 ctgcagttaa tcactgatta actcggtacc aaattccaga aaagaggcct cccgaaaggg
     1981 gggccttttt tcgttttggt ccttttgtta tcaataaaaa aggggagcgg tttcccgctc
     2041 cccttattgt tcgtctacag attatcaaaa aggatcttca cctagatcct tttaaattaa
     2101 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     2161 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     2221 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     2281 gcaatgatac cgcgggaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     2341 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     2401 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     2461 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     2521 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     2581 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     2641 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     2701 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     2761 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     2821 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     2881 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     2941 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     3001 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     3061 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     3121 acatttcccc gaaaagtgcc acctggagtc ctgttgagta atagtcaaaa gcctccggtc
     3181 ggaggctttt gactttctgc ttacccaata gcggagtgta tactggctta ctatgttggc
     3241 actgatgagg gtgtcagtga agtgcttcat gtggcaggag aaaaaaggct gcaccggtgc
     3301 gtcagcagaa tatgtgatac aggatatatt ccgcttcctc gctcactgac tcgctacgct
     3361 cggtcgttcg actgcggcga gcggaaatgg cttacgaacg gggcggagat ttcctggaag
     3421 atgccaggaa gatacttaac agggaagtga gagggccgcg gcaaagccgt ttttccatag
     3481 gctccgcccc cctgacaagc atcacgaaat ctgacgctca aatcagtggt ggcgaaaccc
     3541 gacaggacta taaagatacc aggcgtttcc ccctggcggc tccctcgtgc gctctcctgt
     3601 tcctgccttt cggtttaccg gtgtcattcc gctgttatgg ccgcgtttgt ctcattccac
     3661 gcctgacact cagttccggg taggcagttc gctccaagct ggactgtatg cacgaacccc
     3721 ccgttcagtc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggaaa
     3781 gacatgcaaa agcaccactg gcagcagcca ctggtaattg atttagagga gttagtcttg
     3841 aagtcatgcg ccggttaagg ctaaactgaa aggacaagtt ttggtgactg cgctcctcca
     3901 agccagttac ctcggttcaa agagttggta gctcagagaa ccttcgaaaa accgccctgc
     3961 aaggcggttt tttcgttttc agagcaagag attacgcgca gaccaaaacg atctcaagaa
     4021 gatcatctta ttaaacggaa aaaaaaaccc cgcccctgac agggcggggt tttttttcca
     4081 attattgaag gccgctaacg cggccttttt ttgtttctgg tctgccttaa tcaatgacta
     4141 agaattcgcg gccgcttcta gagcgtctca cttcggatcc gctgcaggct tc
//
