LOCUS       sequence-249114-        5351 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 5351)
  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 5351)
  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: pSPB845
FEATURES             Location/Qualifiers
     source          1..5351
                     /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"""
     protein_bind    185..201
                     /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)."
     protein_bind    208..224
                     /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)."
     CDS             join(322..324,325..327,328..1038)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(367..388)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(628..647)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     975..996
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1039..1095
                     /label=MCS
                     /note="multiple cloning site"
     promoter        2071..2105
                     /label=BBa_J23110
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.33"
     CDS             2142..3224
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(2161..2180)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     misc_feature    3318..3338
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     CDS             complement(3499..4314)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(3499..4314)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4222..4241
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4306..4325
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     terminator      complement(4480..4511)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     primer_bind     complement(4655..4672)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4789..5334)
                     /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(4869..4888)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acggaaaaaa aaaccccgcc cctgacaggg cggggttttt tttccaatta ttgaaggccg
       61 ctaacgcggc ctttttttgt ttctggtctg ccttaatcaa tgactaagaa ttcgcggccg
      121 cttctagagg gtctcacttc ctcgagggcg cgcctgcagg cttcggatcc gctgcaggct
      181 tcaattgtga gcggataaca attgacattg tgagcggata acaagatact gagcacacga
      241 catgcatgcc acttaaaaag gagatcaaca atgaaagcaa ttttcgtact gaaacatctt
      301 aatcatgcta aggagacttc aatggtgagc aagggcgagg agctgttcac cggggtggtg
      361 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
      421 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
      481 ctgcccgtgc cctggcccac cctcgtgacc accctgacct ggggcgtgca gtgcttcagc
      541 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
      601 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
      661 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
      721 gacggcaaca tcctggggca caagctggag tacaactaca tcagccacaa cgtctatatc
      781 accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag
      841 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
      901 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
      961 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     1021 atggacgagc tgtacaagag atctcgagct caagcttcga attctgcagt cgacggtacc
     1081 gcgggcccgg gatccatgtc cgaagggcgt cgtggtctgg gtcgagggct ttcggccctg
     1141 ctgggcgagg tcgacgccgc gccggctcag gcgccgggcg agcagcttgg cggttcccgt
     1201 gaagcgccga tcgaaatcct gcagcgcaat cccgaccagc cccgtcggac cttccgcgag
     1261 gaagacctcg aggatctgtc aaactcgatc cgtgagaagg gcgttctgca gccgatcctg
     1321 gtgcggccgt cgccggatac ggctggcgag taccagatcg tcgccggcga gcgccgttgg
     1381 cgcgcagccc agcgggcggg cctcaagact gtgccgatca tggtgcgcga gctcgacgac
     1441 ctggcggtgc tggagatcgg catcatcgag aacgtccagc gggctgatct gaacgtcctg
     1501 gaagaagcgc tctcctacaa ggtgctgatg gagaagttcg agcgcaccca ggagaacatc
     1561 gcccagacca tcggcaagag ccgcagccac gtggccaaca cgatgcgctt gctggcccta
     1621 ccggacgagg ttcagtcgca cctggtcagt ggtgagctga cggccggcca cgcccgcgcc
     1681 atcgctgcag cggctgaccc ggtggccctc gccaagcaaa tcatcgaggg cggcttgtcg
     1741 gtgcgcgaga cagaggccct ggcccgtaag gcgcccaacc tgtctgcggg caagagcaag
     1801 gggggccgcc ccccacgcgt gaaggacacc gacacccagg cgctggagtc tgacctgtcc
     1861 tcggttctgg gtctggatgt ctcgatcgac catcgcgggt ctaccggcac gctgaccatc
     1921 acctacgcca ccctcgagca gctcgacgac ctctgcaacc gactgacgcg cgggatctaa
     1981 taatggcctc ggtaccaaag acgaacaata agacgctgaa aagcgtcttt tttcgttttg
     2041 gtccgctgcg ttgctagccc tgcaggcttc tttacggcta gctcagtcct aggtacaatg
     2101 ctagctactc gacggaagag agtcaattca gggtggtgaa tgtgaaacca gtaacgttat
     2161 acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg
     2221 ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt
     2281 acattcccaa ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg attggcgttg
     2341 ccacctccag tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg
     2401 ccgatcaact gggtgccagc gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct
     2461 gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc attaactatc
     2521 cgctggatga ccaggatgcc attgctgtgg aagctgcctg cactaatgtt ccggcgttat
     2581 ttcttgatgt ctctgaccag acacccatca acagtattat tttctcccat gaagacggta
     2641 cgcgactggg cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg
     2701 gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa tatctcactc
     2761 gcaatcaaat tcagccgata gcggaacggg aaggcgactg gagtgccatg tccggttttc
     2821 aacaaaccat gcaaatgctg aatgagggca tcgttcccac tgcgatgctg gttgccaacg
     2881 atcagatggc gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg
     2941 acatctcggt agtgggatac gacgataccg aagacagctc atgttatatc ccgccgttaa
     3001 ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac
     3061 tctctcaggg ccaggcggtg aagggcaatc agctgttgcc cgtttcactg gtgaaaagaa
     3121 aaaccaccct ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     3181 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtaataatgg cccaattatt
     3241 gaaggggagc gggaaaccgc tccccttttt ttgtttctgg tctgccgctg gctgccatgg
     3301 cgtttgagac ctgcaggtac tagtagcggc cgctgcagtt aatcactgat taactcggta
     3361 ccaaattcca gaaaagaggc ctcccgaaag gggggccttt tttcgttttg gtccttttgt
     3421 tatcaataaa aaaggggagc ggtttcccgc tccccttatt gttcgtctac agtgttacaa
     3481 ccaattaacc aattctgatt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
     3541 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
     3601 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
     3661 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
     3721 atcaccatga gtgacgactg aatccggtga gaatggcaaa agcttatgca tttctttcca
     3781 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
     3841 gttattcatt cgtgattgcg cctgagcgag gcgaaatacg cgatcgctgt taaaaggaca
     3901 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
     3961 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt
     4021 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
     4081 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
     4141 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt
     4201 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
     4261 gttggaattt aatcgcggcc tggagcaaga cgtttcccgt tgaatatggc tcataacacc
     4321 ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgatgata tatttttatc
     4381 ttgtgcaatg taacatcaga gattttgaga cacaacgtgg ctttgttgaa taaatcgaac
     4441 ttttgctgag ttgaaggatc aggagtcctg ttgagtaata gtcaaaagcc tccggtcgga
     4501 ggcttttgac tttctgctta cccaatagcg gagtgtatac tggcttacta tgttggcact
     4561 gatgagggtg tcagtgaagt gcttcatgtg gcaggagaaa aaaggctgca ccggtgcgtc
     4621 agcagaatat gtgatacagg atatattccg cttcctcgct cactgactcg ctacgctcgg
     4681 tcgttcgact gcggcgagcg gaaatggctt acgaacgggg cggagatttc ctggaagatg
     4741 ccaggaagat acttaacagg gaagtgagag ggccgcggca aagccgtttt tccataggct
     4801 ccgcccccct gacaagcatc acgaaatctg acgctcaaat cagtggtggc gaaacccgac
     4861 aggactataa agataccagg cgtttccccc tggcggctcc ctcgtgcgct ctcctgttcc
     4921 tgcctttcgg tttaccggtg tcattccgct gttatggccg cgtttgtctc attccacgcc
     4981 tgacactcag ttccgggtag gcagttcgct ccaagctgga ctgtatgcac gaaccccccg
     5041 ttcagtccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggaaagac
     5101 atgcaaaagc accactggca gcagccactg gtaattgatt tagaggagtt agtcttgaag
     5161 tcatgcgccg gttaaggcta aactgaaagg acaagttttg gtgactgcgc tcctccaagc
     5221 cagttacctc ggttcaaaga gttggtagct cagagaacct tcgaaaaacc gccctgcaag
     5281 gcggtttttt cgttttcaga gcaagagatt acgcgcagac caaaacgatc tcaagaagat
     5341 catcttatta a
//
