LOCUS       sequence-251668-        7086 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  asymmetric plasmid partitioning E. coli - target plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7086)
  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 7086)
  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: pEE29
FEATURES             Location/Qualifiers
     source          1..7086
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..474
                     /gene="sopC"
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
     CDS             597..3671
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-galactosidase"
                     /label=lacZ
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYC
                     QDSRLPSEFDLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQ
                     ISDFHVATRFNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIID
                     ERGGYADRVTLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIEN
                     GLLLLNGKPLLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLW
                     YTLCDRYGLYVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWS
                     LGNESGHGANHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVP
                     KWSIKKWLSLPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSL
                     IKYDENGNPWSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQ
                     TIEVTSEYLFRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQ
                     LWLTVRVVQPNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELG
                     NKRWQFNRQSGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWK
                     AAGHYQAEAALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDV
                     EVASDTPHPARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPY
                     VFPSENGLRCGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDG
                     FHMGIGGDDSWSPSVSAEFQLSAGRYHYQLVWCQK"
     primer_bind     complement(617..634)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(617..633)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(626..648)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             3837..4550
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFDGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             4740..4784
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(4822..4840)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4836..4883
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(5054..5845)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASTGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTSKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(5846..5937)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5985..6723)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     primer_bind     complement(6937..6958)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ggcctgaata ttctctctgg gcccactgtt ccacttgtat cgtcggtctg ataatcagac
       61 tgggaccacg gtcccactcg tatcgtcggt ctgattatta gtctgggacc acggtcccac
      121 tcgtatcgtc ggtctgatta ttagtctggg accacggtcc cactcgtatc gtcggtctga
      181 taatcagact gggaccacgg tcccactcgt atcgtcggtc tgattattag tctgggacca
      241 tggtcccact cgtatcgtcg gtctgattat tagtctggga ccacggtccc actcgtatcg
      301 tcggtctgat tattagtctg gaaccacggt cccactcgta tcgtcggtct gattattagt
      361 ctgggaccac ggtcccactc gtatcgtcgg tctgattatt agtctgggac cacgatccca
      421 ctcgtgttgt cggtctgatt atcggtctgg gaccacggtc ccacttgtat tgtcgatcag
      481 actatcagcg tgagactacg attccatcaa tgcctgtcaa gggcaagtat tgacatgtcg
      541 tcgtaacctg tagaacggag taacctcggt gcaagaagac aaaaatcacc ttcaacatga
      601 ccatgattac ggattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
      661 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagtgaag
      721 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgctttg
      781 cctggtttcc ggcaccagaa gcggtgccgg aaagctggct ggagtgcgat cttcctgagg
      841 ccgatactgt cgtcgtcccc tcaaactggc agatgcacgg ttacgatgcg cccatctaca
      901 ccaacgtgac ctatcccatt acggtcaatc cgccgtttgt tcccacggag aatccgacgg
      961 gttgttactc gctcacattt aatgttgatg aaagctggct acaggaaggc cagacgcgaa
     1021 ttatttttga tggcgttaac tcggcgtttc atctgtggtg caacgggcgc tgggtcggtt
     1081 actgccagga cagtcgtttg ccgtctgaat ttgacctgag cgcattttta cgcgccggag
     1141 aaaaccgcct cgcggtgatg gtgctgcgct ggagtgacgg cagttatctg gaagatcagg
     1201 atatgtggcg gatgagcggc attttccgtg acgtctcgtt gctgcataaa ccgactacac
     1261 aaatcagcga tttccatgtt gccactcgct ttaatgatga tttcagccgc gctgtactgg
     1321 aggctgaagt tcagatgtgc ggcgagttgc gtgactacct acgggtaaca gtttctttat
     1381 ggcagggtga aacgcaggtc gccagcggca ccgcgccttt cggcggtgaa attatcgatg
     1441 agcgtggtgg ttatgccgat cgcgtcacac tacgtctgaa cgtcgaaaac ccgaaactgt
     1501 ggagcgccga aatcccgaat ctctatcgtg cggtggttga actgcacacc gccgacggca
     1561 cgctgattga agcagaagcc tgcgatgtcg gtttccgcga ggtgcggatt gaaaatggtc
     1621 tgctgctgct gaacggcaag ccgttgctga ttcgaggcgt taaccgtcac gagcatcatc
     1681 ctctgcatgg tcaggtcatg gatgagcaga cgatggtgca ggatatcctg ctgatgaagc
     1741 agaacaactt taacgccgtg cgctgttcgc attatccgaa ccatccgctg tggtacacgc
     1801 tgtgcgaccg ctacggcctg tatgtggtgg atgaagccaa tattgaaacc cacggcatgg
     1861 tgccaatgaa tcgtctgacc gatgatccgc gctggctacc ggcgatgagc gaacgcgtaa
     1921 cgcgaatggt gcagcgcgat cgtaatcacc cgagtgtgat catctggtcg ctggggaatg
     1981 aatcaggcca cggcgctaat cacgacgcgc tgtatcgctg gatcaaatct gtcgatcctt
     2041 cccgcccggt gcagtatgaa ggcggcggag ccgacaccac ggccaccgat attatttgcc
     2101 cgatgtacgc gcgcgtggat gaagaccagc ccttcccggc tgtgccgaaa tggtccatca
     2161 aaaaatggct ttcgctacct ggagagacgc gcccgctgat cctttgcgaa tacgcccacg
     2221 cgatgggtaa cagtcttggc ggtttcgcta aatactggca ggcgtttcgt cagtatcccc
     2281 gtttacaggg cggcttcgtc tgggactggg tggatcagtc gctgattaaa tatgatgaaa
     2341 acggcaaccc gtggtcggct tacggcggtg attttggcga tacgccgaac gatcgccagt
     2401 tctgtatgaa cggtctggtc tttgccgacc gcacgccgca tccagcgctg acggaagcaa
     2461 aacaccagca gcagtttttc cagttccgtt tatccgggca aaccatcgaa gtgaccagcg
     2521 aatacctgtt ccgtcatagc gataacgagc tcctgcactg gatggtggcg ctggatggta
     2581 agccgctggc aagcggtgaa gtgcctctgg atgtcgctcc acaaggtaaa cagttgattg
     2641 aactgcctga actaccgcag ccggagagcg ccgggcaact ctggctcaca gtacgcgtag
     2701 tgcaaccgaa cgcgaccgca tggtcagaag ccgggcacat cagcgcctgg cagcagtggc
     2761 gtctggcgga aaacctcagt gtgacgctcc ccgccgcgtc ccacgccatc ccgcatctga
     2821 ccaccagcga aatggatttt tgcatcgagc tgggtaataa gcgttggcaa tttaaccgcc
     2881 agtcaggctt tctttcacag atgtggattg gcgataaaaa acaactgctg acgccgctgc
     2941 gcgatcagtt cacccgtgca ccgctggata acgacattgg cgtaagtgaa gcgacccgca
     3001 ttgaccctaa cgcctgggtc gaacgctgga aggcggcggg ccattaccag gccgaagcag
     3061 cgttgttgca gtgcacggca gatacacttg ctgatgcggt gctgattacg accgctcacg
     3121 cgtggcagca tcaggggaaa accttattta tcagccggaa aacctaccgg attgatggta
     3181 gtggtcaaat ggcgattacc gttgatgttg aagtggcgag cgatacaccg catccggcgc
     3241 ggattggcct gaactgccag ctggcgcagg tagcagagcg ggtaaactgg ctcggattag
     3301 ggccgcaaga aaactatccc gaccgcctta ctgccgcctg ttttgaccgc tgggatctgc
     3361 cattgtcaga catgtatacc ccgtacgtct tcccgagcga aaacggtctg cgctgcggga
     3421 cgcgcgaatt gaattatggc ccacaccagt ggcgcggcga cttccagttc aacatcagcc
     3481 gctacagtca acagcaactg atggaaacca gccatcgcca tctgctgcac gcggaagaag
     3541 gcacatggct gaatatcgac ggtttccata tggggattgg tggcgacgac tcctggagcc
     3601 cgtcagtatc ggcggaattc cagctgagcg ccggtcgcta ccattaccag ttggtctggt
     3661 gtcaaaaata agaagacaaa aatcaccttg cgctaagagc tcgacaccat cgaatggtgc
     3721 aaaacctttc gcggtatggc atgatagcgc ccggaagcat gccacttaaa aaggagatca
     3781 acaatgaaag caattttcgt actgaaacat cttaatcatg ctggggaggg tttctaatgc
     3841 gtaaaggcga agagctgttc actggtgtcg tccctattct ggtggaactg gatggtgatg
     3901 tcaacggtca taagttttcc gtgcgtggcg agggtgaagg tgacgcaact aatggtaaac
     3961 tgacgctgaa gttcatctgt actactggta aactgccggt accttggccg actctggtaa
     4021 cgacgctgac ttatggtgtt cagtgctttg ctcgttatcc ggaccatatg aagcagcatg
     4081 acttcttcaa gtccgccatg ccggaaggct atgtgcagga acgcacgatt tcctttaagg
     4141 atgacggcac gtacaaaacg cgtgcggaag tgaaatttga cggcgatacc ctggtaaacc
     4201 gcattgagct gaaaggcatt gactttaaag aagacggcaa tatcctgggc cataagctgg
     4261 aatacaattt taacagccac aatgtttaca tcaccgccga taaacaaaaa aatggcatta
     4321 aagcgaattt taaaattcgc cacaacgtgg aggatggcag cgtgcagctg gctgatcact
     4381 accagcaaaa cactccaatc ggtgatggtc ctgttctgct gccagacaat cactatctga
     4441 gctaccaaag cgttctgtct aaagatccga acgagaaacg cgatcatatg gttctgctgg
     4501 agttcgtaac cgcagcgggc atcacgcatg gtatggatga actgtacaaa acctccgcag
     4561 cgaacgacga aaattacgtg cgccatcacg ccgcgcatct cgcgctccga cagcacgcgc
     4621 aggatgctgc tggatcttag tatattagtt aagtataaga aggagatata catatggcag
     4681 atctcaattg gatatcggcc ggccacgcga tcgctgacgt cggtaccctc gagtctggta
     4741 aagaaaccgc tgctgcgaaa tttgaacgcc agcacatgga ctcgtctact agcgcagctt
     4801 aattaaccta ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta
     4861 aacgggtctt gaggggtttt ttgctgaaac ctcaggcatt tgagaagcac acggtcacac
     4921 tgcttccggt agtcaataaa ccggtaaacc agcaatagac ataagcggct atttaacgac
     4981 cctgccctga accgacgacc gggtcatcgt ggccggatct tgcggcccct cggcttgaac
     5041 gaattgttag acattatttg ccgactacct tggtgatctc gcctttcacg tagtggacaa
     5101 attcttccaa ctgatctgcg cgcgaggcca agcgatcttc ttcttgtcca agataagcct
     5161 gtctagcttc aagtatgacg ggctgatact gggccggcag gcgctccatt gcccagtcgg
     5221 cagcgacatc cttcggcgcg attttgctgg ttactgcgct gtaccaaatg cgggacaacg
     5281 taagcactac atttcgctca tcgccagccc agtcgggcgg cgagttccat agcgttaagg
     5341 tttcatttag cgcctcaaat agatcctgtt caggaaccgg atcaaagagt tcctccgccg
     5401 ctggacctac caaggcaacg ctatgttctc ttgcttttgt cagcaagata gccagatcaa
     5461 tgtcgatcgt ggctggctcg aagatacctg caagaatgtc attgcgctgc cattctccaa
     5521 attgcagttc gcgcttagct ggataacgcc acggaatgat gtcgtcgtgc acaacaatgg
     5581 tgacttctac agcgcggaga atctcgctct ctccagtgga agccgaagtt tccaaaaggt
     5641 cgttgatcaa agctcgccgc gttgtttcat caagccttac ggtcaccgta accagcaaat
     5701 caatatcact gtgtggcttc aggccgccat ccactgcgga gccgtacaaa tgtacggcca
     5761 gcaacgtcgg ttcgagatgg cgctcgatga cgccaactac ctctgatagt tgagtcgata
     5821 cttcggcgat caccgcttcc ctcatactct tcctttttca atattattga agcatttatc
     5881 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     5941 ctagctcact cggtcgctac gctccgggcg tgagactgcg gcgggcgctg cggacacata
     6001 caaagttacc cacagattcc gtggataagc aggggactaa catgtgaggc aaaacagcag
     6061 ggccgcgccg gtggcgtttt tccataggct ccgccctcct gccagagttc acataaacag
     6121 acgcttttcc ggtgcatctg tgggagccgt gaggctcaac catgaatctg acagtacggg
     6181 cgaaacccga caggacttaa agatccccac cgtttccggc gggtcgctcc ctcttgcgct
     6241 ctcctgttcc gaccctgccg tttaccggat acctgttccg cctttctccc ttacgggaag
     6301 tgtggcgctt tctcatagct cacacactgg tatctcggct cggtgtaggt cgttcgctcc
     6361 aagctgggct gtaagcaaga actccccgtt cagcccgact gctgcgcctt atccggtaac
     6421 tgttcacttg agtccaaccc ggaaaagcac ggtaaaacgc cactggcagc agccattggt
     6481 aactgggagt tcgcagagga tttgtttagc taaacacgcg gttgctcttg aagtgtgcgc
     6541 caaagtccgg ctacactgga aggacagatt tggttgctgt gctctgcgaa agccagttac
     6601 cacggttaag cagttcccca actgacttaa ccttcgatca aaccacctcc ccaggtggtt
     6661 ttttcgttta cagggcaaaa gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     6721 atcttttcta ctgaaccgct ctagatttca gtgcaattta tctcttcaaa tgtagcacct
     6781 gaagtcagcc ccatacgata taagttgtaa ttctcatgtt agtcatgccc cgcgcccacc
     6841 ggaaggagct gactgggttg aaggctctca agggcatcgg tcgagatccc ggtgcctaat
     6901 gagtgagcta acttacatta attgcgttgc gccacaccag tttggaacaa gagtccacct
     6961 cgagctttca ggaaaatttt tctgtataat agattcataa atttgagaga agagtgcgac
     7021 cacgttttag tctacgttta tctgtcttta cttaatgtcc tttgttacag gccagaaagc
     7081 ataact
//
