LOCUS       sequence-493816-        6405 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of PopZ fused to mCherry and sfGFP under the control of 
            the PopZ promoter (KanR) .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6405)
  AUTHORS   Lu N, Duvall SW, Zhao G, Kowallis KA, Zhang C, Tan W, Sun J, 
            Petitjean HN, Tomares DT, Zhao GP, Childers WS, Zhao W
  TITLE     Scaffold-Scaffold Interaction Facilitates Cell Polarity Development 
            in Caulobacter crescentus.
  JOURNAL   mBio. 2023 Apr 25;14(2):e0321822. doi: 10.1128/mbio.03218-22. Epub 
            2023 Mar 27.
  PUBMED    36971555
REFERENCE   2  (bases 1 to 6405)
  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"; journalName: "mBio."; 
            date: "2023-04-25"; volume: "14(2):e0321822. doi"; pages: " 
            10.1128/mbio.03218-22. Epub 2023 Mar 27"
COMMENT     Sequence Label: pLN053
FEATURES             Location/Qualifiers
     source          1..6405
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            21..130
                     /note="incP origin of transfer"
     primer_bind     282..298
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      299..345
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             2156..2863
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2301..2319)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2569..2588
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2879..3592
                     /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
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(4229..4245)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4256..4273)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4256..4272)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4265..4287)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(4470..5058)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4550..4569)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5468..6283)
                     /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(5468..6283)
                     /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     6191..6210
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6275..6294
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 ccaccgcggt ggcggccgcc ccggccagcc tcgcagagca ggattcccgt tgagcaccgc
       61 caggtgcgaa taagggacag tgaagaagga acacccgctc gcgggtgggc ctacttcacc
      121 tatcctgccc ggctgacgcc gttggataca ccaaggaaag tctacacgaa ccctttggca
      181 aaatcctgta tatcgtgcga aaaaggatgg atataccgaa aaaatcgcta taatgacccc
      241 gaagcagggt tatgcagcgg aaaagatccg tcgacggccg ctctagaact agtggatcca
      301 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcgca
      361 ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt caggttcatc atgccgtttg
      421 tgatggcttc catgtcggca gaatgcttaa tgaattacaa cagtttttac cgtcgcaacg
      481 tttgcgtgaa aaaagcttaa ctggcgacga ctggcggagc gacttacacg cgatcatatt
      541 gtggccggcg cgagcttcgc gtgattgggg atagaatgtt gatgtcgaac cgtcgacggg
      601 ccgggttgct ggccgccgct tgcagcgctg gtcttctggc cgggtccttc tccatcgcga
      661 acgccgagac tctcgccgat gcgatcgcgc tcgcctatca gaccaatccg acgctgcagc
      721 aacagcgcgc cagcgcccgc atcaccgacg aaggcgtggt gcaagccaag accggcttcc
      781 gcccgaccgt gggcgccacc gccgatatca cgggggcgaa caccaatccg gccagcgccg
      841 gcgcggatgt gaagctctct ggcagcagcg cggcgctttc gatcatccag ccgctctaca
      901 ccggcggtcg cgccagcgcg ggcgtcagcg ccgctgaagc cgacgtgctg tctgcgcggg
      961 aaggtcttcg cgcggtcgag cagggggtgc tggtcagcgt cgtccaggcc tatgtcgacg
     1021 tgcgccgaga ccaggaacgc ctgcgcatcg ccaaggaaaa cgtcgcggtt ctgcagcgcc
     1081 agctcgaaga atcgaacgct cgcttcgacg tgggtgagat cacccggacg gacgtcgccc
     1141 agtctcaggc gcgcttggct tcggccaagg ccagcctgtc gggcgcccag gcccagttgg
     1201 aagtcagccg cgcctcctac gctgcggtgg tcggtcaaac gcccggcgaa ctggctcccg
     1261 agccgagctt ggccggactg ctgcccgcca gcgtcgacca ggctttcgag gccgcccaga
     1321 cgaacaatcc tggcgtgacc tcggcgaagt tcaatgaaga ggcggccgcc gcgcgtgtcg
     1381 ccgtcgccaa ggcgggctat cgaccgacgg tctcggcgcg cgcttcgctg ggttacgatg
     1441 cgacgcgtcg ttctggcgtc ggcagccagt ttgacgacta ttcgcgcacc gtttcgggcg
     1501 cgatcacggc ctcgatcccg cttttcgccg gcggcttgac cacgtcgcag gtccgggcgg
     1561 ccaatgaacg tgagagcgct gcgcgcagcg ccgtcgaagg cgccaagcgc accgcgatcc
     1621 agcaagtctc gaacgcgtgg agcaatctgc tggcgtcgcg cgccggtctg gtcgccaacg
     1681 aggagcaggt tcgagcgacc cgcatcgctt tcgaaggcgt gcgtcaggag caacaggtcg
     1741 gcctgcggac gacgctggac gttctgaacg cccagctgga gctgtccaac gccgaactgg
     1801 cgctggtttc ggcccgtcgc gacgaatacc tggcttccgc cgtggtcttg caggccatgg
     1861 gccaactgga ggtgaccaag ctcgcctcca acaccgccgc ctatgacccg aaaacctcct
     1921 acaaccgcgt gacgcgcggc gtgggctggg tgccgtggga gccggtggtg caggcgatcg
     1981 acagcgtcgg cgcggcttcc accaagcccc cgagcggctc gaaataggtt tgatccgcgt
     2041 cgattccgcc caggcgcgga cttgcgcttg cttaacacgc gtaagtctgc accatcgcgg
     2101 ctaagcgcgc cccaaggcgc gcaaggattc gtacctcttt acgacggccc cgcacatggt
     2161 gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca
     2221 catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc
     2281 ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc
     2341 ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc
     2401 cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat
     2461 gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga
     2521 gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca
     2581 gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct
     2641 gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt
     2701 caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat
     2761 caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc
     2821 cgagggccgc cactccaccg gcggcatgga cgagctgtac aagcaccggt cggccaccat
     2881 gcgtaaaggc gaagagctgt tcactggtgt cgtccctatt ctggtggaac tggatggtga
     2941 tgtcaacggt cataagtttt ccgtgcgtgg cgagggtgaa ggtgacgcaa ctaatggtaa
     3001 actgacgctg aagttcatct gtactactgg taaactgccg gtaccttggc cgactctggt
     3061 aacgacgctg acttatggtg ttcagtgctt tgctcgttat ccggaccata tgaagcagca
     3121 tgacttcttc aagtccgcca tgccggaagg ctatgtgcag gaacgcacga tttcctttaa
     3181 ggatgacggc acgtacaaaa cgcgtgcgga agtgaaattt gaaggcgata ccctggtaaa
     3241 ccgcattgag ctgaaaggca ttgactttaa agaagacggc aatatcctgg gccataagct
     3301 ggaatacaat tttaacagcc acaatgttta catcaccgcc gataaacaaa aaaatggcat
     3361 taaagcgaat tttaaaattc gccacaacgt ggaggatggc agcgtgcagc tggctgatca
     3421 ctaccagcaa aacactccaa tcggtgatgg tcctgttctg ctgccagaca atcactatct
     3481 gagcacgcaa agcgttctgt ctaaagatcc gaacgagaaa cgcgatcata tggttctgct
     3541 ggagttcgta accgcagcgg gcatcacgca tggtatggat gaactgtaca aaggtggcag
     3601 tggtagcatg tccgatcagt ctcaagaacc tacaatggag gaaatcctcg cctccattcg
     3661 acgcatcatc tcggaggatg acgcgccggc ggagcctgcg gccgaagcgg cgcccccgcc
     3721 gccgccggaa cccgaacctg aaccggtgtc gttcgacgac gaggttctgg aattgacgga
     3781 tccgatcgcg cccgagcccg agctgccgcc gctggagact gtcggcgaca tcgacgtcta
     3841 ttcgccgccg gaacctgagt cggaaccggc ctacacgccg ccgccggcgg ctccggtgtt
     3901 tgatcgcgac gaagtcgccg agcagctggt cggcgtttcg gccgcttcgg ccgcggcgag
     3961 cgccttcggc agcctgagct cggccctgct gatgcccaag gacggtcgga cgctggaaga
     4021 cgtcgtacgc gagctgctgc gcccgctgct caaggagtgg ctggaccaga acctgccgcg
     4081 catcgtcgag accaaggttg aggaagaagt gcagcgtatc tctcggggac gcggcgccta
     4141 agccttaatt aatatgcatg gtaccttaag atctcgagct ccggagaatt cgaacgttac
     4201 gcgtaccggt gctagctgca gcccggggga tccactagtt ctagagcggc cattcactgg
     4261 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     4321 cagcacatcc ccctttcgcc agcggtatca gctcactcaa aggcggtaat acggttatcc
     4381 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4441 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4501 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     4561 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4621 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     4681 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4741 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4801 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4861 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     4921 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     4981 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     5041 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5101 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     5161 atccttttag gggggggggg gcgctgaggt ctgcctcgtg aagaaggtgt tgctgactca
     5221 taccaggcct gaatcgcccc atcatccagc cagaaagtga gggagccacg gttgatgaga
     5281 gctttgttgt aggtggacca gttggtgatt ttgaactttt gctttgccac ggaacggtct
     5341 gcgttgtcgg gaagatgcgt gatctgatcc ttcaactcag caaaagttcg atttattcaa
     5401 caaagccgcc gtcccgtcaa gtcagcgtaa tgctctgcca gtgttacaac caattaacca
     5461 attctgatta gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat
     5521 tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc
     5581 agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa
     5641 tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag
     5701 tgacgactga atccggtgag aatggcaaaa gcttatgcat ttctttccag acttgttcaa
     5761 caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc
     5821 gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag
     5881 gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt tcacctgaat
     5941 caggatattc ttctaatacc tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc
     6001 atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca
     6061 gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt
     6121 tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc gcacctgatt
     6181 gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta
     6241 atcgcggcct cgagcaagac gtttcccgtt gaatatggct cataacaccc cttgtattac
     6301 tgtttatgta agcagacagt tttattgttc atgatgatat atttttatct tgtgcaatgt
     6361 aacatcagag attttgagac acaacgtggc tttccccccc ccccc
//
