LOCUS       sequence-270548-        4603 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CmR, Para-popZ-rbs-H3H4-msfGFPN-CTM. Replicative plasmid for the 
            expression of bait. N-terminal GFP fusion..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4603)
  AUTHORS   Lim HC, Bernhardt TG
  TITLE     A PopZ-Linked Apical Recruitment Assay for Studying Protein-Protein 
            Interactions in the Bacterial Cell Envelope.
  JOURNAL   Mol Microbiol. 2019 Sep 24. doi: 10.1111/mmi.14391.
  PUBMED    31550057
REFERENCE   2  (bases 1 to 4603)
  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: "Mol 
            Microbiol. 2019 Sep 24. doi: 10.1111/mmi.14391."
COMMENT     Sequence Label: pHCL149
FEATURES             Location/Qualifiers
     source          1..4603
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..285
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             2341..2532
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2526..2548)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2634..2774
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2689..2708
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2789..2806)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2960..3548)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3040..3059)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3719..4378)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4293..4312
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4379..4481)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcga
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg tttttttggg
      301 tctagcagga gatacatata tgtccgatca gtctcaagaa cctacaatgg aggaaatcct
      361 cgcctccatt cgacgcatca tctcggagga tgacgcgccg gcggagcctg cggccgaagc
      421 ggcgcccccg ccgccgccgg aacccgaacc tgaaccggtg tcgttcgacg acgaggttct
      481 ggaattgacg gacccgatcg cgcccgagcc cgagctgccg ccgctggaga ctgtcggcga
      541 catcgacgtc tattcgccgc cggaacctga gtcggaaccg gcctacacgc cgccgccggc
      601 ggctccggtg tttgatcgcg acgaagtcgc cgagcagctg gtcggcgttt cggccgcttc
      661 ggccgcggcg agcgccttcg gcagcctgag ctcggccctg ctgatgccca aggacggtcg
      721 gacgctggaa gacgtcgtac gcgagctgct gcgcccgctg ctcaaggagt ggctggacca
      781 gaacctgccg cgcatcgtcg agaccaaggt tgaggaagaa gtgcagcgta tctctcgggg
      841 acgcggctaa cgctgtctag aaggagatat acatatgagt gcgttattaa tgcctaagga
      901 tggccgcaca cttgaagatg ttgttagaga acttctgcgt cctttgctta aagaatggct
      961 tgatcaaaat ttgcctcgca ttgttgagac gaaagtggaa gaagaggttc aacgtatttc
     1021 gcggggtaga ggcggaggtg gttctaaggg agaagagttg tttacgggtg ttgtaccgat
     1081 tcttgtcgaa ctggacggag atgtgaatgg acataaattt tcggtccgtg gcgagggaga
     1141 gggtgacgct accaacggaa aactcactct taagttcatc tgtaccaccg gcaagctgcc
     1201 tgtcccatgg ccaacccttg tgacgactct gacttatggc gtgcaatgtt tttcgcggta
     1261 tcccgaccac atgaaacagc atgatttttt caagtcggcg atgccagaag gatacgtgca
     1321 agagcggacg atctccttca aggatgacgg cacctacaag acccgtgccg aagttaagtt
     1381 tgagggcgat actttggtca atcggattga gctgaaagga atcgatttca aagaagacgg
     1441 caatatcctg ggacacaagc tcgaatataa ttttaacagc cacaatgtat atattactgc
     1501 ggataaacag aagaatggca tcaaggcaaa ctttaaaatt cgccataacg tagaagatgg
     1561 atcggtccaa ctcgccgacc actaccagca gaacactcca atcggagatg gcccggtttt
     1621 gcttccagat aaccattatc tcagcactca atcaaagctc agcaaagacc ccaacgagaa
     1681 gcgggatcac atggtactgt tggagttcgt aaccgctgcc ggtatcacgc acggtatgga
     1741 cgaattgtat aaaggttctg gcggtggatc cacgcgtccc cgatcccccc gttcccgtaa
     1801 cagcaaagca cgcccggcac ccggcctgaa caaatggctc agctgggctc tcaagctcgg
     1861 cctcgtcggc ctggtgctcc tcgccggctt cgccatctat ctcgatgccg tggtgcagga
     1921 gaaattctcc ggcctcgagg tcgactaagc ttatcaccga tacgcgagcg aacgtgaagc
     1981 gactgctgct gcaaaacgtc tgcgacctga gcaacaacat gaatggtctt cggtttccgt
     2041 gtttcgtaaa gtctggaaac gcggaagtca gcgccctgca ccattatgtt ccggatctgc
     2101 atcgcaggat gctgctggct accctgtgga acacctacat ctgtattaac gaagcgctgg
     2161 cattgaccct gagtgatttt tctctggtcc cgccgcatcc ataccgccag ttgtttaccc
     2221 tcacaacgtt ccagtaaccg ggcatgttca tcatcagtaa cccgtatcgt gagcatcctc
     2281 tctcgtttca tcggtatcat tacccccatg aacagaaatc ccccttacac ggaggcatca
     2341 gtgaccaaac aggaaaaaac cgcccttaac atggcccgct ttatcagaag ccagacatta
     2401 acgcttctgg agaaactcaa cgagctggac gcggatgaac aggcagacat ctgtgaatcg
     2461 cttcacgacc acgctgatga gctttaccgc agctgcctcg cgcgtttcgg tgatgacggt
     2521 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     2581 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc
     2641 atgacccagt cacgtagcga tagcggagtg tatactggct taactatgcg gcatcagagc
     2701 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     2761 aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     2821 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     2881 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     2941 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3001 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3061 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3121 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     3181 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3241 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3301 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3361 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     3421 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3481 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3541 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3601 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     3661 attaaaaatg aagttttaaa tcaatctaaa gcggccgcct tcatttaaat ggcgcgcctt
     3721 acgccccgcc ctgccactca tcgcagtact gttgtaattc attaagcatt ctgccgacat
     3781 ggaagccatc acaaacggca tgatgaacct gaatcgccag cggcatcagc accttgtcgc
     3841 cttgcgtata atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc atattggcca
     3901 cgtttaaatc aaaactggtg aaactcaccc agggattggc tgagacgaaa aacatattct
     3961 caataaaccc tttagggaaa taggccaggt tttcaccgta acacgccaca tcttgcgaat
     4021 atatgtgtag aaactgccgg aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt
     4081 cagtttgctc atggaaaacg gtgtaacaag ggtgaacact atcccatatc accagctcac
     4141 cgtctttcat tgccatacgt aattccggat gagcattcat caggcgggca agaatgtgaa
     4201 taaaggccgg ataaaacttg tgcttatttt tctttacggt ctttaaaaag gccgtaatat
     4261 ccagctgaac ggtctggtta taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt
     4321 ctttacgatg ccattgggat atatcaacgg tggtatatcc agtgattttt ttctccattt
     4381 tagcttcctt agctcctgaa aatctcgaca actcaaaaaa tacgcccggt agtgatctta
     4441 tttcattatg gtgaaagttg gaacctctta cgtgccgatc aacgtctcat tttcgccaaa
     4501 agttggccca gggcttcccg gtatcaacag ggacaccagg atttatttat tctgcgaagt
     4561 gatcttccgt cacaggtaga gatctgaatt ccgccattca gag
//
