LOCUS       sequence-334893-        3831 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible gene expression, pCOLA origin, CymRAM repressor, PCymRC 
            promoter, mRFP, Kanamycin Resistance Marker. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3831)
  TITLE     Reisch Lab Duet Plasmids
REFERENCE   2  (bases 1 to 3831)
  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"
COMMENT     Sequence Label: pCCyR2
FEATURES             Location/Qualifiers
     source          1..3831
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(28..46)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      42..89
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(319..1134)
                     /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(319..1134)
                     /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     1042..1061
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1135..1226)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1244..1879)
                     /direction=LEFT
                     /label=ColA ori
                     /note="Plasmids containing the ColA origin of replication 
                     can be propagated in E. coli cells that contain additional 
                     plasmids with compatible origins."
     primer_bind     complement(1389..1408)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2077..2093
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2106..2122
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2925..2952
                     /label=CuO
                     /bound_moiety="CymR"
                     /note="CymR-binding P2 operator sequence from the p-cmt 
                     operon of Pseudomonas putida (Mullick et al., 2006)"
     protein_bind    2983..3010
                     /label=CuO
                     /bound_moiety="CymR"
                     /note="CymR-binding P2 operator sequence from the p-cmt 
                     operon of Pseudomonas putida (Mullick et al., 2006)"
     misc_RNA        3014..3063
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             3108..3119
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3126..3803
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
ORIGIN
        1 gctttccagt cgggaaacct gctgccaccg ctgagcaata actagcataa ccccttgggg
       61 cctctaaacg ggtcttgagg ggttttttgc tgaaacctca ggcatttgag aagcacacgg
      121 tcacactgct tccggtagtc aataaaccgg taaaccagca atagacataa gcggctattt
      181 aacgaccctg ccctgaaccg acgacaagct gacgaccggg tctccgcaag tggcactttt
      241 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
      301 ccgctcatga attaattctt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
      361 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
      421 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
      481 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
      541 atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca
      601 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
      661 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cggtcgctgt taaaaggaca
      721 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
      781 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt
      841 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
      901 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
      961 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt
     1021 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
     1081 gttggaattt aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc tcatactctt
     1141 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     1201 tgaatgtatt tagaaaaata aacaaatagg catgctagcg cagaaacgtc ctagaagatg
     1261 ccaggaggat acttagcaga gagacaataa ggccggagcg aagccgtttt tccataggct
     1321 ccgcccccct gacgaacatc acgaaatctg acgctcaaat cagtggtggc gaaacccgac
     1381 aggactataa agataccagg cgtttccccc tgatggctcc ctcttgcgct ctcctgttcc
     1441 cgtcctgcgg cgtccgtgtt gtggtggagg ctttacccaa atcaccacgt cccgttccgt
     1501 gtagacagtt cgctccaagc tgggctgtgt gcaagaaccc cccgttcagc ccgactgctg
     1561 cgccttatcc ggtaactatc atcttgagtc caacccggaa agacacgaca aaacgccact
     1621 ggcagcagcc attggtaact gagaattagt ggatttagat atcgagagtc ttgaagtggt
     1681 ggcctaacag aggctacact gaaaggacag tatttggtat ctgcgctcca ctaaagccag
     1741 ttaccaggtt aagcagttcc ccaactgact taaccttcga tcaaaccgcc tccccaggcg
     1801 gttttttcgt ttacagagca ggagattacg acgatcgtaa aaggatctca agaagatcct
     1861 ttacggattc ccgacaccat cactctagat ttcagtgcaa tttatctctt caaatgtagc
     1921 acctgaagtc agccccatac gatataagtt gtaattctca tgttagtcat gccccgcgcc
     1981 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc
     2041 taatgagtga gctaacttac attaattgcg ttgcgcgtaa aacgacggcc agtggtcacg
     2101 acgttgtaaa acgacggcca gtgatcaacg tttgaatttt gcataacgtt cacgtgcaat
     2161 ttccagggtg ctattacgca cacgttcaaa acgttcttta tctttctgcc acaggctacg
     2221 aactaccaga ccacgaacgc tgttaaaaat cagccacaga atatcttcgg catcatcacg
     2281 gctcagacca cggctcacca gcacacccag ccacatatct tcaacaacaa aacgattacg
     2341 ttcaacggta cgctgaatac cttcacgcag tgccggatca cgatctgctg caacaatcag
     2401 atccaggccg atgctaaaat catcatccag aaaaaattct gctgcatcat ccagcatctg
     2461 ctgaataaca tcatcttccg gtttcagttt tgccagacgt gcacggctac gttcggtaat
     2521 ctgctcatac agccattcaa aggttgccag cagcagttcc agtttggtcg gaaaatgatg
     2581 gctctgtgca ccacggctaa caccggctgc acccggaaca tctgcaatac gaaaacctgc
     2641 ataacctttt tcacgcagaa cacccagtgc tgctgcaatc agtttaccct gggtttccat
     2701 tgcacgttct gcctgggtac gacgtttcgg gctcataaga atctccgttt ggtatttact
     2761 atgaatattc cattgggcgc tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg
     2821 cgcgccgccc caattattga aggcctccct aacggggggc ctttttttgt ttctggtctc
     2881 ccgcttaacg atcgttggct gggatacata tgtcaatgta ccggaacaaa cagacaatct
     2941 ggtctgtttg tattatggaa aatttttctg tataatagat tcaacaaaca gacaatctgg
     3001 tctgtttgta aagctgtcac cggatgtgct ttccggtctg atgagtccgt gaggacgaaa
     3061 cagcctctac aaataatttt gtttaagaac tgggaagagc gcagattaaa gaggagaaat
     3121 actagatggc ttcctccgaa gacgttatca aagagttcat gcgtttcaaa gttcgtatgg
     3181 aaggttccgt taacggtcac gagttcgaaa tcgaaggtga aggtgaaggt cgtccgtacg
     3241 aaggtaccca gaccgctaaa ctgaaagtta ccaaaggtgg tccgctgccg ttcgcttggg
     3301 acatcctgtc cccgcagttc cagtacggtt ccaaagctta cgttaaacac ccggctgaca
     3361 tcccggacta cctgaaactg tccttcccgg aaggtttcaa atgggaacgt gttatgaact
     3421 tcgaagacgg tggtgttgtt accgttaccc aggactcctc cctgcaagac ggtgagttca
     3481 tctacaaagt taaactgcgt ggtaccaact tcccgtccga cggtccggtt atgcagaaaa
     3541 aaaccatggg ttgggaagct tccaccgaac gtatgtaccc ggaagacggt gctctgaaag
     3601 gtgaaatcaa aatgcgtctg aaactgaaag acggtggtca ctacgacgct gaagttaaaa
     3661 ccacctacat ggctaaaaaa ccggttcagc tgccgggtgc ttacaaaacc gacatcaaac
     3721 tggacatcac ctcccacaac gaagactaca ccatcgttga acagtacgaa cgtgctgaag
     3781 gtcgtcactc caccggtgct taataacgct gatagtgcta gtgtagatcg c
//
