LOCUS       sequence-346572-        3802 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 3802)
  TITLE     Reisch Lab Duet Plasmids
REFERENCE   2  (bases 1 to 3802)
  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..3802
                     /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"
     protein_bind    2896..2923
                     /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    2954..2981
                     /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        2985..3034
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             3079..3090
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3097..3774
                     /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 agtgatcaac
     2101 gtttgaattt tgcataacgt tcacgtgcaa tttccagggt gctattacgc acacgttcaa
     2161 aacgttcttt atctttctgc cacaggctac gaactaccag accacgaacg ctgttaaaaa
     2221 tcagccacag aatatcttcg gcatcatcac ggctcagacc acggctcacc agcacaccca
     2281 gccacatatc ttcaacaaca aaacgattac gttcaacggt acgctgaata ccttcacgca
     2341 gtgccggatc acgatctgct gcaacaatca gatccaggcc gatgctaaaa tcatcatcca
     2401 gaaaaaattc tgctgcatca tccagcatct gctgaataac atcatcttcc ggtttcagtt
     2461 ttgccagacg tgcacggcta cgttcggtaa tctgctcata cagccattca aaggttgcca
     2521 gcagcagttc cagtttggtc ggaaaatgat ggctctgtgc accacggcta acaccggctg
     2581 cacccggaac atctgcaata cgaaaacctg cataaccttt ttcacgcaga acacccagtg
     2641 ctgctgcaat cagtttaccc tgggtttcca ttgcacgttc tgcctgggta cgacgtttcg
     2701 ggctcataag aatctccgtt tggtatttac tatgaatatt ccattgggcg ctatcatgcc
     2761 ataccgcgaa aggttttgcg ccattcgatg gcgcgccgcc ccaattattg aaggcctccc
     2821 taacgggggg cctttttttg tttctggtct cccgcttaac gatcgttggc tgggatacat
     2881 atgtcaatgt accggaacaa acagacaatc tggtctgttt gtattatgga aaatttttct
     2941 gtataataga ttcaacaaac agacaatctg gtctgtttgt aaagctgtca ccggatgtgc
     3001 tttccggtct gatgagtccg tgaggacgaa acagcctcta caaataattt tgtttaagaa
     3061 ctgggaagag cgcagattaa agaggagaaa tactagatgg cttcctccga agacgttatc
     3121 aaagagttca tgcgtttcaa agttcgtatg gaaggttccg ttaacggtca cgagttcgaa
     3181 atcgaaggtg aaggtgaagg tcgtccgtac gaaggtaccc agaccgctaa actgaaagtt
     3241 accaaaggtg gtccgctgcc gttcgcttgg gacatcctgt ccccgcagtt ccagtacggt
     3301 tccaaagctt acgttaaaca cccggctgac atcccggact acctgaaact gtccttcccg
     3361 gaaggtttca aatgggaacg tgttatgaac ttcgaagacg gtggtgttgt taccgttacc
     3421 caggactcct ccctgcaaga cggtgagttc atctacaaag ttaaactgcg tggtaccaac
     3481 ttcccgtccg acggtccggt tatgcagaaa aaaaccatgg gttgggaagc ttccaccgaa
     3541 cgtatgtacc cggaagacgg tgctctgaaa ggtgaaatca aaatgcgtct gaaactgaaa
     3601 gacggtggtc actacgacgc tgaagttaaa accacctaca tggctaaaaa accggttcag
     3661 ctgccgggtg cttacaaaac cgacatcaaa ctggacatca cctcccacaa cgaagactac
     3721 accatcgttg aacagtacga acgtgctgaa ggtcgtcact ccaccggtgc ttaataacgc
     3781 tgatagtgct agtgtagatc gc
//
