LOCUS       sequence-350294-        3880 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible gene expression, pCOLA origin, VanRAM repressor, PVanCC 
            promoter, mRFP, Kanamycin Resistance Marker. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3880)
  TITLE     Reisch Lab Duet Plasmids
REFERENCE   2  (bases 1 to 3880)
  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: pCVR2
FEATURES             Location/Qualifiers
     source          1..3880
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             61..72
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     misc_RNA        complement(117..166)
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     primer_bind     complement(1136..1153)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1136..1152)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      1350..1985
                     /direction=RIGHT
                     /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     1821..1840
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2003..2094
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2095..2910
                     /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             2095..2910
                     /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     complement(2168..2187)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      complement(3140..3187)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     3183..3201
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(join(3257..3880,1..54))
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
ORIGIN
        1 catacgaact ttgaaacgca tgaactcttt gataacgtct tcggaggaag ccatctagta
       61 tttctcctct ttaatctgcg ctcttcccag ttcttaaaca aaattatttg tagaggctgt
      121 ttcgtcctca cggactcatc agaccggaaa gcacatccgg tgacagctat tggatccaat
      181 ggtacctagg actgagctag ctgtcaattg gatccaatca gccaacgatc gttaagcggg
      241 agaccagaaa caaaaaaagg ccccccgtta gggaggcctt caataattgg cggcgcgcca
      301 tcgaatggcg caaaaccttt cgcggtatgg catgatagcg cccgcttaaa ctaacgaacg
      361 taaataagga ggatagacat ggacatgcct cgtattaaac cgggtcagcg tgttatgatg
      421 gcactgcgta aaatgattgc aagcggtgaa atcaaaagtg gtgaacgtat tgcagaaatt
      481 ccgaccgcag cagcactggg tgttagccgt atgccggttc gtatcgcact gcgttcactg
      541 gaacaagaag gtctggttgt tcgtctgggt gcacgtggtt atgcagcccg tggtgttagc
      601 agcgatcaga ttcgtgatgc aattgaagtt cgtggtgttc tggaaggttt tgcagcacgt
      661 cgtctggcag aacgtggtat gacctcagaa acccatgcac gttttgttgt actgattgca
      721 gaaggtgaag cactgtttgc agccggtcgc ctgaatggtg aagatctgga tcgttatgcc
      781 gcatataatc aggcatttca tgataccctg gttagcgcag caggtaatgg tgcagttgaa
      841 agcgcactgg cacgtaatgg ttttgaaccg tttgcagcag ccggtgcact ggccctggat
      901 ctgatggacc tgtctgccga atatgaacat ctgctggcag cacatcgtca gcatcaggca
      961 gttctggatg cagttagctg tggtgatgcc gaaggtgcag aacgtattat gcgtgatcat
     1021 gcactggcag caattcgtaa tgcaaaagtt tttgaagcag cagcaagcgc aggcgcaccg
     1081 ctgggtgcag catggtcaat tcgtgcagat tgaccatgga ttcttcgtct gtttcactgg
     1141 ccgtcgtttt acacgcaacg caattaatgt aagttagctc actcattagg caccgggatc
     1201 tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat
     1261 gactaacatg agaattacaa cttatatcgt atggggctga cttcaggtgc tacatttgaa
     1321 gagataaatt gcactgaaat ctagagtgat ggtgtcggga atccgtaaag gatcttcttg
     1381 agatcctttt acgatcgtcg taatctcctg ctctgtaaac gaaaaaaccg cctggggagg
     1441 cggtttgatc gaaggttaag tcagttgggg aactgcttaa cctggtaact ggctttagtg
     1501 gagcgcagat accaaatact gtcctttcag tgtagcctct gttaggccac cacttcaaga
     1561 ctctcgatat ctaaatccac taattctcag ttaccaatgg ctgctgccag tggcgttttg
     1621 tcgtgtcttt ccgggttgga ctcaagatga tagttaccgg ataaggcgca gcagtcgggc
     1681 tgaacggggg gttcttgcac acagcccagc ttggagcgaa ctgtctacac ggaacgggac
     1741 gtggtgattt gggtaaagcc tccaccacaa cacggacgcc gcaggacggg aacaggagag
     1801 cgcaagaggg agccatcagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     1861 caccactgat ttgagcgtca gatttcgtga tgttcgtcag gggggcggag cctatggaaa
     1921 aacggcttcg ctccggcctt attgtctctc tgctaagtat cctcctggca tcttctagga
     1981 cgtttctgcg ctagcatgcc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     2041 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagc
     2101 catattcaac gggaaacgtc ttgctctagg ccgcgattaa attccaacat ggatgctgat
     2161 ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcga
     2221 ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc
     2281 aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg
     2341 accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc
     2401 gggaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat
     2461 gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac
     2521 agcgaccgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat
     2581 gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg
     2641 cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat
     2701 aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc
     2761 gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca
     2821 ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag
     2881 tttcatttga tgctcgatga gtttttctaa gaattaattc atgagcggat acatatttga
     2941 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccact
     3001 tgcggagacc cggtcgtcag cttgtcgtcg gttcagggca gggtcgttaa atagccgctt
     3061 atgtctattg ctggtttacc ggtttattga ctaccggaag cagtgtgacc gtgtgcttct
     3121 caaatgcctg aggtttcagc aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt
     3181 atgctagtta ttgctcagcg gtggcagcag gtttcccgac tggaaagcgc gatctacact
     3241 agcactatca gcgttattaa gcaccggtgg agtgacgacc ttcagcacgt tcgtactgtt
     3301 caacgatggt gtagtcttcg ttgtgggagg tgatgtccag tttgatgtcg gttttgtaag
     3361 cacccggcag ctgaaccggt tttttagcca tgtaggtggt tttaacttca gcgtcgtagt
     3421 gaccaccgtc tttcagtttc agacgcattt tgatttcacc tttcagagca ccgtcttccg
     3481 ggtacatacg ttcggtggaa gcttcccaac ccatggtttt tttctgcata accggaccgt
     3541 cggacgggaa gttggtacca cgcagtttaa ctttgtagat gaactcaccg tcttgcaggg
     3601 aggagtcctg ggtaacggta acaacaccac cgtcttcgaa gttcataaca cgttcccatt
     3661 tgaaaccttc cgggaaggac agtttcaggt agtccgggat gtcagccggg tgtttaacgt
     3721 aagctttgga accgtactgg aactgcgggg acaggatgtc ccaagcgaac ggcagcggac
     3781 cacctttggt aactttcagt ttagcggtct gggtaccttc gtacggacga ccttcacctt
     3841 caccttcgat ttcgaactcg tgaccgttaa cggaaccttc
//
