LOCUS       sequence-346383-        5233 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible gene expression, pET origin, CymRAM repressor, PCymRC 
            promoter, mRFP, Carbenicillin Resistance Marker. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5233)
  TITLE     Reisch Lab Duet Plasmids
REFERENCE   2  (bases 1 to 5233)
  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: pECyR4
FEATURES             Location/Qualifiers
     source          1..5233
                     /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"
     rep_origin      126..581
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(213..232)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     423..444
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(696..1487,1488..1556))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRKQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1319..1338
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1557..1649)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1730..2318
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2219..2238
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2472..2489
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2504..2643
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2569..2588)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2729..2751
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2745..2936)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     3508..3524
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4327..4354
                     /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    4385..4412
                     /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        4416..4465
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             4510..4521
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4528..5205
                     /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 tgaaaggagg aactatatcc ggattggcga
      121 atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
      181 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
      241 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
      301 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
      361 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
      421 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
      481 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
      541 atttaacgcg aattttaaca aaatattaac gtttacaatt tctggcggca cgatggcatg
      601 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
      661 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
      721 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
      781 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
      841 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
      901 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttt ccgggaagct
      961 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     1021 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     1081 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     1141 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     1201 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     1261 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     1321 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     1381 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     1441 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     1501 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     1561 ttcctttttc aatcatgatt gaagcattta tcagggttat tgtctcatga gcggatacat
     1621 atttgaatgt atttagaaaa ataaacaaat aggtcatgac caaaatccct taacgtgagt
     1681 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     1741 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     1801 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     1861 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     1921 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     1981 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     2041 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     2101 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     2161 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     2221 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     2281 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     2341 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     2401 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     2461 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc
     2521 tccttacgca tctgtgcggt atttcacacc gcaatggtgc actctcagta caatctgctc
     2581 tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct
     2641 gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     2701 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     2761 tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat
     2821 tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat
     2881 gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat
     2941 gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg
     3001 atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt
     3061 aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag
     3121 cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag
     3181 atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg
     3241 aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt
     3301 cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct
     3361 agccgggtcc tcaacgacag gagcacgatc atgctagtca tgccccgcgc ccaccggaag
     3421 gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg
     3481 agctaactta cattaattgc gttgcgcgta aaacgacggc cagtgatcaa cgtttgaatt
     3541 ttgcataacg ttcacgtgca atttccaggg tgctattacg cacacgttca aaacgttctt
     3601 tatctttctg ccacaggcta cgaactacca gaccacgaac gctgttaaaa atcagccaca
     3661 gaatatcttc ggcatcatca cggctcagac cacggctcac cagcacaccc agccacatat
     3721 cttcaacaac aaaacgatta cgttcaacgg tacgctgaat accttcacgc agtgccggat
     3781 cacgatctgc tgcaacaatc agatccaggc cgatgctaaa atcatcatcc agaaaaaatt
     3841 ctgctgcatc atccagcatc tgctgaataa catcatcttc cggtttcagt tttgccagac
     3901 gtgcacggct acgttcggta atctgctcat acagccattc aaaggttgcc agcagcagtt
     3961 ccagtttggt cggaaaatga tggctctgtg caccacggct aacaccggct gcacccggaa
     4021 catctgcaat acgaaaacct gcataacctt tttcacgcag aacacccagt gctgctgcaa
     4081 tcagtttacc ctgggtttcc attgcacgtt ctgcctgggt acgacgtttc gggctcataa
     4141 gaatctccgt ttggtattta ctatgaatat tccattgggc gctatcatgc cataccgcga
     4201 aaggttttgc gccattcgat ggcgcgccgc cccaattatt gaaggcctcc ctaacggggg
     4261 gccttttttt gtttctggtc tcccgcttaa cgatcgttgg ctgggataca tatgtcaatg
     4321 taccggaaca aacagacaat ctggtctgtt tgtattatgg aaaatttttc tgtataatag
     4381 attcaacaaa cagacaatct ggtctgtttg taaagctgtc accggatgtg ctttccggtc
     4441 tgatgagtcc gtgaggacga aacagcctct acaaataatt ttgtttaaga actgggaaga
     4501 gcgcagatta aagaggagaa atactagatg gcttcctccg aagacgttat caaagagttc
     4561 atgcgtttca aagttcgtat ggaaggttcc gttaacggtc acgagttcga aatcgaaggt
     4621 gaaggtgaag gtcgtccgta cgaaggtacc cagaccgcta aactgaaagt taccaaaggt
     4681 ggtccgctgc cgttcgcttg ggacatcctg tccccgcagt tccagtacgg ttccaaagct
     4741 tacgttaaac acccggctga catcccggac tacctgaaac tgtccttccc ggaaggtttc
     4801 aaatgggaac gtgttatgaa cttcgaagac ggtggtgttg ttaccgttac ccaggactcc
     4861 tccctgcaag acggtgagtt catctacaaa gttaaactgc gtggtaccaa cttcccgtcc
     4921 gacggtccgg ttatgcagaa aaaaaccatg ggttgggaag cttccaccga acgtatgtac
     4981 ccggaagacg gtgctctgaa aggtgaaatc aaaatgcgtc tgaaactgaa agacggtggt
     5041 cactacgacg ctgaagttaa aaccacctac atggctaaaa aaccggttca gctgccgggt
     5101 gcttacaaaa ccgacatcaa actggacatc acctcccaca acgaagacta caccatcgtt
     5161 gaacagtacg aacgtgctga aggtcgtcac tccaccggtg cttaataacg ctgatagtgc
     5221 tagtgtagat cgc
//
