LOCUS       sequence-383718-        3220 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses RFP with an engineered CamR-regulated promoter in E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3220)
  AUTHORS   d'Oelsnitz S, Nguyen V, Alper HS, Ellington AD
  TITLE     Evolving a Generalist Biosensor for Bicyclic Monoterpenes.
  JOURNAL   ACS Synth Biol. 2022 Jan 5. doi: 10.1021/acssynbio.1c00402.
  PUBMED    34985281
REFERENCE   2  (bases 1 to 3220)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth 
            Biol. 2022 Jan 5. doi: 10.1021/acssynbio.1c00402."
COMMENT     Sequence Label: Pcamr-RFP
FEATURES             Location/Qualifiers
     source          1..3220
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      429..500
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_RNA        612..661
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     protein_bind    complement(1552..1585)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     promoter        1596..1667
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1668..2483
                     /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             1668..2483
                     /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(1741..1760)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     protein_bind    complement(2617..2650)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     rep_origin      complement(join(2995..3220,1..320))
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(3075..3094)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gctggactgt atgcacgaac cccccgttca gtccgaccgc tgcgccttat ccggtaacta
       61 tcgtcttgag tccaacccgg aaagacatgc aaaagcacca ctggcagcag ccactggtaa
      121 ttgatttaga ggagttagtc ttgaagtcat gcgccggtta aggctaaact gaaaggacaa
      181 gttttggtga ctgcgctcct ccaagccagt tacctcggtt caaagagttg gtagctcaga
      241 gaaccttcga aaaaccgccc tgcaaggcgg ttttttcgtt ttcagagcaa gagattacgc
      301 gcagaccaaa acgatctcaa gaagatcatc ttattaaggg gtctgacgct cagtggaacg
      361 aaaccctgtt ggtcaggttt tcgggaggtg tgcgtctcgc atccggaaag gtgtgatagg
      421 tagcgcagca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt
      481 gtttgtcggt gaacgctctc ctcaacgggc tgtcgacctt tgaaaagttc gtttacagct
      541 agctcagtcc taggtacaat ctcactatat cgccgatata gagcctactg agcacagctg
      601 gctgagcaca gctgtcaccg gatgtgcttt ccggtctgat gagtccgtga ggacgaaaca
      661 gcctctacaa ataattttgt ttaaactaga agtaagcgag gtacacatat ggtcagtaaa
      721 ggagaagctg taattaaaga gtttatgcgc ttcaaagtgc atatggaagg ttccatgaac
      781 ggacatgagt tcgagattga aggtgaaggt gaaggacgcc cctacgaggg cactcaaact
      841 gcaaagttaa aggttacaaa aggaggacct ctgccatttt cgtgggacat tctgagcccg
      901 cagtttatgt acggcagccg cgcgttcatc aaacatcccg ctgacatccc agactattac
      961 aaacaatctt tccccgaagg ctttaaatgg gaacgcgtga tgaactttga agatggtggc
     1021 gctgtgactg tgacccagga cacttcatta gaagatggaa ccctgattta caaggtaaag
     1081 ctgcgcggca ccaactttcc ccctgacgga cctgtaatgc agaaaaaaac aatgggttgg
     1141 gaggctagta cagagcgttt ataccctgag gacggtgtct taaaaggaga catcaagatg
     1201 gcgttacgtc ttaaggatgg tggtcgctat ttagctgact tcaagaccac ttataaagca
     1261 aagaagcccg tccaaatgcc tggagcttat aacgttgacc gtaagttaga catcacctca
     1321 cataacgagg attacacagt tgtcgaacag tatgagcgct cagaaggccg tcattcgact
     1381 ggtggaatgg acgaactgta taaataataa tcctgcttca ttcctcggta ccaaattcca
     1441 gaaaagaggg gagcgggaaa ccgctcccct tttttcgttt tggtccctcg ctgcagtgtc
     1501 gtaaaaacca gttggggaga cgctgtgaat cccgggcaag caaaatcgct ggaagttcct
     1561 atactttcta gagaatagga acttctttct aaatacattc aaatatgtat ccgctcatga
     1621 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agccatattc
     1681 aacgggaaac gtcttgctcc aggccgcgat taaattccaa catggatgct gatttatatg
     1741 ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgattgtatg
     1801 ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg
     1861 ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca
     1921 agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc cccgggaaaa
     1981 cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg
     2041 cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc
     2101 gcgtatttcg cctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg
     2161 attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataagc
     2221 ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta
     2281 tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc
     2341 gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga
     2401 aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt
     2461 tgatgctcga tgagtttttc taagttgtga tggcggtagg aatgtaatcg ttaatccgca
     2521 aataacgtaa aaacccgctt cggcgggttt ttttatgggg ggagtttagg gaaagagcat
     2581 ttgtcatccc gttgaatatg gctcccttaa cgtgaggaag ttcctatact ttctagagaa
     2641 taggaacttc tacagatgga cttgggttgg cggtttcagg agtaggtgct tcctcgctca
     2701 ctgactcgct gcacgaggca gacctcagcg ctagcggagt gtatactggc ttactatgtt
     2761 ggcactgatg agggtgtcag tgaagtgctt catgtggcag gagaaaaaag gctgcaccgg
     2821 tgcgtcagca gaatatgtga tacaggatat attccgcttc ctcgctcact gactcgctac
     2881 gctcggtcgt tcgactgcgg cgagcggaaa tggcttacga acggggcgga gatttcctgg
     2941 aagatgccag gaagatactt aacagggaag tgagagggcc gcggcaaagc cgtttttcca
     3001 taggctccgc ccccctgaca agcatcacga aatctgacgc tcaaatcagt ggtggcgaaa
     3061 cccgacagga ctataaagat accaggcgtt tccccctggc ggctccctcg tgcgctctcc
     3121 tgttcctgcc tttcggttta ccggtgtcat tccgctgtta tggccgcgtt tgtctcattc
     3181 cacgcctgac actcagttcc gggtaggcag ttcgctccaa
//
