LOCUS       sequence-383670-        5698 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the camphor-responsive biosensor CamR in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5698)
  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 5698)
  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"; journalName: "ACS Synth 
            Biol. 2022 Jan 5. doi: 10.1021/acssynbio.1c00402."
COMMENT     Sequence Label: pReg-CamR
FEATURES             Location/Qualifiers
     source          1..5698
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        23..72
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     regulatory      448..457
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1736..1927
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1921..1943)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2084..2103
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2178..2195)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2349..2937)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2429..2448)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3108..3899,3900..3968))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPXAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3731..3750
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3969..4040)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(4228..5310)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     5272..5291
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5311..5388)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     promoter        join(5666..5698,1..2)
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
ORIGIN
        1 gcaatcaaga tactgagcac agctgtcacc ggatgtgctt tccggtctga tgagtccgtg
       61 aggacgaaac agcctctaca aataattttg tttaaactag ctgccgcaga cccgcacata
      121 tggacattaa gcagtcgtta cttcacgcag ctatgcgctt acttagcgcc aagggccgcg
      181 acggagctac catgcgtgca atctgcgccg aagtgggtgt tactcctcca actctgtatc
      241 accattatgg cgatcttcaa ggtttacata aagcagctat cgatgaaaca taccgtcagg
      301 tcgcagaggc atatcacggc ggaaccgagg agcgtggacc gttaaaaggc atccgcgacg
      361 gctgggccac attcctgcag ttcgcctatt cagagccaaa tatgtgccgt atgttggttc
      421 aacatatcat ggcaggagaa cccccttcca tggtggctga tacccttcgc ggagtcgcag
      481 atgatttagc tcagttccat gcacaaggcc gcttaacctt cccaccgcgt gaggccgccc
      541 aattattatg gatgggcgct ttgggtgcac tgacctatgc actttcacgt gaaggagcag
      601 gttacactca ggacttggca ctgcagaagg ctaaattaga tattacactt gtagcactgt
      661 ttaatatcga ggaggaataa taatcctaag aattcgagga gtgcaggctc ggtaacatac
      721 ggtctagcta tctgactatc gccgctgtga gctcggtacc aaattccaga aaagaggccg
      781 cgaaagcggc cttttttcgt tttggtccaa gcccgatgcg ccagagttgt ttctgaaaca
      841 tggcaaaggt atcactagtc ttcgcggccg ccatgctgtc caggcaggta gatgacgacc
      901 atcagggaca gcttcaagga tcgctcgcgg ctcttaccag cctaacttcg atcattggac
      961 cgctgatcgt cacggcgatt tatgccgcct cggcgagcac atggaacggg ttggcatgga
     1021 ttgtaggcgc cgccctatac cttgtctgcc tccccgcgtt gcgtcgcggt gcatggagcc
     1081 gggccacctc gacctgaatg gaagccggcg gcacctcgct aacggattca ccactccaag
     1141 aattggagcc aatcaattct tgcggagaac tgtgaatgcg caaaccaacc cttggcagaa
     1201 catatccatc gcgtccgcca tctccagcag ccgcacgcgg cgcatctcgg gcagcgttgg
     1261 gtcctggcca cgggtgcgca tgatcgtgct cctgtcgttg aggacccggc taggctggcg
     1321 gggttgcctt actggttagc agaatgaatc accgatacgc gagcgaacgt gaagtgactg
     1381 ctgctgcaaa acgtctgcga cctgagcaac aacatgaatg gtcatcggtt tccgtgtttc
     1441 gtaaagtctg gaaacgcgga agtcagcgcc ctgcaccatt atgttccgga tctgcatcgc
     1501 aggatgctgc tggctaccct gtggaacacc tacatctgta ttaacgaagc gctggcattg
     1561 accctgagtg atttttctct ggtcccgccg catccatacc gccagttgtt taccctcaca
     1621 acgttccagt aaccgggcat gttcatcatc agtaacccgt atcgtgagca tcctctctcg
     1681 tttcatcggt atcattaccc ccatgaacag aaatccccct tacacggagg catcagtgac
     1741 caaacaggaa aaaaccgccc ttaacatggc ccgctttatc agaagccaga cattaacgct
     1801 tctggagaaa ctcaacgagc tggacgcgga tgaacaggca gacatctgtg aatcgcttca
     1861 cgaccacgct gatgagcttt accgcagctg cctcgcgcgt ttcggtgatg acggtgaaaa
     1921 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     1981 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac
     2041 ccagtcacgt agcgatagcg gagtgtatac tggcttaact atgcggcatc agagcagatt
     2101 gtactgagag tgcaccggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     2161 aggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     2221 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     2281 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     2341 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     2401 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     2461 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     2521 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     2581 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     2641 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     2701 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     2761 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     2821 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     2881 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     2941 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     3001 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     3061 agttttaaat caatctaaag tatatatgag taaacttggt ctaacagtta ccaatgctta
     3121 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     3181 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     3241 ataccgcgcg atccacgctc accggctcca gatttatcag caataaacca gccagccgga
     3301 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     3361 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     3421 gctrcaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     3481 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     3541 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     3601 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     3661 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     3721 tcaacacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     3781 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     3841 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     3901 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     3961 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     4021 agcggataca tatttgaatg tatttagaaa aatatgcgcc ttgagcgaca cgaattatgc
     4081 agtgatttac gacctgcaca gccataccac agcttccgat ggctgcctga cgccagaagc
     4141 attggtgcac cgtgcagtcg atgataagct gtcaaacatg agaattgtgc ctaatgagtg
     4201 agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     4261 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     4321 cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg
     4381 gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg
     4441 tttgatggtg gttaacggcg ggatataaca tgagctatct tcggtatcgt cgtatcccac
     4501 taccgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag
     4561 cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg
     4621 catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg
     4681 aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga
     4741 acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac
     4801 gcccagtcgc gtaccatctt catgggagaa aataatactg ttgatgggtg tctggtcaga
     4861 gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg
     4921 gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac
     4981 cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc
     5041 cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag
     5101 actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg
     5161 gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga
     5221 aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc
     5281 tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg
     5341 gcgctatcat gccataccgc gaaaggtttt gcaccattcg atggtgtcgg gacgtcaggt
     5401 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt gcggccgcga agacagcgtt
     5461 atcagagatg agacaccgtg cgataatgtc gggcaatcag gtgcgactcg gtaccaaatt
     5521 ccagaaaaga ggggagcggg aaaccgctcc ccttttttcg ttttggtccc aatctatcga
     5581 ttgtatggac ttcatcttca ttacctcgta tcattgtaca cctgccgaac gcaagggcat
     5641 gggctgtcga cctttgaaaa gtacctttac ggctagctca gtcctaggta tagtgcta
//
