LOCUS       sequence-381132-        6471 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GFP in response to tetrahydropapaverine in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6471)
  AUTHORS   d'Oelsnitz S, Kim W, Burkholder NT, Javanmardi K, Thyer R, Zhang Y, 
            Alper HS, Ellington AD
  TITLE     Using fungible biosensors to evolve improved alkaloid biosyntheses.
  JOURNAL   Nat Chem Biol. 2022 Jul 7. pii: 10.1038/s41589-022-01072-w. doi: 
            10.1038/s41589-022-01072-w.
  PUBMED    35799063
REFERENCE   2  (bases 1 to 6471)
  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: "Nat Chem 
            Biol. 2022 Jul 7. pii: 10.1038/s41589-022-01072-w. doi: 
            10.1038/s41589-022-01072-w."
COMMENT     Sequence Label: pThpR
FEATURES             Location/Qualifiers
     source          1..6471
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             1365..1373
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     primer_bind     complement(2501..2520)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2557..2634
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2635..3717
                     /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"
     rep_origin      3942..4487
                     /direction=RIGHT
                     /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     4388..4407
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4604..4621
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5013..5115
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             5116..5931
                     /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             5116..5931
                     /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(5189..5208)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        6359..6393
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     misc_RNA        6414..6463
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
ORIGIN
        1 aataattttg tttaaactag tgaaccacga ggcctacata tggttgctcg cccaaagtct
       61 gaggacaaaa agcaggcatt gcttgaagcg gcaactcaag ccatcgcgca atcaggcatt
      121 gccgctagta ccgctgtaat tgcacgcaat gcgggagttg cggaagggac gttgttccgc
      181 tatttcgcaa cgaaagatga gttgatcaac accctttact tacatttgtt ccaggactgg
      241 tgccaatcat ccatcatgga attggatcgt tctattactg acgctaagat gatgtcgcgt
      301 tttctctgga acagtatcat tagctgggga ttgaaccacc cagctcgcca tcgtgccata
      361 cgtcagttgg cggtttctga aaagttgacg aaggagaccg aacaacgcgc ggatgatatg
      421 ttcccggagt tacgcgacat cgtccaccgt gaggttctta tggtgtttat gtccgacgag
      481 taccgcgcct tcggcgacgg gttgttcttg gcgcttgctg agacgactat ggatttcgct
      541 gcgcgcaacc cggctcgcgc tggtgagtac attgcgttgg gcttcgaggc tatgtggcgc
      601 gcacttacgc gcgaagagca gtaataatcc taagaattcg aggagtgcag gctcggtaac
      661 atacggtcta gctatctgac tatcgccgct gtgagctcgg taccaaattc cagaaaagag
      721 gccgcgaaag cggccttttt tcgttttggt ccaagcccga tgcgccagag ttgtttctga
      781 aacatggcaa aggtatcaca aggggttatg ctacccgaca ccatcgaatg gccgacagtc
      841 attcatcttt ctgcccctcc aaaagcaaaa acccgccgaa gcgggttttt acgtaaatca
      901 ggtgaaactg accgacgaac taaaacgctt gaggacagcc acacttccaa ctagtctcag
      961 ctaacatata acaggtttgg attattaatc ttgttcttca gcaacaaaat gaacgcaatt
     1021 accagctgga tctctcaaag caaattctct accccaaggt tgttcaccaa tttcggtcat
     1081 agctggacca gaagcatctc taaaattggt agaaacaact tcggaccatt cggcatataa
     1141 ttcatccaaa cctctaaccc aaacccaagc caaagtatta tctggaacaa cttgatcttg
     1201 aacggcggaa atgaacaagg taacatcatc tctaacaaca ccagcgaaat catcttcaac
     1261 gaaatctctg gagaaaccca atctatcagt ccaaaattca acagcaccag caacatctct
     1321 agcggtcaaa actggaacag cagatgtcaa tttagacata tgtaacctcc ttatctaact
     1381 agtttatcac tcgagcttat agagctcatc catgccatga gtaatgcctg ctgcagttac
     1441 aaactccagg agtaccatgt gatcgcgctt ctcatttgga tctttggaga gcacggattg
     1501 agtggagaga taatgattat ctgggaggag tactgggcca tcgccaattg gagtattttg
     1561 ttgataatga tctgcgagtt gtacggagcc atcctccaca ttatggcgaa tcttaaaatt
     1621 cgccttaatg ccattctttt gcttatctgc ggtaatatat acattatggg aattgaaatt
     1681 atactccagc ttatggccga gaatattgcc atcctcctta aaatcaatgc ccttgagctc
     1741 aatgcgattt acgagagtat cgccctcaaa ctttacctct gcgcgagtct tataggtgcc
     1801 atcatcctta aagctaatag tgcgctcttg tacatagccc tctggcattg cggacttaaa
     1861 aaaatcatgt tgcttcatgt gatctggata gcgggaaaaa cattgtacgc cataggtcag
     1921 agtagttacg agagttggcc atggtactgg gagcttgcca gtagtacaaa taaacttgag
     1981 agtgagcttg ccgttagttg catcgccctc gccctcgccg cgtacggaaa acttatggcc
     2041 atttacatcg ccatcgagct ctacgagaat tggtactacg ccagtaaaga gctcctcgcc
     2101 cttggatgcc ataattactc cttattaacc cggaggttta cgttaaacaa aattatttgt
     2161 agaggtcata agtttcgtcg catttcagcg actcatcagt gggcttagcc cagacttatg
     2221 actgactatt gtatgagtga gtaagcactc attatagaaa gcaaagatac gccgtcaagg
     2281 tacttttcaa aggtcgacag cccatgccct tgcgttaact gactagtggt cgccgacgtg
     2341 atagtatgct cgataagatg cgagccatat tcaacgggat gacaaatgct ctttccctaa
     2401 actcccccca taaaaaaacc cgccgaagcg ggtttttacg ttatttgcgg attaacgatt
     2461 acattcctac cgccatcaca acgaaccatt gaggacacag tacaatctgc tctgatgccg
     2521 cggccgcgtc gcataaggga gagcgtcgag atcccggaca ccatcgaatg gcgcaaaacc
     2581 tttcgcggta tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa
     2641 ccagtaacgt tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc
     2701 gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg
     2761 gcggagctga attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg
     2821 ctgattggcg ttgccacctc tagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg
     2881 attaaatctc gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc
     2941 ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg
     3001 atcattaact atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat
     3061 gttccggcgt tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc
     3121 catgaagatg gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc
     3181 gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat
     3241 aaatatctca ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc
     3301 atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg
     3361 ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg
     3421 cgcgttggtg cggatatctc ggtagtggga tacgacgata ccgaagatag ctcatgttat
     3481 atcccgccgt taaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac
     3541 cgcttgctgc aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca
     3601 ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     3661 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     3721 caacgcaatt aatgtaagtt agctcactca ttaggcaccg ggatctcgac cgatgccctt
     3781 gagagccttc aacccagtca gctccttccg gtgggcgcgg ggcatgacta acatgagaat
     3841 tacaacttat atcgtatggg gctgacttca ggtgctacat ttgaagagat aaattgcact
     3901 gaaatctaga aatattttat ctgattaata agatgatctt cttgagatcg ttttggtctg
     3961 cgcgtaatct cttgctctga aaacgaaaaa accgccttgc agggcggttt ttcgaaggtt
     4021 ctctgagcta ccaactcttt gaaccgaggt aactggcttg gaggagcgca gtcaccaaaa
     4081 cttgtccttt cagtttagcc ttaaccggcg catgacttca agactaactc ctctaaatca
     4141 attaccagtg gctgctgcca gtggtgcttt tgcatgtctt tccgggttgg actcaagacg
     4201 atagttaccg gataaggcgc agcggtcgga ctgaacgggg ggttcgtgca tacagtccag
     4261 cttggagcga actgcctgcc cggaactgag tgtcaggcgt ggaatgagac aaacgcggcc
     4321 ataacagcgg aatgacaccg gtaaaccgaa aggcaggaac aggagagcgc acgagggagc
     4381 cgccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac cactgatttg
     4441 agcgtcagat ttcgtgatgc ttgtcagggg ggcggagcct atggaaaaac ggctttgccg
     4501 cggccatctc acttccctgt taagtatctt cctggcatct tccaggaaat ctccgccccg
     4561 ttcgtaagcc atttccgctc gccgcagtcg aacgaccgag cgtagcgagt cagtgagcga
     4621 ggaagcggaa tatatcctgt atcacatatt ctgctgacgc accggtgcag ccttttttct
     4681 cctgccacat gaagcacttc actgacaccc tcatcagtgc caacatagta agccagtata
     4741 cactccgcta gcgctgatgt ccggcggtgc ttttgccgtt acgcaccacc ccgtcagtag
     4801 ctgaacagga gggacagctg atagaaacag aagccactgg agcacctcaa aaacaccatc
     4861 atacactaaa tcagtaagtt ggcagcatca cccgacgcac tttgcgccga ataaatacct
     4921 gtgacggaag atcacttcgc agaataaata aatcctggtg tccctgttga taccgggaag
     4981 ccctgggcca acttttggcg aaaatgagac gttgatcggc acgtaagagg ttccaacttt
     5041 caccataatg aaataagatc actaccgggc gtattttttg agttatcgag attttcagga
     5101 gctaaggaag ctaaaatgag ccatattcaa cgggaaacgt cttgctctag gccgcgatta
     5161 aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa
     5221 tcaggtgcga caatctatcg attgtatggg aagcccgatg cgccagagtt gtttctgaaa
     5281 catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg
     5341 acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg
     5401 ttactcacca ctgcgatccc cgggaaaaca gcattccagg tattagaaga atatcctgat
     5461 tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct
     5521 gtttgtaatt gtccttttaa cagcgaccgc gtatttcgtc tcgctcaggc gcaatcacga
     5581 atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt
     5641 gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact
     5701 catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt
     5761 gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc
     5821 ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat
     5881 cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta attttttaag
     5941 gcagttattg gtgcccttaa acgcctggtt gctacgcctg aataagtgat aataagcgga
     6001 tgaatggcag aaattcgaaa gcaaattcga cccggtcgtc ggttcagggc agggtcgtta
     6061 aatagccgct tatgtctatt gctggtttac cggtttattg actaccggaa gcagtgtgag
     6121 cggccgcgcc gagttctaac gacaatatgc gttatcagag atgagacacc gtgcgataat
     6181 gtcgggcaat caggtgcgac tcggtaccaa attccagaaa agaggggagc gggaaaccgc
     6241 tccccttttt tcgttttggt cccaatctat cgattgtatg gacttcatct tcattacctc
     6301 gtatcattgt acacctgccg aacgcaaggg catgggctgt cgacctttga aaagtacctt
     6361 tacggctagc tcagtcctag gtatagtgct agcaatcaag atactgagca cagctgtcac
     6421 cggatgtgct ttccggtctg atgagtccgt gaggacgaaa cagcctctac a
//
