LOCUS       sequence-381127-        5720 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses an evolved papaverine biosensor in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5720)
  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 5720)
  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: pReg-PAP4
FEATURES             Location/Qualifiers
     source          1..5720
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      925..1513
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1414..1433
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1667..1684
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(1759..1778)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1919..1941
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1935..2126)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        complement(3882..3916)
                     /label=BBa_J23112
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0"
     promoter        complement(3882..3916)
                     /label=BBa_J23103
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.01"
     promoter        4194..4271
                     /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."
     CDS             4272..5354
                     /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     complement(4291..4310)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        5542..5613
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5614..5682,5683..5720,1..754)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
       61 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
      121 cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc
      181 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
      241 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
      301 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
      361 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
      421 tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact
      481 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
      541 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
      601 tgagcgtgga tcgcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
      661 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
      721 tgagataggt gcctcactga ttaagcattg gtaactgtta gaccaagttt actcatatat
      781 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
      841 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
      901 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
      961 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     1021 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     1081 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     1141 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     1201 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     1261 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     1321 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     1381 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     1441 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     1501 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     1561 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     1621 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     1681 cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     1741 acaccggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact
     1801 ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac
     1861 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     1921 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg
     1981 taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc
     2041 agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta
     2101 agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg
     2161 ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa
     2221 catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac
     2281 cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca
     2341 cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac
     2401 ttccgcgttt ccagacttta cgaaacacgg aaaccgatga ccattcatgt tgttgctcag
     2461 gtcgcagacg ttttgcagca gcagtcactt cacgttcgct cgcgtatcgg tgattcattc
     2521 tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc
     2581 atgcgcaccc gtggccagga cccaacgctg cccgagatgc gccgcgtgcg gctgctggag
     2641 atggcggacg cgatggatat gttctgccaa gggttggttt gcgcattcac agttctccgc
     2701 aagaattgat tggctccaat tcttggagtg gtgaatccgt tagcgaggtg ccgccggctt
     2761 ccattcaggt cgaggtggcc cggctccatg caccgcgacg caacgcgggg aggcagacaa
     2821 ggtatagggc ggcgcctaca atccatgcca acccgttcca tgtgctcgcc gaggcggcat
     2881 aaatcgccgt gacgatcagc ggtccaatga tcgaagttag gctggtaaga gccgcgagcg
     2941 atccttgaag ctgtccctga tggtcgtcat ctacctgcct ggacagcatg gcggccgcga
     3001 agactagtga tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttggaccaa
     3061 aacgaaaaaa ggccgctttc gcggcctctt ttctggaatt tggtaccgag ctcacagcgg
     3121 cgatagtcag atagctagac cgtatgttac cgagcctgca ctcctcgaat tcttaggatt
     3181 attactgctc ttcgcgcgta agtgcgcgcc acatagcctc gaagcccaac gcaatgtact
     3241 caccagcgcg agccgggtcg cgcgcagcga aatccatagt cgtctcagca agcgccaaga
     3301 acaacccgtc gccgaaggcg cggtactcgt cggacataaa caccataaga accctacggt
     3361 gcaggatgtc gcgtaactcc gggaacatat ccaggtcgcg ttggtgggtt tccttcgtca
     3421 acttttcaga aaccgccaac tgacgaatgg cacgatggcg agctgggtgg ttcaatcccc
     3481 agctaatgcc actgttccag ataaaacgca tcatcatctt agcgtcagta atagaacgat
     3541 ccaattccat gatgagtgat tggcacaggt cctgcctcaa atgtaagtaa agggtgttga
     3601 tcaactcatc tttcgttgcg aaatagcgga acaacgtccc ttccgcaact cccgcattgc
     3661 gtgcaattac agcggtacta gcggcaatgc ctgattgcgc gatggcttga gttgccgctt
     3721 caagcaatgc ctgctttttg tcctcagact ttgggcgagc aaccatatac taacctcctt
     3781 ctgatacgtg gttccgttaa acaaaattat ttgtagaggc cccatttcgt ccttttggac
     3841 tcatcagggg tggtacacac caccctatgg ggctcgtaat tgctagcata atccctagga
     3901 ctgagctagc tatcagggta cttttcaaag gtcgacagcc catgcccttg cgttcggcag
     3961 gtgtacaatg atacgaggta atgaagatga agtccataca atcgatagat tgggaccaaa
     4021 acgaaaaaag gggagcggtt tcccgctccc ctcttttctg gaatttggta ccgagtcgca
     4081 cctgattgcc cgacattatc gcacggtgtc tcatctctga taacgctgtc ttcgcggccg
     4141 caacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cccgacacca
     4201 tcgaatggtg caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca
     4261 gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt
     4321 atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa
     4381 aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg
     4441 cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt
     4501 cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt
     4561 cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc
     4621 aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg
     4681 aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca
     4741 acagtattat tttctcccat gaagatggta cgcgactggg cgtggagcat ctggtcgcat
     4801 tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc
     4861 gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata gcggaacggg
     4921 aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg aatgagggca
     4981 tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca atgcgcgcca
     5041 ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt agtgggatac gacgataccg
     5101 aagatagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg
     5161 ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc
     5221 agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg
     5281 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     5341 aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc tcactcatta ggcacaattc
     5401 tcatgtttga cagcttatca tcgactgcac ggtgcaccaa tgcttctggc gtcaggcagc
     5461 catcggaagc tgtggtatgg ctgtgcaggt cgtaaatcac tgcataattc gtgtcgctca
     5521 aggcgcatat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     5581 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     5641 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     5701 aaagtaaaag atgctgaaga
//
