LOCUS       sequence-381125-        5722 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses an evolved noscapine biosensor in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5722)
  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 5722)
  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-NOS4
FEATURES             Location/Qualifiers
     source          1..5722
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     63..82
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(102..179)
                     /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        457..491
                     /label=BBa_J23114
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.1"
     misc_RNA        512..561
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             2249..2440
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2434..2456)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2597..2616
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2691..2708)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2862..3450)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2942..2961)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3621..4412,4413..4481))
                     /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"
     promoter        complement(4482..4553)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(join(4741..5722,1..101))
                     /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
                     YIPPLTTIKQDFRLLGQTSVDRSLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
ORIGIN
        1 gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca
       61 ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga
      121 ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcaccattc gatggtgtcg
      181 ggacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt tgcggccgcg
      241 aagacagcgt tatcagagat gagacaccgt gcgataatgt cgggcaatca ggtgcgactc
      301 ggtaccaaat tccagaaaag aggggagcgg gaaaccgctc cccttttttc gttttggtcc
      361 caatctatcg attgtatgga cttcatcttc attacctcgt atcattgtac acctgccgaa
      421 cgcaagggca tgggctgtcg acctttgaaa agtaccttta tggctagctc agtcctaggt
      481 acaatgctag caatcaagat actgagcaca gctgtcaccg gatgtgcttt ccggtctgat
      541 gagtccgtga ggacgaaaca gcctctacaa ataattttgt ttaaactagt gaaccacgag
      601 gcctacatat ggttgctcgc ccaaagtctg aggacaaaaa gcaggcattg cttgaagcgg
      661 caactcaagc catcgcgcaa tcaggcattg ccgctagtac cgctgtaatt gcacgcaatg
      721 cgggagttgc ggaagggacg ttgttccgct atttcgcaac gaaagatgag ttgatcaaca
      781 ccctttactt acatttgacc catgacatgt gccaatcact gatcatggaa ttggatcgtt
      841 ctattactga cgctaagatg atgacccgtt ttatctggaa cagttatatt agctggggat
      901 tgaaccaccc agctcgccat cgtgccattc gtcagttggc ggtttctgaa aagttgacga
      961 aggaaacccg ccaacgcgcc cgcgatatgt tcccggagtt acgcgacttg tgctaccgta
     1021 gtttgcttat ggtgtttatg tccgacgagt accgcgcctt cggcgacggg ttgttcatgg
     1081 cgcttgctga gacgactatg gatttcgctg cgcgcgaccc ggctcgcgct ggtgagtaca
     1141 ttgcgttggg cttcgaagct atgtggcgcg cacttacgcg cgaagagcag taataatcct
     1201 aagaattcga ggagtgcagg ctcggtaaca tacggtctag ctatctgact atcgccgctg
     1261 tgagctcggt accaaattcc agaaaagagg ccgcgaaagc ggcctttttt cgttttggtc
     1321 caagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtatcacta gtcttcgcgg
     1381 ccgccatgct gtccaggcag gtagatgacg accatcaggg acagcttcaa ggatcgctcg
     1441 cggctcttac cagcctaact tcgatcattg gaccgctgat cgtcacggcg atttatgccg
     1501 cctcggcgag cacatggaac gggttggcat ggattgtagg cgccgcccta taccttgtct
     1561 gcctccccgc gttgcgtcgc ggtgcatgga gccgggccac ctcgacctga atggaagccg
     1621 gcggcacctc gctaacggat tcaccactcc aagaattgga gccaatcaat tcttgcggag
     1681 aactgtgaat gcgcaaacca acccttggca gaacatatcc atcgcgtccg ccatctccag
     1741 cagccgcacg cggcgcatct cgggcagcgt tgggtcctgg ccacgggtgc gcatgatcgt
     1801 gctcctgtcg ttgaggaccc ggctaggctg gcggggttgc cttactggtt agcagaatga
     1861 atcaccgata cgcgagcgaa cgtgaagtga ctgctgctgc aaaacgtctg cgacctgagc
     1921 aacaacatga atggtcatcg gtttccgtgt ttcgtaaagt ctggaaacgc ggaagtcagc
     1981 gccctgcacc attatgttcc ggatctgcat cgcaggatgc tgctggctac cctgtggaac
     2041 acctacatct gtattaacga agcgctggca ttgaccctga gtgatttttc tctggtcccg
     2101 ccgcatccat accgccagtt gtttaccctc acaacgttcc agtaaccggg catgttcatc
     2161 atcagtaacc cgtatcgtga gcatcctctc tcgtttcatc ggtatcatta cccccatgaa
     2221 cagaaatccc ccttacacgg aggcatcagt gaccaaacag gaaaaaaccg cccttaacat
     2281 ggcccgcttt atcagaagcc agacattaac gcttctggag aaactcaacg agctggacgc
     2341 ggatgaacag gcagacatct gtgaatcgct tcacgaccac gctgatgagc tttaccgcag
     2401 ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     2461 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     2521 gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta
     2581 tactggctta actatgcggc atcagagcag attgtactga gagtgcaccg gtgtgaaata
     2641 ccgcacagat gcgtaaggag aaaataccgc atcaggcgct cttccgcttc ctcgctcact
     2701 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     2761 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     2821 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2881 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2941 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     3001 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     3061 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     3121 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     3181 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     3241 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     3301 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     3361 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     3421 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     3481 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     3541 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     3601 gagtaaactt ggtctaacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     3661 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     3721 gagggcttac catctggccc cagtgctgca atgataccgc gcgatccacg ctcaccggct
     3781 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     3841 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     3901 ccagttaata gtttgcgcaa cgttgttgcc attgctgcag gcatcgtggt gtcacgctcg
     3961 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     4021 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     4081 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     4141 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     4201 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac gggataatac cgcgccacat
     4261 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     4321 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     4381 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     4441 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     4501 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     4561 aaaaatatgc gccttgagcg acacgaatta tgcagtgatt tacgacctgc acagccatac
     4621 cacagcttcc gatggctgcc tgacgccaga agcattggtg caccgtgcag tcgatgataa
     4681 gctgtcaaac atgagaattg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc
     4741 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     4801 cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga
     4861 gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca gcgagcggtc
     4921 cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata
     4981 acatgagcta tcttcggtat cgtcgtatcc cactaccgag atatccgcac caacgcgcag
     5041 cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat
     5101 cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc
     5161 actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg
     5221 ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat
     5281 ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccat cttcatggga
     5341 gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt
     5401 agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag
     5461 cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct
     5521 tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc
     5581 cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa
     5641 cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat
     5701 cgccgcttcc actttttccc gc
//
