LOCUS       sequence-173109-        5915 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mWasabi reporter gene driven by GrpE promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5915)
  AUTHORS   Piraner DI, Abedi MH, Moser BA, Lee-Gosselin A, Shapiro MG
  TITLE     Tunable thermal bioswitches for in vivo control of microbial 
            therapeutics.
  JOURNAL   Nat Chem Biol. 2017 Jan;13(1):75-80. doi: 10.1038/nchembio.2233. 
            Epub 2016 Nov 14.
  PUBMED    27842069
REFERENCE   2  (bases 1 to 5915)
  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"; doi: 
            "10.1038/nchembio.2233"; journalName: "Nat Chem Biol"; date: 
            "2017-01"; volume: "13"; issue: "1"; pages: "75-80"
COMMENT     Sequence Label: pGrpE-Wasabi
FEATURES             Location/Qualifiers
     source          1..5915
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..714
                     /codon_start=1
                     /product="green variant of teal fluorescent protein mTFP1 
                     (Ai et al., 2008)"
                     /label=mWasabi
                     /translation="MVSKGEETTMGVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDG
                     TNTINLEVKEGAPLPFSYDILTTAFSYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTF
                     EDKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKETTGWDASTERMYVRDGVLK
                     GDVKMKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYEIA
                     VARNSTDGMDELYKG"
     CDS             717..761
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(799..817)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      813..860
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      897..1352
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(984..1003)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1194..1215
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(1467..2258,2259..2327))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2090..2109
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2328..2420)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2501..3089
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2990..3009
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3243..3260
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3275..3414
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3340..3359)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3500..3522
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3516..3707)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4279..5361)
                     /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     5323..5342
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5362..5439)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5645..5664
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             5886..5903
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 atggtgagca agggcgagga gaccacaatg ggcgtaatca agcccgacat gaagatcaag
       61 ctgaagatgg agggcaacgt gaatggccac gccttcgtga tcgagggcga gggcgagggc
      121 aagccctacg acggcaccaa caccatcaac ctggaggtga aggagggagc ccccctgccc
      181 ttctcctacg acattctgac caccgcgttc agttacggca acagggcctt caccaagtac
      241 cccgacgaca tccccaacta cttcaagcag tccttccccg agggctactc ttgggagcgc
      301 accatgacct tcgaggacaa gggcatcgtg aaggtgaagt ccgacatctc catggaggag
      361 gactccttca tctacgagat acacctcaag ggcgagaact tcccccccaa cggccccgtg
      421 atgcagaagg agaccaccgg ctgggacgcc tccaccgaga ggatgtacgt gcgcgacggc
      481 gtgctgaagg gcgacgtcaa gatgaagctg ctgctggagg gcggcggcca ccaccgcgtt
      541 gacttcaaga ccatctacag ggccaagaag gcggtgaagc tgcccgacta tcactttgtg
      601 gaccaccgca tcgagatcct gaaccacgac aaggactaca acaaggtgac cgtttacgag
      661 atcgccgtgg cccgcaactc caccgacggc atggacgagc tgtacaaggg ctgagtaaag
      721 aaaccgctgc tgcgaaattt gaacgccagc acatggactc gtctactagc gcagcttaat
      781 taacctaggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac
      841 gggtcttgag gggttttttg ctgaaaggag gaactatatc cggattggcg aatgggacgc
      901 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
      961 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     1021 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     1081 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     1141 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     1201 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     1261 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     1321 gaattttaac aaaatattaa cgtttacaat ttctggcggc acgatggcat gagattatca
     1381 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     1441 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     1501 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     1561 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     1621 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     1681 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     1741 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     1801 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     1861 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     1921 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     1981 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     2041 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     2101 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     2161 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     2221 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     2281 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     2341 caatcatgat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     2401 tatttagaaa aataaacaaa taggtcatga ccaaaatccc ttaacgtgag ttttcgttcc
     2461 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     2521 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     2581 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     2641 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     2701 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     2761 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     2821 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     2881 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     2941 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     3001 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     3061 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     3121 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     3181 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     3241 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
     3301 atctgtgcgg tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc
     3361 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga
     3421 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     3481 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     3541 aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg
     3601 tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt
     3661 ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg
     3721 taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg
     3781 atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg
     3841 gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt
     3901 aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac
     3961 ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag
     4021 accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc
     4081 tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc
     4141 ctcaacgaca ggagcacgat catgctagtc atgccccgcg cccaccggaa ggagctgact
     4201 gggttgaagg ctctcaaggg catcggtcga gatcccggtg cctaatgagt gagctaactt
     4261 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     4321 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt
     4381 ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct ggccctgaga
     4441 gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt
     4501 ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat
     4561 gtccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg
     4621 atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg
     4681 ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt
     4741 gcgagtgaga tatttatgcc aaccagccag acgcagacgc gccgagacag aacttaatgg
     4801 gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg
     4861 cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag
     4921 aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag
     4981 cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt
     5041 acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc
     5101 ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt
     5161 ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat
     5221 gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct
     5281 ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc
     5341 gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca
     5401 tgccataccg cgaaaggttt tgcgccattc gatggtgtcc gggatctcga cgctctccct
     5461 tatgcgactc ctgcattagg aagcagccca gtagtaggtt gaggccgttg agcaccgccg
     5521 ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag tcccccggcc acggggcctg
     5581 ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc
     5641 catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg
     5701 ccacgatgcg tccggcgtag aggatcgaga tcgatctcga tcccgcgaaa tttttccgag
     5761 gtccttgttg cgaagattga tgacaatgtg agtgcttccc ttgaaaccct gaaactgatc
     5821 cccataataa gcgaagttag cgagatgaat gcgaaaaaaa cgcggagaaa ttcatgggca
     5881 gcagccatca ccatcatcac cacagccagg atcca
//
