LOCUS       sequence-173077-        7171 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Nonfluorescent variant of mWasabi reporter gene gated by wild type 
            TlpA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7171)
  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 7171)
  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: pTlpA-Wasabi-NF
FEATURES             Location/Qualifiers
     source          1..7171
                     /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
                     TNTINLEVKEGAPLPFSYDILTTAFTYANRAFTKYPDDIPNYFKQSFPEGYSWERTMTF
                     EDKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKETTGWDASTERMYVRDGVLK
                     GDVKMKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYEIA
                     VARNSTDGMDELYKG"
     promoter        768..844
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2000..2044
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2082..2100)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2096..2143
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2180..2635
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2267..2286)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2477..2498
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(2750..3541,3542..3610))
                     /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     3373..3392
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3611..3703)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3784..4372
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4273..4292
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4526..4543
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4558..4697
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4623..4642)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4783..4805
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4799..4990)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5562..6644)
                     /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     6606..6625
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6645..6722)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6928..6947
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     RBS             7101..7123
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             7142..7159
                     /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 acctacgcca 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 ctgaaagctt
      721 gcggccgcat aatgcttaag tcgaacagaa agtaatcgta tgagctcgac accatcgaat
      781 ggcgcaaacc tttcgcggta tggcatgata gcgcccggaa gagagtcaat tcagggtggt
      841 gaatgtataa gaaggagata tacatatgcg tccggcgaca tacgaaccag aacagattat
      901 tgaagcaggg ctggccctgc aggctgaagg acggaatatc accgggttcg cactacgtaa
      961 ccaggtgggt ggcggcaatc cgacacgtct ccgccagata tgggacgaat accaggcttc
     1021 acagagcacg gtcgtcactg aacccgttgc cgagctgcca gtggaagtgg ctgaagaagt
     1081 gaaggccgtc tccgccgcgc tgtccgaacg catcacccag ctggcgacag aactgaatga
     1141 caaggcggtc cgggctgcag aacgccgggt tgcggaagtc acgcgtgctg ccggtgaaca
     1201 gaccgcacag gcagagcggg agctggccga cgccgcgcag acagtcgacg acctggaaga
     1261 aaaactggat gaactgcagg acagatatga cagtttgacg ctggcgctgg agtcagaacg
     1321 ttcactgcgt cagcagcatg atgtggagat ggcccagctg aaagagcgtc ttgcggccgc
     1381 tgaagagaat acccgtcagc gagaggaacg gtatcaggag cagaagacag tgctgcagga
     1441 tgcgcttaat gcggagcagg cacagcacaa aaacacgcgg gaagacctgc agaaacgact
     1501 ggagcaaatt tctgccgaag ctaatgcgcg tacagaagaa ctgaagtctg aacgcgataa
     1561 agtcaatact ctccttaccc gccttgaatc gcaggaaaat gcgctggcct cagaacgtca
     1621 gcagcatctg gccacccgcg aaacgctgca gcaacgcctc gagcaggcca tcgctgacac
     1681 gcaggcgcgc gccggtgaga ttgcacttga acgtgacaga gtcagcagcc tcaccgcaag
     1741 gctggaatcg caggaaaagg cctcctcgga gcaactggtg cgtatgggca gtgaaatagc
     1801 cagtctgaca gagcgttgca cacagctgga aaaccagcgt gatgatgccc gtctggagac
     1861 gatgggggag aaagaaacgg tcgcggcact gcgtggtgag gctgaagccc tgaagcgtca
     1921 gaaccagtca ctgatggcgg cgctttcagg caataaacag accggtggcc agaatgcgtg
     1981 aggtaccctc gagtctggta aagaaaccgc tgctgcgaaa tttgaacgcc agcacatgga
     2041 ctcgtctact agcgcagctt aattaaccta ggctgctgcc accgctgagc aataactagc
     2101 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat
     2161 atccggattg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     2221 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     2281 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     2341 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     2401 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     2461 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     2521 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     2581 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aatttctggc
     2641 ggcacgatgg catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     2701 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     2761 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     2821 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     2881 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     2941 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     3001 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     3061 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     3121 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     3181 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     3241 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     3301 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     3361 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     3421 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     3481 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     3541 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     3601 gaatactcat actcttcctt tttcaatcat gattgaagca tttatcaggg ttattgtctc
     3661 atgagcggat acatatttga atgtatttag aaaaataaac aaataggtca tgaccaaaat
     3721 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     3781 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     3841 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     3901 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     3961 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4021 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4081 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     4141 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     4201 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     4261 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     4321 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     4381 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     4441 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     4501 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct
     4561 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcaatg gtgcactctc
     4621 agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg
     4681 actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     4741 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     4801 agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt
     4861 ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct
     4921 ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct
     4981 gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt catgggggta atgataccga
     5041 tgaaacgaga gaggatgctc acgatacggg ttactgatga tgaacatgcc cggttactgg
     5101 aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga aaaatcactc
     5161 agggtcaatg ccagcgcttc gttaatacag atgtaggtgt tccacagggt agccagcagc
     5221 atcctgcgat gcagatccgg aacataatgg tgcagggcgc tgacttccgc gtttccagac
     5281 tttacgaaac acggaaaccg aagaccattc atgttgttgc tcaggtcgca gacgttttgc
     5341 agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc attctgctaa ccagtaaggc
     5401 aaccccgcca gcctagccgg gtcctcaacg acaggagcac gatcatgcta gtcatgcccc
     5461 gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg
     5521 gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt
     5581 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     5641 tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg
     5701 cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc
     5761 aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta
     5821 tcgtcgtatc ccactaccga gatgtccgca ccaacgcgca gcccggactc ggtaatggcg
     5881 cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc
     5941 tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt
     6001 tccgctatcg gctgaatttg attgcgagtg agatatttat gccaaccagc cagacgcaga
     6061 cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg
     6121 accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg
     6181 ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca
     6241 gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg
     6301 agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc
     6361 accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc
     6421 gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt
     6481 tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc
     6541 cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag
     6601 acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat
     6661 tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg
     6721 tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc ccagtagtag
     6781 gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa
     6841 cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc
     6901 gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc
     6961 acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatcg agatcgatct
     7021 cgatcccgcg aaattttaat ttgtttgtta gttagtttat ttgttggttt gtttgtgtta
     7081 taatatcctc tagaaataat tttgtttaac tttaagaagg agatatacca tgggcagcag
     7141 ccatcaccat catcaccaca gccaggatcc a
//
