LOCUS       sequence-323558-        7172 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mWasabi reporter gene gated by TlpA39.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7172)
  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 7172)
  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: pTlpA39-Wasabi
FEATURES             Location/Qualifiers
     source          1..7172
                     /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"
     promoter        768..845
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2001..2045
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2083..2101)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2097..2144
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2181..2636
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2268..2287)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2478..2499
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(2751..3542,3543..3611))
                     /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     3374..3393
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3612..3704)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3785..4373
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4274..4293
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4527..4544
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4559..4698
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4624..4643)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4784..4806
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4800..4991)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5563..6645)
                     /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     6607..6626
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6646..6723)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6929..6948
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     RBS             7102..7124
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             7143..7160
                     /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 ctgaaagctt
      721 gcggccgcat aatgcttaag tcgaacagaa agtaatcgta tgagctcgac accatcgaat
      781 ggcgcaaaac ctttcgcggt atggcatgat agcgcccgga agagagtcaa ttcagggtgg
      841 tgaatgtata agaaggagat atacatatgc gtccggcgac atacgaacca gaacagatta
      901 ttgaagcagg gctggccctg caggctgaag gacggaatat caccgggttc gcactacgta
      961 accaggtggg tggcggcaat ccgacacgtc tccgccagat atgggacgaa taccaggctt
     1021 cacagagcac ggtcgtcact gaacccgttg ccgagctgcc agtggaagtg gctgaagaag
     1081 tgaaggccgt ctccgccgcg ctgtccgaac gcatcaccca gctggcgaca gaactgaatg
     1141 acaaggcggt ccgggctgca gaacgccggg ttgcggaagt cacgcgtgct gccggtgaac
     1201 agaccgcaca ggcagagcgg gagctggccg acgccgcgca gacagtcgac gacctggaag
     1261 aaaaactggt tgaactgcag gacagatatg acagtttgac gctggcgctg gagtcagaac
     1321 gttcactgcg tcagcagcat gatgtggaga tggcccagct gaaagagcgt cttgcggccg
     1381 ctgaagagaa tacccgtcag cgagaggaac ggtatcagga gcagaagaca gtgctgcagg
     1441 atgcgcttaa tgcggagcag gcacagcaca aaaacacgcg ggaagacctg cagaaacgac
     1501 tggagcaaat ttctgtcgaa gctaatgcgc gtacagaaga actgaagtct gaacgcgata
     1561 aagtcaatac tttccttacc cgccttgaat cgcaggaaaa tgcgctggcc tcagaacgtc
     1621 agcagcatct ggccacccgc gaaacgctgc agcaacgcct cgagcaggcc atcgctgaca
     1681 cgcaggcgcg cgccggtgag attgcacttg aacgtgacag agtcagcagc ctcaccgcaa
     1741 ggctggaatc gcaggaaaag gcctcctcgg agcaactggt gcgtatgggc agtgaaatag
     1801 ccagtctgac agagcgttgc acacagctgg aaaaccagcg tgatgatgcc cgtctggaga
     1861 cgatggggga gaaagaaacg gtcgcggcac tgcgtggtga ggctgaagcc ctgaagcgtc
     1921 agaaccagtc actgatggcg gcgctttcag gcaataaaca gaccggtggc cagaatgcgt
     1981 gaggtaccct cgagtctggt aaagaaaccg ctgctgcgaa atttgaacgc cagcacatgg
     2041 actcgtctac tagcgcagct taattaacct aggctgctgc caccgctgag caataactag
     2101 cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta
     2161 tatccggatt ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2221 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     2281 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     2341 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     2401 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     2461 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     2521 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     2581 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caatttctgg
     2641 cggcacgatg gcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     2701 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     2761 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     2821 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     2881 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     2941 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     3001 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     3061 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     3121 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     3181 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     3241 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     3301 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     3361 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     3421 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     3481 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     3541 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     3601 tgaatactca tactcttcct ttttcaatca tgattgaagc atttatcagg gttattgtct
     3661 catgagcgga tacatatttg aatgtattta gaaaaataaa caaataggtc atgaccaaaa
     3721 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     3781 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     3841 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     3901 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     3961 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     4021 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     4081 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     4141 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     4201 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     4261 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     4321 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     4381 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     4441 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     4501 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     4561 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcaat ggtgcactct
     4621 cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt
     4681 gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     4741 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     4801 cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg
     4861 tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc
     4921 tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc
     4981 tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg
     5041 atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg
     5101 gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact
     5161 cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag
     5221 catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga
     5281 ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg
     5341 cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg
     5401 caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgct agtcatgccc
     5461 cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg tcgagatccc
     5521 ggtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag
     5581 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     5641 ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt
     5701 gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag
     5761 caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt
     5821 atcgtcgtat cccactaccg agatgtccgc accaacgcgc agcccggact cggtaatggc
     5881 gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc
     5941 ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg
     6001 ttccgctatc ggctgaattt gattgcgagt gagatattta tgccaaccag ccagacgcag
     6061 acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc
     6121 gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat
     6181 gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac
     6241 agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc
     6301 gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac
     6361 caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg
     6421 cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag
     6481 ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc
     6541 ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga
     6601 gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa
     6661 ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc attcgatggt
     6721 gtccgggatc tcgacgctct cccttatgcg actcctgcat taggaagcag cccagtagta
     6781 ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag atggcgccca
     6841 acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc
     6901 cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg
     6961 cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggatc gagatcgatc
     7021 tcgatcccgc gaaattttaa tttgtttgtt agttagttta tttgttggtt tgtttgtgtt
     7081 ataatatcct ctagaaataa ttttgtttaa ctttaagaag gagatatacc atgggcagca
     7141 gccatcacca tcatcaccac agccaggatc ca
//
