LOCUS       sequence-173060-        4557 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mWasabi reporter gene driven by TlpA promoter with scrambled Pribnow
            box.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4557)
  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 4557)
  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: pTlpAsp-Wasabi
FEATURES             Location/Qualifiers
     source          1..4557
                     /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             777..821
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(859..877)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      873..920
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      957..1412
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1044..1063)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1254..1275
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(1527..2318,2319..2387))
                     /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     2150..2169
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2388..2480)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2561..3149
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3050..3069
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3303..3320
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3335..3474
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3400..3419)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3560..3582
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3576..3767)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     RBS             4487..4509
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4528..4545
                     /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 tgagctcgag tctggtaaag
      781 aaaccgctgc tgcgaaattt gaacgccagc acatggactc gtctactagc gcagcttaat
      841 taacctaggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac
      901 gggtcttgag gggttttttg ctgaaaggag gaactatatc cggattggcg aatgggacgc
      961 gccctgtagc tgcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     1021 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     1081 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     1141 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     1201 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     1261 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     1321 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     1381 gaattttaac aaaatattaa cgtttacaat ttctggcggc acgatggcat gagattatca
     1441 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     1501 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     1561 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     1621 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     1681 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     1741 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     1801 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     1861 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     1921 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     1981 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     2041 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     2101 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     2161 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     2221 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     2281 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     2341 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     2401 caatcatgat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     2461 tatttagaaa aataaacaaa taggtcatga ccaaaatccc ttaacgtgag ttttcgttcc
     2521 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     2581 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     2641 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     2701 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     2761 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     2821 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     2881 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     2941 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     3001 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     3061 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     3121 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     3181 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     3241 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     3301 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
     3361 atctgtgcgg tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc
     3421 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga
     3481 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     3541 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     3601 aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg
     3661 tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt
     3721 ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg
     3781 taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg
     3841 atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg
     3901 gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt
     3961 aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac
     4021 ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag
     4081 accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc
     4141 tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc
     4201 ctcaacgaca ggagcacgat catgctagtc atgccccgcg cccaccggaa ggagctgact
     4261 gggttgaagg ctctcaaggg catcggtcga gatcccggtg cctaatgagt gagctaactt
     4321 acattaattg cgttgcgcgc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt
     4381 ccggcgtaga ggatcgagat cgatctcgat cccgcgaaat tttaatttgt ttgttagtta
     4441 gtttatttgt tggtttgttt gtgtatttaa atcctctaga aataattttg tttaacttta
     4501 agaaggagat ataccatggg cagcagccat caccatcatc accacagcca ggatcca
//
