LOCUS       sequence-205132-        5736 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase Reporter for Formaldehyde Sensing.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5736)
  AUTHORS   Woolston BM, Roth T, Kohale I, Liu D, Stephanopoulos G
  TITLE     Development of a Formaldehyde Biosensor with Application to 
            Synthetic Methylotrophy.
  JOURNAL   Biotechnol Bioeng. 2017 Sep 16. doi: 10.1002/bit.26455.
  PUBMED    28921510
REFERENCE   2  (bases 1 to 5736)
  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: "Biotechnol 
            Bioeng. 2017 Sep 16. doi: 10.1002/bit.26455."
COMMENT     Sequence Label: pTR47m4
FEATURES             Location/Qualifiers
     source          1..5736
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             590..1669
                     /codon_start=1
                     /gene="luxA"
                     /product="LuxA luciferase subunit"
                     /label=LuxA
                     /note="from the Photorhabdus luminescens bioluminescence 
                     operon"
                     /translation="MKFGNFLLTYQPPQFSQTEVMKRLVKLGRISEECGFDTVWLLEHH
                     FTEFGLLGNPYVAAAYLLGATKKLNVGTAAIVLPTAHPVRQLEDVNLLDQMSKGRFRFG
                     ICRGLYNKDFRVFGTDMNNSRALAECWYGLIKNGMTEGYMEADNEHIKFHKVKVNPAAY
                     SRGGAPVYVVAESASTTEWAAQFGLPMILSWIINTNEKKAQLELYNEVAQEYGHDIHNI
                     DHCLSYITSVDHDSIKAKEICRKFLGHWYDSYVNATTIFDDSDQTRGYDFNKGQWRDFV
                     LKGHKDTNRRIDYSYEINPVGTPQECIDIIQKDIDATGISNICCGFEANGTVDEIIASM
                     KLFQSDVMPFLKEKQRSLLY"
     CDS             1733..2713
                     /codon_start=1
                     /gene="luxB"
                     /product="LuxB luciferase subunit"
                     /label=LuxB
                     /note="from the Photorhabdus luminescens bioluminescence 
                     operon"
                     /translation="KFGLFFLNFINSTTVQEQSIVRMQEITEYVDKLNFEQILVYENHF
                     SDNGVVGAPLTVSGFLLGLTEKIKIGSLNHIITTHHPVRIAEEACLLDQLSEGRFILGF
                     SDCEKKDEMHFFNRPVEYQQQLFEECYEIINDALTTGYCNPDNDFYSFPKISVNPHAYT
                     PGGPRKYVTATSHHIVEWAAKKGIPLIFKWDDSNDVRYEYAERYKAVADKYDVDLSEID
                     HQLMILVNYNEDSNKAKQETRAFISDYVLEMHPNENFENKLEEIIAENAVGNYTECITA
                     AKLAIEKCGAKSVLLSFEPMNDLMSQKNVINIVDDNIKKYHTEYT"
     promoter        3158..3212
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    3159..3177
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3189..3207
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    3213..3231
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             complement(join(3357..4148,4149..4217))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNKAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3980..3999
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4218..4322)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4390..4408
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(4429..5379)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      5427..5649
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 cttaagtagc accctcgcaa gctcgggcaa atcgctgaat attccttttg tctccgacca
       61 tcaggcacct gagtcgctgt ctttttcgtg acattcagtt cgctgcgctc acggctctgg
      121 cagtgaatgg gggtaaatgg cactacaggc gccttttatg gattcatgca aggaaactac
      181 ccataataca agaaaagccc gtcacgggct tctcagggcg ttttatggcg ggtctgctat
      241 gtggtgctat ctgacttttt gctgttcagc agttcctgcc ctctgatttt ccagtctgac
      301 cacttcggat tatcccgtga caggtcattc agactggcta atgcacccag taaggcagcg
      361 gtatcatcaa caggcttacc cgtcttactg tcaagaggac atccggtcaa ataaaacgaa
      421 aggctcagtc gaaagactgg gcctttcgtt ttgctgagga gacttaggga ccctaaattt
      481 tattctccag tgttatatac tatagggggg tatgcattga catatagcat acactcctat
      541 agtatattgc gtgcagataa cctgcaggtg cagtaaggag gaaaaaaaaa tgaaatttgg
      601 aaactttttg cttacatacc aacctcccca attttcccaa acagaggtaa tgaaacgttt
      661 ggttaaatta ggtcgcatct ctgaggagtg tggttttgat accgtatggt tactggagca
      721 tcatttcacg gagtttggtt tgcttggtaa cccttatgtc gctgctgcat atttacttgg
      781 cgcgactaaa aaattgaatg taggaactgc cgctattgtt cttcccacag cccatccagt
      841 acgccaactt gaagatgtga atttattgga tcaaatgtca aaaggacgat ttcggtttgg
      901 tatttgccga gggctttaca acaaggactt tcgcgtattc ggcacagata tgaataacag
      961 tcgcgcctta gcggaatgct ggtacgggct gataaagaat ggcatgacag agggatatat
     1021 ggaagctgat aatgaacata tcaagttcca taaggtaaaa gtaaaccccg cggcgtatag
     1081 cagaggtggc gcaccggttt atgtggtggc tgaatcagct tcgacgactg agtgggctgc
     1141 tcaatttggc ctaccgatga tattaagttg gattataaat actaacgaaa agaaagcaca
     1201 acttgagctt tataatgaag tggctcaaga atatgggcac gatattcata atatcgacca
     1261 ttgcttatca tatataacat ctgtagatca tgactcaatt aaagcgaaag agatttgccg
     1321 gaaatttctg gggcattggt atgattctta tgtgaatgct acgactattt ttgatgattc
     1381 agaccaaaca agaggttatg atttcaataa agggcagtgg cgtgactttg tattaaaagg
     1441 acataaagat actaatcgcc gtattgatta cagttacgaa atcaatcccg tgggaacgcc
     1501 gcaggaatgt attgacataa ttcaaaaaga cattgatgct acaggaatat caaatatttg
     1561 ttgtggattt gaagctaatg gaacagtaga cgaaattatt gcttccatga agctcttcca
     1621 gtctgatgtc atgccatttc ttaaagaaaa acaacgttcg ctattatatt atggcggtgg
     1681 cggtagcggc ggtggcggta gcggcggtgg cggtagcggc ggtggcggta gcaaatttgg
     1741 attgttcttc cttaacttca tcaattcaac aactgttcaa gaacagagta tagttcgcat
     1801 gcaggaaata acggagtatg ttgataagtt gaattttgaa cagattttag tgtatgaaaa
     1861 tcatttttca gataatggtg ttgtcggcgc tcctctgact gtttctggtt ttctgctcgg
     1921 tttaacagag aaaattaaaa ttggttcatt aaatcacatc attacaactc atcatcctgt
     1981 ccgcatagcg gaggaagctt gcttattgga tcagttaagt gaagggagat ttattttagg
     2041 gtttagtgat tgcgaaaaaa aagatgaaat gcattttttt aatcgcccgg ttgaatatca
     2101 acagcaacta tttgaagagt gttatgaaat cattaacgat gctttaacaa caggctattg
     2161 taatccagat aacgattttt atagcttccc taaaatatct gtaaatcccc atgcttatac
     2221 gccaggcgga cctcggaaat atgtaacagc aaccagtcat catattgttg agtgggcggc
     2281 caaaaaaggt attcctctca tctttaagtg ggatgattct aatgatgtta gatatgaata
     2341 tgctgaaaga tataaagccg ttgcggataa atatgacgtt gacctatcag agatagacca
     2401 tcagttaatg atattagtta actataacga agatagtaat aaagctaaac aagagacgcg
     2461 tgcatttatt agtgattatg ttcttgaaat gcaccctaat gaaaatttcg aaaataaact
     2521 tgaagaaata attgcagaaa acgctgtcgg aaattatacg gagtgtataa ctgcggctaa
     2581 gttggcaatt gaaaagtgtg gtgcgaaaag tgtattgctg tcctttgaac caatgaatga
     2641 tttgatgagc caaaaaaatg taatcaatat tgttgatgat aatattaaga agtaccacac
     2701 ggaatatacc taaacggagc caatgtacgc aaaaaacccc gcttcagcgg ggttttttcg
     2761 ccaaaaaaaa ccccgccctg tcaggggcgg ggtttttttt tagggcccta ctgactgtct
     2821 atttaagata ggctcgaacc agttcaatag tgtcgtcaac ggattggctg acttcgcggc
     2881 tgtagcagtc atttcggtca aacgtttccc ggatatggct ttcaagcact tctgccatca
     2941 gcccattagc cgcgccccga acggcagcga tctgttggag tatggcacgg cattcggcat
     3001 caccctccag cgaccgttcc agagcatcaa tctgccctcg aatacgacga actcgagtaa
     3061 ggaccttttt cttctcttcc ggagtactgg gcattttttt ttcctcctta taagggggta
     3121 gggggtctag atttttgtcg aactattcat ttcacttttc tctatcactg atagggagtg
     3181 gtaaaataac tctatcaatg atagagtgtc aatctctatc actgataggg atcatctggc
     3241 cattcgatgg acttaattaa cggcactgcg aaaaaacccc gccgaagcgg ggttttttgc
     3301 gaaatataat gaccctcttg ataacccaag agggcatttt ttaatgccca tggcgtttac
     3361 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     3421 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     3481 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     3541 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     3601 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     3661 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc tttattcagc
     3721 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     3781 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     3841 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     3901 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     3961 tgcccagcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     4021 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     4081 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     4141 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     4201 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     4261 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     4321 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     4381 acctataaaa ataggcgtat cacgaggccc ttaggccaaa taggccgttc agatccttcc
     4441 gtatttagcc agtatgttct ctagtgtggt tcgttgtttt tgcgtgagcc atgagaacga
     4501 accattgaga tcatgcttac tttgcatgtc actcaaaaat tttgcctcaa aactggtgag
     4561 ctgaattttt gcagttaaag catcgtgtag tgtttttctt agtccgttac gtaggtagga
     4621 atctgatgta atggttgttg gtattttgtc accattcatt tttatctggt tgttctcaag
     4681 ttcagttacg agatccattt gtctatctag ttcaacttgg aaaatcaacg tatcagtcgg
     4741 gcggcctcgc ttatcaacca ccaatttcat attgctgtaa gtgtttaaat ctttacttat
     4801 tggtttcaaa acccattggt taagcctttt aaactcatgg tagttatttt caagcattaa
     4861 catgaactta aattcatcaa ggctaatctc tatatttgcc ttgtgagttt tcttttgtgt
     4921 tagttctttt aataaccact cataaatcct catagagtat ttgttttcaa aagacttaac
     4981 atgttccaga ttatatttta tgaatttttt taactggaaa agataaggca atatctcttc
     5041 actaaaaact aattctaatt tttcgcttga gaacttggca tagtttgtcc actggaaaat
     5101 ctcaaagcct ttaaccaaag gattcctgat ttccacagtt ctcgtcatca gctctctggt
     5161 tgctttagct aatacaccat aagcattttc cctactgatg ttcatcatct gagcgtattg
     5221 gttataagtg aacgataccg tccgttcttt ccttgtaggg ttttcaatcg tggggttgag
     5281 tagtgccaca cagcataaaa ttagcttggt ttcatgctcc gttaagtcat agcgactaat
     5341 cgctagttca tttgctttga aaacaactaa ttcagacata catctcaatt ggtctaggta
     5401 attttaatca ctataccaat tgagatgggc tagtcaatga taattactag tccttttcct
     5461 ttgagttgtg ggtatctgta aattctgcta gacctttgct ggaaaacttg taaattctgc
     5521 tagaccctct gtaaattccg ctagaccttt gtgtgttttt tttgtttata ttcaagtggt
     5581 tataatttat agaataaaga aagaataaaa aaagataaaa agaatagatc ccagccctgt
     5641 gtataactca ctactttagt cagttccgca gtattacaaa aggatgtcgc aaacgctgtt
     5701 tgctcctcta caaaacagac cttaaaaccc taaagg
//
