LOCUS       sequence-197007-        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             complement(1..951)
                     /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      999..1221
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             1898..2977
                     /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             3041..4021
                     /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        4466..4520
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    4467..4485
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4497..4515
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    4521..4539
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             complement(join(4665..5456,5457..5525))
                     /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     5288..5307
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5526..5630)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5698..5716
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcagatcctt ccgtatttag ccagtatgtt ctctagtgtg gttcgttgtt tttgcgtgag
       61 ccatgagaac gaaccattga gatcatgctt actttgcatg tcactcaaaa attttgcctc
      121 aaaactggtg agctgaattt ttgcagttaa agcatcgtgt agtgtttttc ttagtccgtt
      181 acgtaggtag gaatctgatg taatggttgt tggtattttg tcaccattca tttttatctg
      241 gttgttctca agttcggtta cgagatccat ttgtctatct agttcaactt ggaaaatcaa
      301 cgtatcagtc gggcggcctc gcttatcaac caccaatttc atattgctgt aagtgtttaa
      361 atctttactt attggtttca aaacccattg gttaagcctt ttaaactcat ggtagttatt
      421 ttcaagcatt aacatgaact taaattcatc aaggctaatc tctatatttg ccttgtgagt
      481 tttcttttgt gttagttctt ttaataacca ctcataaatc ctcatagagt atttgttttc
      541 aaaagactta acatgttcca gattatattt tatgaatttt tttaactgga aaagataagg
      601 caatatctct tcactaaaaa ctaattctaa tttttcgctt gagaacttgg catagtttgt
      661 ccactggaaa atctcaaagc ctttaaccaa aggattcctg atttccacag ttctcgtcat
      721 cagctctctg gttgctttag ctaatacacc ataagcattt tccctactga tgttcatcat
      781 ctgagcgtat tggttataag tgaacgatac cgtccgttct ttccttgtag ggttttcaat
      841 cgtggggttg agtagtgcca cacagcataa aattagcttg gtttcatgct ccgttaagtc
      901 atagcgacta atcgctagtt catttgcttt gaaaacaact aattcagaca tacatctcaa
      961 ttggtctagg tgattttaat cactatacca attgagatgg gctagtcaat gataattact
     1021 agtccttttc ctttgagttg tgggtatctg taaattctgc tagacctttg ctggaaaact
     1081 tgtaaattct gctagaccct ctgtaaattc cgctagacct ttgtgtgttt tttttgttta
     1141 tattcaagtg gttataattt atagaataaa gaaagaataa aaaaagataa aaagaataga
     1201 tcccagccct gtgtataact cactacttta gtcagttccg cagtattaca aaaggatgtc
     1261 gcaaacgctg tttgctcctc tacaaaacag accttaaaac cctaaaggct taagtagcac
     1321 cctcgcaagc tcgggcaaat cgctgaatat tccttttgtc tccgaccatc aggcacctga
     1381 gtcgctgtct ttttcgtgac attcagttcg ctgcgctcac ggctctggca gtgaatgggg
     1441 gtaaatggca ctacaggcgc cttttatgga ttcatgcaag gaaactaccc ataatacaag
     1501 aaaagcccgt cacgggcttc tcagggcgtt ttatggcggg tctgctatgt ggtgctatct
     1561 gactttttgc tgttcagcag ttcctgccct ctgattttcc agtctgacca cttcggatta
     1621 tcccgtgaca ggtcattcag actggctaat gcacccagta aggcagcggt atcatcaaca
     1681 ggcttacccg tcttactgtc aagaggacat ccggtcaaat aaaacgaaag gctcagtcga
     1741 aagactgggc ctttcgtttt gctgaggaga cttagggacc ctaaatttta ttctccagtg
     1801 ttatatacta taggggggta tgcattgaca tatagcatac actcctatag tatattgcgt
     1861 gcagataacc tgcaggtgca gtaaggagga aaaaaaaatg aaatttggaa actttttgct
     1921 tacataccaa cctccccaat tttcccaaac agaggtaatg aaacgtttgg ttaaattagg
     1981 tcgcatctct gaggagtgtg gttttgatac cgtatggtta ctggagcatc atttcacgga
     2041 gtttggtttg cttggtaacc cttatgtcgc tgctgcatat ttacttggcg cgactaaaaa
     2101 attgaatgta ggaactgccg ctattgttct tcccacagcc catccagtac gccaacttga
     2161 agatgtgaat ttattggatc aaatgtcaaa aggacgattt cggtttggta tttgccgagg
     2221 gctttacaac aaggactttc gcgtattcgg cacagatatg aataacagtc gcgccttagc
     2281 ggaatgctgg tacgggctga taaagaatgg catgacagag ggatatatgg aagctgataa
     2341 tgaacatatc aagttccata aggtaaaagt aaaccccgcg gcgtatagca gaggtggcgc
     2401 accggtttat gtggtggctg aatcagcttc gacgactgag tgggctgctc aatttggcct
     2461 accgatgata ttaagttgga ttataaatac taacgaaaag aaagcacaac ttgagcttta
     2521 taatgaagtg gctcaagaat atgggcacga tattcataat atcgaccatt gcttatcata
     2581 tataacatct gtagatcatg actcaattaa agcgaaagag atttgccgga aatttctggg
     2641 gcattggtat gattcttatg tgaatgctac gactattttt gatgattcag accaaacaag
     2701 aggttatgat ttcaataaag ggcagtggcg tgactttgta ttaaaaggac ataaagatac
     2761 taatcgccgt attgattaca gttacgaaat caatcccgtg ggaacgccgc aggaatgtat
     2821 tgacataatt caaaaagaca ttgatgctac aggaatatca aatatttgtt gtggatttga
     2881 agctaatgga acagtagacg aaattattgc ttccatgaag ctcttccagt ctgatgtcat
     2941 gccatttctt aaagaaaaac aacgttcgct attatattat ggcggtggcg gtagcggcgg
     3001 tggcggtagc ggcggtggcg gtagcggcgg tggcggtagc aaatttggat tgttcttcct
     3061 taacttcatc aattcaacaa ctgttcaaga acagagtata gttcgcatgc aggaaataac
     3121 ggagtatgtt gataagttga attttgaaca gattttagtg tatgaaaatc atttttcaga
     3181 taatggtgtt gtcggcgctc ctctgactgt ttctggtttt ctgctcggtt taacagagaa
     3241 aattaaaatt ggttcattaa atcacatcat tacaactcat catcctgtcc gcatagcgga
     3301 ggaagcttgc ttattggatc agttaagtga agggagattt attttagggt ttagtgattg
     3361 cgaaaaaaaa gatgaaatgc atttttttaa tcgcccggtt gaatatcaac agcaactatt
     3421 tgaagagtgt tatgaaatca ttaacgatgc tttaacaaca ggctattgta atccagataa
     3481 cgatttttat agcttcccta aaatatctgt aaatccccat gcttatacgc caggcggacc
     3541 tcggaaatat gtaacagcaa ccagtcatca tattgttgag tgggcggcca aaaaaggtat
     3601 tcctctcatc tttaagtggg atgattctaa tgatgttaga tatgaatatg ctgaaagata
     3661 taaagccgtt gcggataaat atgacgttga cctatcagag atagaccatc agttaatgat
     3721 attagttaac tataacgaag atagtaataa agctaaacaa gagacgcgtg catttattag
     3781 tgattatgtt cttgaaatgc accctaatga aaatttcgaa aataaacttg aagaaataat
     3841 tgcagaaaac gctgtcggaa attatacgga gtgtataact gcggctaagt tggcaattga
     3901 aaagtgtggt gcgaaaagtg tattgctgtc ctttgaacca atgaatgatt tgatgagcca
     3961 aaaaaatgta atcaatattg ttgatgataa tattaagaag taccacacgg aatataccta
     4021 aacggagcca atgtacgcaa aaaaccccgc ttcagcgggg ttttttcgcc aaaaaaaacc
     4081 ccgccctgtc aggggcgggg ttttttttta gggccctact gactgtctat ttaagatagg
     4141 ctcgaaccag ttcaatagtg tcgtcaacgg attggctgac ttcgcggctg tagcagtcat
     4201 ttcggtcaaa cgtttcccgg atatggcttt caagcacttc tgccatcagc ccattagccg
     4261 cgccccgaac ggcagcgatc tgttggagta tggcacggca ttcggcatca ccctccagcg
     4321 accgttccag agcatcaatc tgccctcgaa tacgacgaac tcgagtaagg acctttttct
     4381 tctcttccgg agtactgggc attttttttt cctccttata agggggtagg gggtctagat
     4441 ttttgtcgaa ctattcattt cacttttctc tatcactgat agggagtggt aaaataactc
     4501 tatcaatgat agagtgtcaa tctctatcac tgatagggat catctggcca ttcgatggac
     4561 ttaattaacg gcactgcgaa aaaaccccgc cgaagcgggg ttttttgcga aatataatga
     4621 ccctcttgat aacccaagag ggcatttttt aatgcccatg gcgtttacca atgcttaatc
     4681 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     4741 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     4801 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     4861 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     4921 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     4981 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     5041 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     5101 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     5161 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     5221 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     5281 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     5341 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     5401 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     5461 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     5521 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     5581 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     5641 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     5701 aggcgtatca cgaggccctt aggccaaata ggccgt
//
