LOCUS       sequence-222071-        5265 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of TrxRFP1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5265)
  AUTHORS   Fan Y, Makar M, Wang MX, Ai HW
  TITLE     Monitoring thioredoxin redox with a genetically encoded red 
            fluorescent biosensor.
  JOURNAL   Nat Chem Biol. 2017 Jun 26. doi: 10.1038/nchembio.2417.
  PUBMED    28671680
REFERENCE   2  (bases 1 to 5265)
  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: "Nat Chem 
            Biol. 2017 Jun 26. doi: 10.1038/nchembio.2417."
COMMENT     Sequence Label: pBAD-TrxRFP1
FEATURES             Location/Qualifiers
     source          1..5265
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..285
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             331..348
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             352..384
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(388..396,397..411)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     1560..1579
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1666..1683)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1836..1922
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2014..2041
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2060..2151
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2152..2220,2221..3012)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2370..2389)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3183..3771
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3672..3691
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3925..3942
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3957..4097
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4023..4042)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4183..4205
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4361..5239)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg ttttttgggc
      301 taacaggagg aattaaccat ggggggttct catcatcatc atcatcatgg tatggctagc
      361 atgactggtg gacagcaaat gggtcgggat ctgtacgacg atgacgataa ggatccgagc
      421 tcgagcatgg ttaaacagat cgaatctaaa accgcgttcc aggaagcgct ggacgcggcg
      481 ggtgacaaac tggttgttgt tgacttctct gcgacctggt gcggtccgtg caaaatgatc
      541 aaaccgttct tccactctct gtctgaaaaa tactctaacg ttatcttcct ggaagttgac
      601 gttgacgact cccaggacgt tgcgtctgaa tccgaagtta aatccatgcc gaccttccag
      661 ttcttcaaaa aaggtcagaa agttggtgaa ttctctggtg cgaacaaaga aaaactggaa
      721 gcgaccatca acgaactggt tggtggcagt ggtggcggcg gtagtggcgg tggagggagc
      781 ggaggggggg gaagcggggg aggtggaagt ggcgggggag gtagtggggg cgagcttagt
      841 tgcgctataa gtcggctgag ctcacccgtg gtttccgagc ggatgtaccc cgaggacggc
      901 gccctgaaga gcgagatcaa gaaggggctg aggctgaagg acggcggcca ctacgccgcc
      961 gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacatcgtc
     1021 gacatcaagt tggacatcgt gtcccacaac gaggactaca ccatcgtgga acagtacgaa
     1081 cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaaggg aggtacaggc
     1141 gggagtctgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc
     1201 ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
     1261 gagggccgcc cctacgaggc ctttcagacc gctaagctga aggtgaccaa gggtggcccc
     1321 ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacatt
     1381 aagcacccag ccgacatccc cgactacttc aagctgtcct tccccgaggg cttcaggtgg
     1441 gagcgcgtga tgaacttcga ggacggcggc attattcacg ttaaccagga ctcctccctg
     1501 caggacggcg tattcatcta caaggtgaag ctgcgcggca ccaacttccc ccccgacggc
     1561 cccgtaatgc agaagaagac catgggctgg gaggctacgc gtgaccaaag gtgctccagt
     1621 gtggcttaag cttggctgtt ttggcggatg agagaagatt ttcagcctga tacagattaa
     1681 atcagaacgc agaagcggtc tgataaaaca gaatttgcct ggcggcagta gcgcggtggt
     1741 cccacctgac cccatgccga actcagaagt gaaacgccgt agcgccgatg gtagtgtggg
     1801 gtctccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga
     1861 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa
     1921 atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac
     1981 gcccgccata aactgccagg catcaaatta agcagaaggc catcctgacg gatggccttt
     2041 ttgcgtttct acaaactctt ttgtttattt ttctaaatac attcaaatat gtatccgctc
     2101 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     2161 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     2221 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     2281 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     2341 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac
     2401 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     2461 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     2521 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     2581 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     2641 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     2701 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     2761 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     2821 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     2881 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     2941 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     3001 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     3061 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     3121 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     3181 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     3241 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     3301 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     3361 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     3421 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     3481 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     3541 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     3601 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     3661 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     3721 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     3781 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     3841 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     3901 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg
     3961 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc
     4021 agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg
     4081 actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     4141 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     4201 agaggttttc accgtcatca ccgaaacgcg cgaggcagca gatcaattcg cgcgcgaagg
     4261 cgaagcggca tgcataatgt gcctgtcaaa tggacgaagc agggattctg caaaccctat
     4321 gctactccgt caagccgtca attgtctgat tcgttaccaa ttatgacaac ttgacggcta
     4381 catcattcac tttttcttca caaccggcac ggaactcgct cgggctggcc ccggtgcatt
     4441 ttttaaatac ccgcgagaaa tagagttgat cgtcaaaacc aacattgcga ccgacggtgg
     4501 cgataggcat ccgggtggtg ctcaaaagca gcttcgcctg gctgatacgt tggtcctcgc
     4561 gccagcttaa gacgctaatc cctaactgct ggcggaaaag atgtgacaga cgcgacggcg
     4621 acaagcaaac atgctgtgcg acgctggcga tatcaaaatt gctgtctgcc aggtgatcgc
     4681 tgatgtactg acaagcctcg cgtacccgat tatccatcgg tggatggagc gactcgttaa
     4741 tcgcttccat gcgccgcagt aacaattgct caagcagatt tatcgccagc agctccgaat
     4801 agcgcccttc cccttgcccg gcgttaatga tttgcccaaa caggtcgctg aaatgcggct
     4861 ggtgcgcttc atccgggcga aagaaccccg tattggcaaa tattgacggc cagttaagcc
     4921 attcatgcca gtaggcgcgc ggacgaaagt aaacccactg gtgataccat tcgcgagcct
     4981 ccggatgacg accgtagtga tgaatctctc ctggcgggaa cagcaaaata tcacccggtc
     5041 ggcaaacaaa ttctcgtccc tgatttttca ccaccccctg accgcgaatg gtgagattga
     5101 gaatataacc tttcattccc agcggtcggt cgataaaaaa atcgagataa ccgttggcct
     5161 caatcggcgt taaacccgcc accagatggg cattaaacga gtatcccggc agcaggggat
     5221 cattttgcgc ttcagccata cttttcatac tcccgccatt cagag
//
