LOCUS       sequence-225099-        5768 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding the bacterial expression of the control Snifit-MTX 
            (FRET semi-synthetic biosensor).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5768)
  AUTHORS   Xue L, Prifti E, Johnsson K
  TITLE     A General Strategy for the Semisynthesis of Ratiometric Fluorescent 
            Sensor Proteins with Increased Dynamic Range.
  JOURNAL   J Am Chem Soc. 2016 Apr 27;138(16):5258-61. doi: 
            10.1021/jacs.6b03034. Epub 2016 Apr 15.
  PUBMED    27071001
REFERENCE   2  (bases 1 to 5768)
  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: 
            "10.1021/jacs.6b03034"; journalName: "J Am Chem Soc"; date: 
            "2016-04-27- 27"; volume: "138"; issue: "16"; pages: "5258-61"
COMMENT     Sequence Label: pBAD-MTX_Snifit
FEATURES             Location/Qualifiers
     source          1..5768
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        17..301
                     /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     224..243
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             344..367
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             374..388
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     CDS             1601..2077
                     /codon_start=1
                     /gene="folA"
                     /product="E. coli dihydrofolate reductase"
                     /label=ecDHFR
                     /translation="MISLIAALAVDRVIGMENAMPWNLPADLAWFKRNTLNKPVIMGLQ
                     TWESIGRPLPGRKNIILSSQPGTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVYEQFL
                     PKAQKLYLTHIDAEVEGDTHFPDYEPDDWESVFSEFHDADAQNSHSYCFEILERR"
     CDS             2108..2137
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(2184..2201)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2354..2440
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2532..2559
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2579..2670
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2671..2739,2740..3531)
                     /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(2889..2908)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3702..4290
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4191..4210
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4444..4461
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4476..4616
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4542..4561)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4702..4724
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4880..5758)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 ctcccgccat tcagagaaga aaccaattgt ccatattgca tcagacattg ccgtcactgc
       61 gtcttttact ggctcttctc gctaaccaaa ccggtaaccc cgcttattaa aagcattctg
      121 taacaaagcg ggaccaaagc catgacaaaa acgcgtaaca aaagtgtcta taatcacggc
      181 agaaaagtcc acattgatta tttgcacggc gtcacacttt gctatgccat agcattttta
      241 tccataagat tagcggatcc tacctgacgc tttttatcgc aactctctac tgtttctcca
      301 tacccgtttt ttgggctaac aggaggaatt aaccatggca agctggagcc acccgcagtt
      361 cgaaaagggt gcagatgacg acgacaaggt accgcatatg gataaagact gcgaaatgaa
      421 acgcactacc ctggacagcc cgctgggtaa actggaactg agcggttgcg agcagggcct
      481 gcacgaaatc attttcctgg gcaaaggtac gagcgctgcg gatgcggtcg aagtaccggc
      541 tccggctgcg gttctgggtg gcccggagcc gctgatgcag gctaccgcat ggctgaacgc
      601 atattttcac cagccagagg cgattgaaga gttccctgta ccggccctgc atcacccggt
      661 atttcagcag gaatccttta cccgtcaggt actgtggaaa ctgctgaaag tagtgaaatt
      721 cggcgaggtg atctcctatt cccatctggc ggcgctggct ggcaacccag cggcaactgc
      781 ggcggtgaaa accgctctgt ctggtaatcc ggtgccaatt ctgatcccgt gccaccgtgt
      841 ggttcagggc gacctggacg taggtggcta cgaaggcggc ctggctgtga aagaatggct
      901 gctggcgcac gaaggtcacc gtctgggtaa accaggcctg ggcgaattcc ctcctcctcc
      961 gccaccgcca ccaccgccgc cgccgccacc accacctcct ccgcctcctc caccaccacc
     1021 gccaccgcct ccaccacctg gcggccgcat ggacaaggat tgtgaaatga aacgcaccac
     1081 actggacagc cctttgggga agctggagct gtctggttgt gagcagggtc tgcacaaaat
     1141 aatttttctg ggcaagggga cgtctgcagc tgatgccgtg gaggtcccag cccccgctgc
     1201 ggttctcgga ggtccggagc ccctgataca ggctacagcc tggctgaatg cctatttcca
     1261 ccagcccgag gctatcgaag agttccccgt gccggctctt caccatcccg ttttccagca
     1321 agagtcgttc accagacagg tgttatggaa gctgctgaag gttgtgaaat tcggagaagt
     1381 gatttctgag tcacacttag cagccctggt aggcaacccc gcagccacgg cagcagtgaa
     1441 tacggcactg gatggcaatc ctgtccctat cctgatcccg tgccacagag tggtccaggg
     1501 ggattcggat gtggggccgt accttggtgg actggccgtg aaggaatggc ttctggccca
     1561 tgaaggccac cggttgggga agccaggctt gggactcgag atgatcagtc tgattgcggc
     1621 gttagcggta gatcgcgtta tcggcatgga aaacgccatg ccgtggaacc tgcctgccga
     1681 tctcgcctgg tttaaacgca acaccttaaa taaacccgtg attatgggcc tccaaacctg
     1741 ggaatcaatc ggtcgtccgt tgccaggacg caaaaatatt atcctcagca gtcaaccggg
     1801 tacggacgat cgcgtaacgt gggtgaagtc ggtggatgaa gccatcgcgg cgtgtggtga
     1861 cgtaccagaa atcatggtga ttggcggcgg tcgcgtttat gaacagttct tgccaaaagc
     1921 gcaaaaactg tatctgacgc atatcgacgc agaagtggaa ggcgacaccc atttcccgga
     1981 ttacgagccg gatgactggg aatcggtatt cagcgaattc cacgatgctg atgcgcagaa
     2041 ctctcacagc tattgctttg agattctgga gcggcgggga gctccgggct tctcctcaat
     2101 ttccgctcat caccaccatc atcaccatca ccaccactaa ctcgagagct tggctgtttt
     2161 ggcggatgag agaagatttt cagcctgata cagattaaat cagaacgcag aagcggtctg
     2221 ataaaacaga atttgcctgg cggcagtagc gcggtggtcc cacctgaccc catgccgaac
     2281 tcagaagtga aacgccgtag cgccgatggt agtgtggggt ctccccatgc gagagtaggg
     2341 aactgccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat
     2401 ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgggag cggatttgaa
     2461 cgttgcgaag caacggcccg gagggtggcg ggcaggacgc ccgccataaa ctgccaggca
     2521 tcaaattaag cagaaggcca tcctgacgga tggccttttt gcgtttctac aaactctttt
     2581 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     2641 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     2701 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     2761 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     2821 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     2881 aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt
     2941 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     3001 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     3061 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     3121 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     3181 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     3241 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     3301 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     3361 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     3421 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     3481 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     3541 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     3601 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     3661 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     3721 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     3781 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     3841 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     3901 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     3961 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4021 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4081 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4141 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     4201 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     4261 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     4321 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     4381 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     4441 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
     4501 catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc
     4561 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc
     4621 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     4681 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     4741 cgaaacgcgc gaggcagcag atcaattcgc gcgcgaaggc gaagcggcat gcataatgtg
     4801 cctgtcaaat ggacgaagca gggattctgc aaaccctatg ctactccgtc aagccgtcaa
     4861 ttgtctgatt cgttaccaat tatgacaact tgacggctac atcattcact ttttcttcac
     4921 aaccggcacg gaactcgctc gggctggccc cggtgcattt tttaaatacc cgcgagaaat
     4981 agagttgatc gtcaaaacca acattgcgac cgacggtggc gataggcatc cgggtggtgc
     5041 tcaaaagcag cttcgcctgg ctgatacgtt ggtcctcgcg ccagcttaag acgctaatcc
     5101 ctaactgctg gcggaaaaga tgtgacagac gcgacggcga caagcaaaca tgctgtgcga
     5161 cgctggcgat atcaaaattg ctgtctgcca ggtgatcgct gatgtactga caagcctcgc
     5221 gtacccgatt atccatcggt ggatggagcg actcgttaat cgcttccatg cgccgcagta
     5281 acaattgctc aagcagattt atcgccagca gctccgaata gcgcccttcc ccttgcccgg
     5341 cgttaatgat ttgcccaaac aggtcgctga aatgcggctg gtgcgcttca tccgggcgaa
     5401 agaaccccgt attggcaaat attgacggcc agttaagcca ttcatgccag taggcgcgcg
     5461 gacgaaagta aacccactgg tgataccatt cgcgagcctc cggatgacga ccgtagtgat
     5521 gaatctctcc tggcgggaac agcaaaatat cacccggtcg gcaaacaaat tctcgtccct
     5581 gatttttcac caccccctga ccgcgaatgg tgagattgag aatataacct ttcattccca
     5641 gcggtcggtc gataaaaaaa tcgagataac cgttggcctc aatcggcgtt aaacccgcca
     5701 ccagatgggc attaaacgag tatcccggca gcaggggatc attttgcgct tcagccatac
     5761 ttttcata
//
