LOCUS       sequence-197581-        4295 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses sfGFP from the OmpR regulated PompF promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4295)
  AUTHORS   Landry BP, Palanki R, Dyulgyarov N, Hartsough LA, Tabor JJ
  TITLE     Phosphatase activity tunes two-component system sensor detection 
            threshold.
  JOURNAL   Nat Commun. 2018 Apr 12;9(1):1433. doi: 10.1038/s41467-018-03929-y.
  PUBMED    29650958
REFERENCE   2  (bases 1 to 4295)
  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 
            Commun."; date: "2018-04-12"; volume: "9(1):1433. doi"; pages: " 
            10.1038/s41467-018-03929-y"
COMMENT     Sequence Label: pRS52
FEATURES             Location/Qualifiers
     source          1..4295
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1..58
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(82..99)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(361..1311)
                     /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      1359..1581
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(2073..2167)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(2197..2988,2989..3057))
                     /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     2820..2839
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3058..3162)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3227..3245
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(3249..3292)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     CDS             3441..4154
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      4175..4246
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4262..4289
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 tccggcaaaa aagggcaagg tgtcaccacc ctgccctttt tctttaaaac cgaaaagatt
       61 acttcgcgtt atgcaggctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
      121 gcgagcggta tcagctcact caaaggcggt aatctcgagg ttacattgtc gatctgttca
      181 tggtgaacag ctttaaatgc accaaaaact cgtaaaagct ctgatgtatc tatctttttt
      241 acaccgtttt catctgtgca tatggacagt tttccctttg atatctaacg gtgaacagtt
      301 gttctacttt tgtttgttag tcttgatgct tcactgatag atacaagagc cataagaacc
      361 tcagatcctt ccgtatttag ccagtatgtt ctctagtgtg gttcgttgtt tttgcgtgag
      421 ccatgagaac gaaccattga gatcatgctt actttgcatg tcactcaaaa attttgcctc
      481 aaaactggtg agctgaattt ttgcagttaa agcatcgtgt agtgtttttc ttagtccgtt
      541 acgtaggtag gaatctgatg taatggttgt tggtattttg tcaccattca tttttatctg
      601 gttgttctca agttcggtta cgagatccat ttgtctatct agttcaactt ggaaaatcaa
      661 cgtatcagtc gggcggcctc gcttatcaac caccaatttc atattgctgt aagtgtttaa
      721 atctttactt attggtttca aaacccattg gttaagcctt ttaaactcat ggtagttatt
      781 ttcaagcatt aacatgaact taaattcatc aaggctaatc tctatatttg ccttgtgagt
      841 tttcttttgt gttagttctt ttaataacca ctcataaatc ctcatagagt atttgttttc
      901 aaaagactta acatgttcca gattatattt tatgaatttt tttaactgga aaagataagg
      961 caatatctct tcactaaaaa ctaattctaa tttttcgctt gagaacttgg catagtttgt
     1021 ccactggaaa atctcaaagc ctttaaccaa aggattcctg atttccacag ttctcgtcat
     1081 cagctctctg gttgctttag ctaatacacc ataagcattt tccctactga tgttcatcat
     1141 ctgagcgtat tggttataag tgaacgatac cgtccgttct ttccttgtag ggttttcaat
     1201 cgtggggttg agtagtgcca cacagcataa aattagcttg gtttcatgct ccgttaagtc
     1261 atagcgacta atcgctagtt catttgcttt gaaaacaact aattcagaca tacatctcaa
     1321 ttggtctagg tgattttaat cactatacca attgagatgg gctagtcaat gataattaca
     1381 tgtccttttc ctttgagttg tgggtatctg taaattctgc tagacctttg ctggaaaact
     1441 tgtaaattct gctagaccct ctgtaaattc cgctagacct ttgtgtgttt tttttgttta
     1501 tattcaagtg gttataattt atagaataaa gaaagaataa aaaaagataa aaagaataga
     1561 tcccagccct gtgtataact cactacttta gtcagttccg cagtattaca aaaggatgtc
     1621 gcaaacgctg tttgctcctc tacaaaacag accttaaaac cctaaaggct taagtagcac
     1681 cctcgcaagc tcgggcaaat cgctgaatat tccttttgtc tccgaccatc aggcacctga
     1741 gtcgctgtct ttttcgtgac attcagttcg ctgcgctcac ggctctggca gtgaatgggg
     1801 gtaaatggca ctacaggcgc cttttatgga ttcatgcaag gaaactaccc ataatacaag
     1861 aaaagcccgt cacgggcttc tcagggcgtt ttatggcggg tctgctatgt ggtgctatct
     1921 gactttttgc tgttcagcag ttcctgccct ctgattttcc agtctgacca cttcggatta
     1981 tcccgtgaca ggtcattcag actggctaat gcacccagta aggcagcggt atcatcaaca
     2041 ggcttacccg tcttactgtc cctagtgctt ggattctcac caataaaaaa cgcccggcgg
     2101 caaccgagcg ttctgaacaa atccagatgg agttctgagg tcattactgg atctatcaac
     2161 aggagtccaa gcgagctcgt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     2221 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     2281 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     2341 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     2401 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     2461 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     2521 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     2581 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     2641 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     2701 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     2761 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     2821 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     2881 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     2941 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     3001 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     3061 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     3121 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc ccatggtgcc
     3181 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     3241 gaggcagaat ttcagataaa aaaaatcctt agctttcgct aaggatgatt tctggttcct
     3301 taaattttac ttttggttac atattttttc tttttgaaac caaatcttta tctttgtagc
     3361 actttcacgg tagcgaaacg ttagtttgaa tggaaagatg cctgcaggtg tcataaaaaa
     3421 aaccatgagg gtaataaata atgcgtaaag gcgaagagct gttcactggt gtcgtcccta
     3481 ttctggtgga actggatggt gatgtcaacg gtcataagtt ttccgtgcgt ggcgagggtg
     3541 aaggtgacgc aactaatggt aaactgacgc tgaagttcat ctgtactact ggtaaactgc
     3601 cggtaccttg gccgactctg gtaacgacgc tgacttatgg tgttcagtgc tttgctcgtt
     3661 atccggacca tatgaagcag catgacttct tcaagtccgc catgccggaa ggctatgtgc
     3721 aggaacgcac gatttccttt aaggatgacg gcacgtacaa aacgcgtgcg gaagtgaaat
     3781 ttgaaggcga taccctggta aaccgcattg agctgaaagg cattgacttt aaagaagacg
     3841 gcaatatcct gggccataag ctggaataca attttaacag ccacaatgtt tacatcaccg
     3901 ccgataaaca aaaaaatggc attaaagcga attttaaaat tcgccacaac gtggaggatg
     3961 gcagcgtgca gctggctgat cactaccagc aaaacactcc aatcggtgat ggtcctgttc
     4021 tgctgccaga caatcactat ctgagcacgc aaagcgttct gtctaaagat ccgaacgaga
     4081 aacgcgatca tatggttctg ctggagttcg taaccgcagc gggcatcacg catggtatgg
     4141 atgaactgta caaatgatga agatctccag gcatcaaata aaacgaaagg ctcagtcgaa
     4201 agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctctact agagtcacac
     4261 tggctcacct tcgggtgggc ctttctgcgt ttata
//
