LOCUS       sequence-493046-        3957 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the SRTF1 transcription factor in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3957)
  TITLE     Example of directed evolution in E. coli to develop novel 
            ligand-binding regulators: evolving a progesterone-responsive 
            transcription factor to bind cortisol
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3957)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pREG-SRTF1
FEATURES             Location/Qualifiers
     source          1..3957
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        37..86
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     protein_bind    complement(939..972)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     promoter        983..1054
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1055..1123,1124..1915)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1273..1292)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     protein_bind    complement(2049..2082)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             2531..2722
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     2879..2898
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2973..2990)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3144..3731)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3224..3243)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aggtacaatt acagccatcg tagaagacac tggcagctgt caccggatgt gctttccggt
       61 ctgatgagtc cgtgaggacg aaacagcctc tacaaataat tttgtttaag ggcccaagtt
      121 cacttaaaaa ggagatcaac aatgaaagca attttcgtac tgaaacatct taatcatgcg
      181 ccggtgtttt aaatatgagc tcgacggctg aacgtattcg tccgggacgc tctggtattt
      241 tggcagcggc cacccgcctg tttgcaactc acggggtatc tggtacgtcg ctgcagcaga
      301 ttgccgacgc aacgggcatt acaaaagccg ctgtgtacca ccacttccct acgaaagagg
      361 aggtcgtcgt cgccgttctt gcacctgctt tagaagccat tcagggcatt gtccgtaccg
      421 ctggtgcgca tgaggacccg cgtgctgcaa cagaggctgc aattatcgga cttgcggacc
      481 aggcagttac gcatcgtcaa cgctgggcag tccttttaca ggacgccgcc gttgaggagt
      541 atgtacgtaa taatcctgat cacgatgaat tatttacgcg ccttcgcctg cttctgacgg
      601 ggcccgatcc tacaccgggt acacgtttgc aggtttcgct gttcctgtct ggtttattgg
      661 gaccggcgca ggatccatca tgcgccgaca ttgatgatga tgcgttacgt gcaggcatcg
      721 tacgcgctgg acgccgcctt ctgttagcag acgacgatgc ctaatcctag tcttctcgcc
      781 actttcagcc aaaaaactta agaccgccgg tcttgtccac taccttgcag taatgcggtg
      841 gacaggatcg gcggttttct tttctcttct caacaccctt cgcgtcaaca cttttccgcc
      901 aaggagacgg ttggtcaggt tttcgggagg tgtggctgga agttcctata ctttctagag
      961 aataggaact tctttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     1021 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     1081 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     1141 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     1201 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     1261 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact
     1321 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     1381 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     1441 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     1501 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     1561 agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg
     1621 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     1681 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     1741 tgctgataaa tctggagccg gtgagcgtgg atcgcgcggt atcattgcag cactggggcc
     1801 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     1861 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaagttgt
     1921 gatggcggta ggaatgtaat cgttaatccg caaataacgt aaaaacccgc ttcggcgggt
     1981 ttttttatgg ggggagttta gggaaagagc atttgtcatc ccgttgaata tggctccctt
     2041 aacgtgagga agttcctata ctttctagag aataggaact tctacagatg gacttgggtt
     2101 ggcggcctag acgtgtgata gactcagtcg gcttcgatgt ttgagttgtg tgtttttaca
     2161 tcatatgaca tcaggagtct gcaaaacgtc tgcgacctga gcaacaacat gaatggtcat
     2221 cggtttccgt gtttcgtaaa gtctggaaac gcggaagtca gcgccctgca ccattatgtt
     2281 ccggatctgc atcgcaggat gctgctggct accctgtgga acacctacat ctgtattaac
     2341 gaagcgctgg cattgaccct gagtgatttt tctctggtcc cgccgcatcc ataccgccag
     2401 ttgtttaccc tcacaacgtt ccagtaaccg ggcatgttca tcatcagtaa cccgtatcgt
     2461 gagcatcctc tctcgtttca tcggtatcat tacccccatg aacagaaatc ccccttacac
     2521 ggaggcatca gtgaccaaac aggaaaaaac cgcccttaac atggcccgct ttatcagaag
     2581 ccagacatta acgcttctgg agaaactcaa cgagctggat gcggatgaac aggcagacat
     2641 ctgtgaatcg cttcacgacc acgctgatga gctttaccgc agctgcctcg cgcgtttcgg
     2701 tgatgacggt gaaaacctct gacacatgca gctcccgcag acggtcacag cttgtctgta
     2761 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     2821 gggcgcagcc atgacccagt cacgtagcga tagcggagtg tatactggct taactatgcg
     2881 gcatcagagc agattgtact gagagtgcac cggtgtgaaa taccgcacag atgcgtaagg
     2941 agaaaatacc gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     3001 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     3061 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     3121 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     3181 aattgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     3241 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     3301 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     3361 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     3421 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     3481 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     3541 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     3601 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     3661 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     3721 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     3781 actcacgtta aggccctctc caagaccgag ccatcaacaa agcgtctcgc tgaggtttca
     3841 tggagcctct ggttcatctc cggcaattaa aaaagcggct aaccacgccg ctttttttac
     3901 gtctgcagga acgggctgtc gacctttgaa aagttcgttt accgctagct cagtcct
//
