LOCUS       sequence-456070-        2817 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PUlaA promoter driving expression of sfgfp.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2817)
  AUTHORS   Piorino F, Styczynski MP
  TITLE     Harnessing Escherichia coli's Native Machinery for Detection of 
            Vitamin C (Ascorbate) Deficiency.
  JOURNAL   ACS Synth Biol. 2022 Nov 18;11(11):3592-3600. doi: 
            10.1021/acssynbio.2c00335. Epub 2022 Oct 27.
  PUBMED    36300901
REFERENCE   2  (bases 1 to 2817)
  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/acssynbio.2c00335"; journalName: "ACS Synth Biol"; date: 
            "2022-11-18- 18"; volume: "11"; issue: "11"; pages: "3592-3600"
COMMENT     Sequence Label: pCF
FEATURES             Location/Qualifiers
     source          1..2817
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      17..605
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     506..525
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     RBS             1014..1036
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1692..1739
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
     CDS             1740..1763
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1818..1836)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1832..1879
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1897..2712)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2620..2639
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2704..2723
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
       61 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
      121 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
      181 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
      241 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
      301 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      361 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      421 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      481 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      541 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      601 ggaaaaacgc cagcaacgcg atcccgcgaa atcgccattt accttcatga tagttcaatt
      661 cgaatcaata tgtgattggt tttgattaat cctgacacta ttttttcagg aaggcaatga
      721 ccattttttg actttttgcc agggaagttg ttgttgattt tgagtatgga aagatttaat
      781 ggaatgtgta attcaattaa ctaaatgaat ttaaatggat aattgtttcg ttgtgtgaat
      841 cccactctat ccatgtggaa ttatttgcgg gtcgcgtcac atttaatcat aaataatctt
      901 gttgtgatta cttttgaaaa ttagagtgag tgcacaacat tccgggtgtg tggaataccc
      961 ggttacctct tcttcaggag atcgttccac aacggtttcc ctctagaaat aattttgttt
     1021 aactttaaga aggagatata catatgagca aaggtgaaga actgtttacc ggcgttgtgc
     1081 cgattctggt ggaactggat ggcgatgtga acggtcacaa attcagcgtg cgtggtgaag
     1141 gtgaaggcga tgccacgatt ggcaaactga cgctgaaatt tatctgcacc accggcaaac
     1201 tgccggtgcc gtggccgacg ctggtgacca ccctgaccta tggcgttcag tgttttagtc
     1261 gctatccgga tcacatgaaa cgtcacgatt tctttaaatc tgcaatgccg gaaggctatg
     1321 tgcaggaacg tacgattagc tttaaagatg atggcaaata taaaacgcgc gccgttgtga
     1381 aatttgaagg cgataccctg gtgaaccgca ttgaactgaa aggcacggat tttaaagaag
     1441 atggcaatat cctgggccat aaactggaat acaactttaa tagccataat gtttatatta
     1501 cggcggataa acagaaaaat ggcatcaaag cgaattttac cgttcgccat aacgttgaag
     1561 atggcagtgt gcagctggca gatcattatc agcagaatac cccgattggt gatggtccgg
     1621 tgctgctgcc ggataatcat tatctgagca cgcagaccgt tctgtctaaa gatccgaacg
     1681 aaaaaggcac gcgggaccac atggttctgc acgaatatgt gaatgcggca ggtattacgt
     1741 ggagccatcc gcagttcgaa aaataagtcg accggctgct aacaaagccc gaaaggaagc
     1801 tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg
     1861 ggtcttgagg ggttttttgc tgaaagccaa ttctgattag aaaaactcat cgagcatcaa
     1921 atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt
     1981 ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg
     2041 gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat
     2101 aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag
     2161 cttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc
     2221 actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg
     2281 atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc
     2341 cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt
     2401 tttcccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt
     2461 gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac
     2521 atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc
     2581 atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc
     2641 atataaatca gcatccatgt tggaatttaa tcgcggcttc gagcaagacg tttcccgttg
     2701 aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt ttattgttca
     2761 tgatgatata tttttatctt gtgcaatgta acatcagaga ttttgagaca caacgtg
//
