LOCUS       sequence-276373-        2762 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Low phosphate inducible promoter ytfKp for GFP expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2762)
  AUTHORS   Moreb EA, Ye Z, Efromson JP, Hennigan JN, Menacho-Melgar R, Lynch MD
  TITLE     Media Robustness and Scalability of Phosphate Regulated Promoters 
            Useful for Two-Stage Autoinduction in E. coli.
  JOURNAL   ACS Synth Biol. 2020 Jun 19;9(6):1483-1486. doi: 
            10.1021/acssynbio.0c00182. Epub 2020 May 27.
  PUBMED    32353228
REFERENCE   2  (bases 1 to 2762)
  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.0c00182"; journalName: "ACS Synth Biol"; date: 
            "2020-06-19- 19"; volume: "9"; issue: "6"; pages: "1483-1486"
COMMENT     Sequence Label: pHCKan-ytfKp-GFPuv
FEATURES             Location/Qualifiers
     source          1..2762
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(195..197,198..200,201..914)
                     /codon_start=1
                     /product="Cycle 3 GFP (Crameri et al., 1996)"
                     /label=Cycle 3 GFP
                     /note="mammalian codon-optimized"
                     /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(228..256)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     851..871
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      1101..1132
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        1133..1235
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1236..2051
                     /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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVGA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1309..1328)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2072..2099
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2111..2698)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2191..2210)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2720..2749)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 cagtccagtt acgctggagt ctgaggctcg tcctgaatga tatcaagctt gaattcgttc
       61 ttcggtaaaa agataattct gaataattgt aacctttagg taaaaaaagt tatacgcggt
      121 ggaaacattg cccggatagt ctatagtcac taagcattaa aatttgcgcc tcataatatg
      181 taggaggata atctatggct agcaaaggag aagaactttt cactggagtt gtcccaattc
      241 ttgttgaatt agatggtgat gttaatgggc acaaattttc tgtcagtgga gagggtgaag
      301 gtgatgctac atacggaaag cttaccctta aatttatttg cactactgga aaactacctg
      361 ttccatggcc aacacttgtc actactttct cttatggtgt tcaatgcttt tcccgttatc
      421 cggatcatat gaaacggcat gactttttca agagtgccat gcccgaaggt tatgtacagg
      481 aacgcactat atctttcaaa gatgacggga actacaagac gcgtgctgaa gtcaagtttg
      541 aaggtgatac ccttgttaat cgtatcgagt taaaaggtat tgattttaaa gaagatggaa
      601 acattctcgg acacaaactc gagtacaact ataactcaca caatgtatac atcacggcag
      661 acaaacaaaa gaatggaatc aaagctaact tcaaaattcg ccacaacatt gaagatggat
      721 ccgttcaact agcagaccat tatcaacaaa atactccaat tggcgatggc cctgtccttt
      781 taccagacaa ccattacctg tcgacacaat ctgccctttc gaaagatccc aacgaaaagc
      841 gtgaccacat ggtccttctt gagtttgtaa ctgctgctgg gattacacat ggcatggatg
      901 agctctacaa ataatgagga tccccggctt atcggtcagt ttcacctgat ttacgtaaaa
      961 acccgcttcg gcgggttttt gcttttggag gggcagaaag atgaatgact gtccacgacg
     1021 ctatacccaa aagaaagacg aattctctag atatcgctca atactgacca tttaaatcat
     1081 acctgacctc catagcagaa agtcaaaagc ctccgaccgg aggcttttga cttgatcggc
     1141 acgtaagagg ttccaacttt caccataatg aaataagatc actaccgggc gtattttttg
     1201 agttatcgag attttcagga gctaaggaag ctaaaatgag ccatattcaa cgggaaacgt
     1261 cttgctcgag gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg
     1321 ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg attgtatggg aagcccgatg
     1381 cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga
     1441 tggtcaggct aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc
     1501 gtactcctga tgatgcatgg ttactcacca ctgcgatccc agggaaaaca gcattccagg
     1561 tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc
     1621 gccggttgca ttcgattcct gtttgtaatt gtccttttaa cggcgatcgc gtatttcgtc
     1681 tcgctcaggc gcaatcacga atgaataacg gtttggttgg tgcgagtgat tttgatgacg
     1741 agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataagctt ttgccattct
     1801 caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg
     1861 ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc
     1921 ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc
     1981 aaaaatatgg tattgataat cctgatatga ataaattgca gtttcacttg atgctcgatg
     2041 agtttttctg agggcccaaa tgtaatcacc tggctcacct tcgggtgggc ctttctgcgt
     2101 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgatgctcaa
     2161 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     2221 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     2281 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     2341 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     2401 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     2461 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     2521 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     2581 agccagttac ctcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     2641 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     2701 agatcctttg attttctacc gaagaaaggc ccacccgtga aggtgagcca gtgagttgat
     2761 tg
//
