LOCUS       sequence-276372-        2740 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Low phosphate inducible promoter yegHp for GFP expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2740)
  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 2740)
  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-yegHp-GFPuv
FEATURES             Location/Qualifiers
     source          1..2740
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(173..175,176..178,179..892)
                     /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(206..234)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     829..849
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      1079..1110
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        1111..1213
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1214..2029
                     /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(1287..1306)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2050..2077
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2089..2676)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2169..2188)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2698..2727)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 cagtccagtt acgctggagt ctgaggctcg tcctgaatga tatcaagctt gaattcgttg
       61 ttattaccct caaataatat gagataaata gtgctgcaac attgcatttt tgccccgatt
      121 tatccatgat cgaattgtga catttgtcat acaacgtgta ggaggataat ctatggctag
      181 caaaggagaa gaacttttca ctggagttgt cccaattctt gttgaattag atggtgatgt
      241 taatgggcac aaattttctg tcagtggaga gggtgaaggt gatgctacat acggaaagct
      301 tacccttaaa tttatttgca ctactggaaa actacctgtt ccatggccaa cacttgtcac
      361 tactttctct tatggtgttc aatgcttttc ccgttatccg gatcatatga aacggcatga
      421 ctttttcaag agtgccatgc ccgaaggtta tgtacaggaa cgcactatat ctttcaaaga
      481 tgacgggaac tacaagacgc gtgctgaagt caagtttgaa ggtgataccc ttgttaatcg
      541 tatcgagtta aaaggtattg attttaaaga agatggaaac attctcggac acaaactcga
      601 gtacaactat aactcacaca atgtatacat cacggcagac aaacaaaaga atggaatcaa
      661 agctaacttc aaaattcgcc acaacattga agatggatcc gttcaactag cagaccatta
      721 tcaacaaaat actccaattg gcgatggccc tgtcctttta ccagacaacc attacctgtc
      781 gacacaatct gccctttcga aagatcccaa cgaaaagcgt gaccacatgg tccttcttga
      841 gtttgtaact gctgctggga ttacacatgg catggatgag ctctacaaat aatgaggatc
      901 cccggcttat cggtcagttt cacctgattt acgtaaaaac ccgcttcggc gggtttttgc
      961 ttttggaggg gcagaaagat gaatgactgt ccacgacgct atacccaaaa gaaagacgaa
     1021 ttctctagat atcgctcaat actgaccatt taaatcatac ctgacctcca tagcagaaag
     1081 tcaaaagcct ccgaccggag gcttttgact tgatcggcac gtaagaggtt ccaactttca
     1141 ccataatgaa ataagatcac taccgggcgt attttttgag ttatcgagat tttcaggagc
     1201 taaggaagct aaaatgagcc atattcaacg ggaaacgtct tgctcgaggc cgcgattaaa
     1261 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     1321 aggtgcgaca atctatcgat tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     1381 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcaggctaa actggctgac
     1441 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     1501 actcaccact gcgatcccag ggaaaacagc attccaggta ttagaagaat atcctgattc
     1561 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     1621 ttgtaattgt ccttttaacg gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat
     1681 gaataacggt ttggttggtg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     1741 acaagtctgg aaagaaatgc ataagctttt gccattctca ccggattcag tcgtcactca
     1801 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     1861 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     1921 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     1981 tgatatgaat aaattgcagt ttcacttgat gctcgatgag tttttctgag ggcccaaatg
     2041 taatcacctg gctcaccttc gggtgggcct ttctgcgttg ctggcgtttt tccataggct
     2101 ccgcccccct gacgagcatc acaaaaatcg atgctcaagt cagaggtggc gaaacccgac
     2161 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     2221 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     2281 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     2341 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     2401 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     2461 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     2521 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct cggaaaaaga
     2581 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     2641 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat tttctaccga
     2701 agaaaggccc acccgtgaag gtgagccagt gagttgattg
//
