LOCUS       sequence-276366-        2809 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Low phosphate inducible promoter phoEp for GFP expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2809)
  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 2809)
  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-phoEp-GFPuv
FEATURES             Location/Qualifiers
     source          1..2809
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(242..244,245..247,248..961)
                     /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(275..303)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     898..918
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      1148..1179
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        1180..1282
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1283..2098
                     /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(1356..1375)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2119..2146
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2158..2745)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2238..2257)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2767..2796)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 cagtccagtt acgctggagt ctgaggctcg tcctgaatga tatcaagctt gaattcgtta
       61 gcatggcgtt ttgttgcgcg ggatcagcaa gcctagcggc agttgtttac gcttttatta
      121 cagatttaat aaattaccac attttaagaa tattattaat ctgtaatata tctttaacaa
      181 tctcaggtta aaaactttcc tgttttcaac gggactctcc cgctgtgtag gaggataatc
      241 tatggctagc aaaggagaag aacttttcac tggagttgtc ccaattcttg ttgaattaga
      301 tggtgatgtt aatgggcaca aattttctgt cagtggagag ggtgaaggtg atgctacata
      361 cggaaagctt acccttaaat ttatttgcac tactggaaaa ctacctgttc catggccaac
      421 acttgtcact actttctctt atggtgttca atgcttttcc cgttatccgg atcatatgaa
      481 acggcatgac tttttcaaga gtgccatgcc cgaaggttat gtacaggaac gcactatatc
      541 tttcaaagat gacgggaact acaagacgcg tgctgaagtc aagtttgaag gtgataccct
      601 tgttaatcgt atcgagttaa aaggtattga ttttaaagaa gatggaaaca ttctcggaca
      661 caaactcgag tacaactata actcacacaa tgtatacatc acggcagaca aacaaaagaa
      721 tggaatcaaa gctaacttca aaattcgcca caacattgaa gatggatccg ttcaactagc
      781 agaccattat caacaaaata ctccaattgg cgatggccct gtccttttac cagacaacca
      841 ttacctgtcg acacaatctg ccctttcgaa agatcccaac gaaaagcgtg accacatggt
      901 ccttcttgag tttgtaactg ctgctgggat tacacatggc atggatgagc tctacaaata
      961 atgaggatcc ccggcttatc ggtcagtttc acctgattta cgtaaaaacc cgcttcggcg
     1021 ggtttttgct tttggagggg cagaaagatg aatgactgtc cacgacgcta tacccaaaag
     1081 aaagacgaat tctctagata tcgctcaata ctgaccattt aaatcatacc tgacctccat
     1141 agcagaaagt caaaagcctc cgaccggagg cttttgactt gatcggcacg taagaggttc
     1201 caactttcac cataatgaaa taagatcact accgggcgta ttttttgagt tatcgagatt
     1261 ttcaggagct aaggaagcta aaatgagcca tattcaacgg gaaacgtctt gctcgaggcc
     1321 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt
     1381 cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc cagagttgtt
     1441 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcaggctaaa
     1501 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
     1561 tgcatggtta ctcaccactg cgatcccagg gaaaacagca ttccaggtat tagaagaata
     1621 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc
     1681 gattcctgtt tgtaattgtc cttttaacgg cgatcgcgta tttcgtctcg ctcaggcgca
     1741 atcacgaatg aataacggtt tggttggtgc gagtgatttt gatgacgagc gtaatggctg
     1801 gcctgttgaa caagtctgga aagaaatgca taagcttttg ccattctcac cggattcagt
     1861 cgtcacccat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
     1921 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg
     1981 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat
     2041 tgataatcct gatatgaata aattgcagtt tcacttgatg ctcgatgagt ttttctgagg
     2101 gcccaaatgt aatcacctgg ctcaccttcg ggtgggcctt tctgcgttgc tggcgttttt
     2161 ccataggctc cgcccccctg acgagcatca caaaaatcga tgctcaagtc agaggtggcg
     2221 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     2281 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     2341 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     2401 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     2461 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     2521 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     2581 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctc
     2641 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     2701 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatt
     2761 ttctaccgaa gaaaggccca cccgtgaagg tgagccagtg agttgattg
//
