LOCUS       sequence-276369-        2745 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Low phosphate inducible promoter pstSp for GFP expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2745)
  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 2745)
  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-pstSp-GFPuv
FEATURES             Location/Qualifiers
     source          1..2745
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(178..180,181..183,184..897)
                     /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(211..239)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     834..854
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      1084..1115
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        1116..1218
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1219..2034
                     /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(1292..1311)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2055..2082
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2094..2681)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2174..2193)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2703..2732)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 cagtccagtt acgctggagt ctgaggctcg tcctgaatga tatcaagctt gaattcgtta
       61 agactttatc tctctgtcat aaaactgtca tattccttac atataactgt cacctgtttg
      121 tcctattttg cttctcgtag ccaacaaaca atgctttatg atgtaggagg ataatctatg
      181 gctagcaaag gagaagaact tttcactgga gttgtcccaa ttcttgttga attagatggt
      241 gatgttaatg ggcacaaatt ttctgtcagt ggagagggtg aaggtgatgc tacatacgga
      301 aagcttaccc ttaaatttat ttgcactact ggaaaactac ctgttccatg gccaacactt
      361 gtcactactt tctcttatgg tgttcaatgc ttttcccgtt atccggatca tatgaaacgg
      421 catgactttt tcaagagtgc catgcccgaa ggttatgtac aggaacgcac tatatctttc
      481 aaagatgacg ggaactacaa gacgcgtgct gaagtcaagt ttgaaggtga tacccttgtt
      541 aatcgtatcg agttaaaagg tattgatttt aaagaagatg gaaacattct cggacacaaa
      601 ctcgagtaca actataactc acacaatgta tacatcacgg cagacaaaca aaagaatgga
      661 atcaaagcta acttcaaaat tcgccacaac attgaagatg gatccgttca actagcagac
      721 cattatcaac aaaatactcc aattggcgat ggccctgtcc ttttaccaga caaccattac
      781 ctgtcgacac aatctgccct ttcgaaagat cccaacgaaa agcgtgacca catggtcctt
      841 cttgagtttg taactgctgc tgggattaca catggcatgg atgagctcta caaataatga
      901 ggatccccgg cttatcggtc agtttcacct gatttacgta aaaacccgct tcggcgggtt
      961 tttgcttttg gaggggcaga aagatgaatg actgtccacg acgctatacc caaaagaaag
     1021 acgaattctc tagatatcgc tcaatactga ccatttaaat catacctgac ctccatagca
     1081 gaaagtcaaa agcctccgac cggaggcttt tgacttgatc ggcacgtaag aggttccaac
     1141 tttcaccata atgaaataag atcactaccg ggcgtatttt ttgagttatc gagattttca
     1201 ggagctaagg aagctaaaat gagccatatt caacgggaaa cgtcttgctc gaggccgcga
     1261 ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg
     1321 caatcaggtg cgacaatcta tcgattgtat gggaagcccg atgcgccaga gttgtttctg
     1381 aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag gctaaactgg
     1441 ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca
     1501 tggttactca ccactgcgat cccagggaaa acagcattcc aggtattaga agaatatcct
     1561 gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt
     1621 cctgtttgta attgtccttt taacggcgat cgcgtatttc gtctcgctca ggcgcaatca
     1681 cgaatgaata acggtttggt tggtgcgagt gattttgatg acgagcgtaa tggctggcct
     1741 gttgaacaag tctggaaaga aatgcataag cttttgccat tctcaccgga ttcagtcgtc
     1801 acccatggtg atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt
     1861 attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac
     1921 tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat
     1981 aatcctgata tgaataaatt gcagtttcac ttgatgctcg atgagttttt ctgagggccc
     2041 aaatgtaatc acctggctca ccttcgggtg ggcctttctg cgttgctggc gtttttccat
     2101 aggctccgcc cccctgacga gcatcacaaa aatcgatgct caagtcagag gtggcgaaac
     2161 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2221 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2281 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2341 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2401 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     2461 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     2521 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt tacctcggaa
     2581 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     2641 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgattttct
     2701 accgaagaaa ggcccacccg tgaaggtgag ccagtgagtt gattg
//
