LOCUS       sequence-276317-        2747 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Low phosphate inducible promoter amnp  for GFP expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2747)
  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 2747)
  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-amnp-GFPuv
FEATURES             Location/Qualifiers
     source          1..2747
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(180..182,183..185,186..899)
                     /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(213..241)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     836..856
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      1086..1117
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        1118..1220
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1221..2036
                     /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(1294..1313)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2057..2084
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2096..2683)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2176..2195)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2705..2734)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 cagtccagtt acgctggagt ctgaggctcg tcctgaatga tatcaagctt gaattcgtta
       61 gacagtcaac gcgcttgata gcctggcgaa gatcatccga tcttcgcctt acacttttgt
      121 ttcacatttc tgtgacatac tatcggatgt gcggtaattg tattgtagga ggataatcta
      181 tggctagcaa aggagaagaa cttttcactg gagttgtccc aattcttgtt gaattagatg
      241 gtgatgttaa tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat gctacatacg
      301 gaaagcttac ccttaaattt atttgcacta ctggaaaact acctgttcca tggccaacac
      361 ttgtcactac tttctcttat ggtgttcaat gcttttcccg ttatccggat catatgaaac
      421 ggcatgactt tttcaagagt gccatgcccg aaggttatgt acaggaacgc actatatctt
      481 tcaaagatga cgggaactac aagacgcgtg ctgaagtcaa gtttgaaggt gatacccttg
      541 ttaatcgtat cgagttaaaa ggtattgatt ttaaagaaga tggaaacatt ctcggacaca
      601 aactcgagta caactataac tcacacaatg tatacatcac ggcagacaaa caaaagaatg
      661 gaatcaaagc taacttcaaa attcgccaca acattgaaga tggatccgtt caactagcag
      721 accattatca acaaaatact ccaattggcg atggccctgt ccttttacca gacaaccatt
      781 acctgtcgac acaatctgcc ctttcgaaag atcccaacga aaagcgtgac cacatggtcc
      841 ttcttgagtt tgtaactgct gctgggatta cacatggcat ggatgagctc tacaaataat
      901 gaggatcccc ggcttatcgg tcagtttcac ctgatttacg taaaaacccg cttcggcggg
      961 tttttgcttt tggaggggca gaaagatgaa tgactgtcca cgacgctata cccaaaagaa
     1021 agacgaattc tctagatatc gctcaatact gaccatttaa atcatacctg acctccatag
     1081 cagaaagtca aaagcctccg accggaggct tttgacttga tcggcacgta agaggttcca
     1141 actttcacca taatgaaata agatcactac cgggcgtatt ttttgagtta tcgagatttt
     1201 caggagctaa ggaagctaaa atgagccata ttcaacggga aacgtcttgc tcgaggccgc
     1261 gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg
     1321 ggcaatcagg tgcgacaatc tatcgattgt atgggaagcc cgatgcgcca gagttgtttc
     1381 tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc aggctaaact
     1441 ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg
     1501 catggttact caccactgcg atcccaggga aaacagcatt ccaggtatta gaagaatatc
     1561 ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga
     1621 ttcctgtttg taattgtcct tttaacggcg atcgcgtatt tcgtctcgct caggcgcaat
     1681 cacgaatgaa taacggtttg gttggtgcga gtgattttga tgacgagcgt aatggctggc
     1741 ctgttgaaca agtctggaaa gaaatgcata agcttttgcc attctcaccg gattcagtcg
     1801 tcacccatgg tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt
     1861 gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga
     1921 actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg
     1981 ataatcctga tatgaataaa ttgcagtttc acttgatgct cgatgagttt ttctgagggc
     2041 ccaaatgtaa tcacctggct caccttcggg tgggcctttc tgcgttgctg gcgtttttcc
     2101 ataggctccg cccccctgac gagcatcaca aaaatcgatg ctcaagtcag aggtggcgaa
     2161 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     2221 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     2281 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     2341 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     2401 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     2461 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     2521 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttacctcgg
     2581 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     2641 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatttt
     2701 ctaccgaaga aaggcccacc cgtgaaggtg agccagtgag ttgattg
//
