LOCUS       sequence-276363-        2768 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Low phosphate inducible promoter phoAp for GFP expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2768)
  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 2768)
  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-phoAp-GFPuv
FEATURES             Location/Qualifiers
     source          1..2768
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(201..203,204..206,207..920)
                     /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(234..262)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     857..877
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      1107..1138
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        1139..1241
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1242..2057
                     /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(1315..1334)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2078..2105
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2117..2704)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2197..2216)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2726..2755)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 cagtccagtt acgctggagt ctgaggctcg tcctgaatga tatcaagctt gaattcgttc
       61 gattacgtaa agaagttatt gaagcatcct cgtcagtaaa aagttaatct tttcaacagc
      121 tgtcataaag ttgtcacggc cgagacttat agtcgctttg tttttatttt ttaatgtatt
      181 tgtatgtagg aggataatct atggctagca aaggagaaga acttttcact ggagttgtcc
      241 caattcttgt tgaattagat ggtgatgtta atgggcacaa attttctgtc agtggagagg
      301 gtgaaggtga tgctacatac ggaaagctta cccttaaatt tatttgcact actggaaaac
      361 tacctgttcc atggccaaca cttgtcacta ctttctctta tggtgttcaa tgcttttccc
      421 gttatccgga tcatatgaaa cggcatgact ttttcaagag tgccatgccc gaaggttatg
      481 tacaggaacg cactatatct ttcaaagatg acgggaacta caagacgcgt gctgaagtca
      541 agtttgaagg tgataccctt gttaatcgta tcgagttaaa aggtattgat tttaaagaag
      601 atggaaacat tctcggacac aaactcgagt acaactataa ctcacacaat gtatacatca
      661 cggcagacaa acaaaagaat ggaatcaaag ctaacttcaa aattcgccac aacattgaag
      721 atggatccgt tcaactagca gaccattatc aacaaaatac tccaattggc gatggccctg
      781 tccttttacc agacaaccat tacctgtcga cacaatctgc cctttcgaaa gatcccaacg
      841 aaaagcgtga ccacatggtc cttcttgagt ttgtaactgc tgctgggatt acacatggca
      901 tggatgagct ctacaaataa tgaggatccc cggcttatcg gtcagtttca cctgatttac
      961 gtaaaaaccc gcttcggcgg gtttttgctt ttggaggggc agaaagatga atgactgtcc
     1021 acgacgctat acccaaaaga aagacgaatt ctctagatat cgctcaatac tgaccattta
     1081 aatcatacct gacctccata gcagaaagtc aaaagcctcc gaccggaggc ttttgacttg
     1141 atcggcacgt aagaggttcc aactttcacc ataatgaaat aagatcacta ccgggcgtat
     1201 tttttgagtt atcgagattt tcaggagcta aggaagctaa aatgagccat attcaacggg
     1261 aaacgtcttg ctcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata
     1321 aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgattg tatgggaagc
     1381 ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag
     1441 atgagatggt caggctaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt
     1501 ttatccgtac tcctgatgat gcatggttac tcaccactgc gatcccaggg aaaacagcat
     1561 tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt
     1621 tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacggc gatcgcgtat
     1681 ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttggtgcg agtgattttg
     1741 atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat aagcttttgc
     1801 cattctcacc ggattcagtc gtcacccatg gtgatttctc acttgataac cttatttttg
     1861 acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc
     1921 aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc
     1981 tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt cacttgatgc
     2041 tcgatgagtt tttctgaggg cccaaatgta atcacctggc tcaccttcgg gtgggccttt
     2101 ctgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgat
     2161 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     2221 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     2281 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     2341 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     2401 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     2461 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     2521 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     2581 tgctgaagcc agttacctcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     2641 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     2701 tcaagaagat cctttgattt tctaccgaag aaaggcccac ccgtgaaggt gagccagtga
     2761 gttgattg
//
