LOCUS       sequence-276364-        2748 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Low phosphate inducible promoter phoBp for GFP expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2748)
  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 2748)
  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-phoBp-GFPuv
FEATURES             Location/Qualifiers
     source          1..2748
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(181..183,184..186,187..900)
                     /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(214..242)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     837..857
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      1087..1118
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        1119..1221
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1222..2037
                     /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(1295..1314)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2058..2085
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2097..2684)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2177..2196)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2706..2735)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 cagtccagtt acgctggagt ctgaggctcg tcctgaatga tatcaagctt gaattcgttg
       61 ccacggaaat caataacctg aagatatgtg cgacgagctt ttcataaatc tgtcataaat
      121 ctgacgcata atgacgtcgc attaatgatc gcaacctatt tatttgtagg aggataatct
      181 atggctagca aaggagaaga acttttcact ggagttgtcc caattcttgt tgaattagat
      241 ggtgatgtta atgggcacaa attttctgtc agtggagagg gtgaaggtga tgctacatac
      301 ggaaagctta cccttaaatt tatttgcact actggaaaac tacctgttcc atggccaaca
      361 cttgtcacta ctttctctta tggtgttcaa tgcttttccc gttatccgga tcatatgaaa
      421 cggcatgact ttttcaagag tgccatgccc gaaggttatg tacaggaacg cactatatct
      481 ttcaaagatg acgggaacta caagacgcgt gctgaagtca agtttgaagg tgataccctt
      541 gttaatcgta tcgagttaaa aggtattgat tttaaagaag atggaaacat tctcggacac
      601 aaactcgagt acaactataa ctcacacaat gtatacatca cggcagacaa acaaaagaat
      661 ggaatcaaag ctaacttcaa aattcgccac aacattgaag atggatccgt tcaactagca
      721 gaccattatc aacaaaatac tccaattggc gatggccctg tccttttacc agacaaccat
      781 tacctgtcga cacaatctgc cctttcgaaa gatcccaacg aaaagcgtga ccacatggtc
      841 cttcttgagt ttgtaactgc tgctgggatt acacatggca tggatgagct ctacaaataa
      901 tgaggatccc cggcttatcg gtcagtttca cctgatttac gtaaaaaccc gcttcggcgg
      961 gtttttgctt ttggaggggc agaaagatga atgactgtcc acgacgctat acccaaaaga
     1021 aagacgaatt ctctagatat cgctcaatac tgaccattta aatcatacct gacctccata
     1081 gcagaaagtc aaaagcctcc gaccggaggc ttttgacttg atcggcacgt aagaggttcc
     1141 aactttcacc ataatgaaat aagatcacta ccgggcgtat tttttgagtt atcgagattt
     1201 tcaggagcta aggaagctaa aatgagccat attcaacggg aaacgtcttg ctcgaggccg
     1261 cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc
     1321 gggcaatcag gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc agagttgttt
     1381 ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt caggctaaac
     1441 tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat
     1501 gcatggttac tcaccactgc gatcccaggg aaaacagcat tccaggtatt agaagaatat
     1561 cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg
     1621 attcctgttt gtaattgtcc ttttaacggc gatcgcgtat ttcgtctcgc tcaggcgcaa
     1681 tcacgaatga ataacggttt ggttggtgcg agtgattttg atgacgagcg taatggctgg
     1741 cctgttgaac aagtctggaa agaaatgcat aagcttttgc cattctcacc ggattcagtc
     1801 gtcacccatg gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt
     1861 tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg
     1921 aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt
     1981 gataatcctg atatgaataa attgcagttt cacttgatgc tcgatgagtt tttctgaggg
     2041 cccaaatgta atcacctggc tcaccttcgg gtgggccttt ctgcgttgct ggcgtttttc
     2101 cataggctcc gcccccctga cgagcatcac aaaaatcgat gctcaagtca gaggtggcga
     2161 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     2221 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     2281 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     2341 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     2401 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     2461 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     2521 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttacctcg
     2581 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     2641 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgattt
     2701 tctaccgaag aaaggcccac ccgtgaaggt gagccagtga gttgattg
//
