LOCUS       sequence-276370-        2738 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Low phosphate inducible promoter ugpBp for GFP expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2738)
  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 2738)
  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-ugpBp-GFPuv
FEATURES             Location/Qualifiers
     source          1..2738
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(171..173,174..176,177..890)
                     /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(204..232)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     827..847
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      1077..1108
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        1109..1211
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1212..2027
                     /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(1285..1304)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2048..2075
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2087..2674)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2167..2186)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2696..2725)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 cagtccagtt acgctggagt ctgaggctcg tcctgaatga tatcaagctt gaattcgttt
       61 ctttctgaca ccttactatc ttacaaatgt aacaaaaaag ttatttttct gtaattcgag
      121 catgtcatgt taccccgcga gcataaaacg cgtgtgtagg aggataatct atggctagca
      181 aaggagaaga acttttcact ggagttgtcc caattcttgt tgaattagat ggtgatgtta
      241 atgggcacaa attttctgtc agtggagagg gtgaaggtga tgctacatac ggaaagctta
      301 cccttaaatt tatttgcact actggaaaac tacctgttcc atggccaaca cttgtcacta
      361 ctttctctta tggtgttcaa tgcttttccc gttatccgga tcatatgaaa cggcatgact
      421 ttttcaagag tgccatgccc gaaggttatg tacaggaacg cactatatct ttcaaagatg
      481 acgggaacta caagacgcgt gctgaagtca agtttgaagg tgataccctt gttaatcgta
      541 tcgagttaaa aggtattgat tttaaagaag atggaaacat tctcggacac aaactcgagt
      601 acaactataa ctcacacaat gtatacatca cggcagacaa acaaaagaat ggaatcaaag
      661 ctaacttcaa aattcgccac aacattgaag atggatccgt tcaactagca gaccattatc
      721 aacaaaatac tccaattggc gatggccctg tccttttacc agacaaccat tacctgtcga
      781 cacaatctgc cctttcgaaa gatcccaacg aaaagcgtga ccacatggtc cttcttgagt
      841 ttgtaactgc tgctgggatt acacatggca tggatgagct ctacaaataa tgaggatccc
      901 cggcttatcg gtcagtttca cctgatttac gtaaaaaccc gcttcggcgg gtttttgctt
      961 ttggaggggc agaaagatga atgactgtcc acgacgctat acccaaaaga aagacgaatt
     1021 ctctagatat cgctcaatac tgaccattta aatcatacct gacctccata gcagaaagtc
     1081 aaaagcctcc gaccggaggc ttttgacttg atcggcacgt aagaggttcc aactttcacc
     1141 ataatgaaat aagatcacta ccgggcgtat tttttgagtt atcgagattt tcaggagcta
     1201 aggaagctaa aatgagccat attcaacggg aaacgtcttg ctcgaggccg cgattaaatt
     1261 ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag
     1321 gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg
     1381 gcaaaggtag cgttgccaat gatgttacag atgagatggt caggctaaac tggctgacgg
     1441 aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac
     1501 tcaccactgc gatcccaggg aaaacagcat tccaggtatt agaagaatat cctgattcag
     1561 gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt
     1621 gtaattgtcc ttttaacggc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga
     1681 ataacggttt ggttggtgcg agtgattttg atgacgagcg taatggctgg cctgttgaac
     1741 aagtctggaa agaaatgcat aagcttttgc cattctcacc ggattcagtc gtcacccatg
     1801 gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg
     1861 ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg
     1921 gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg
     1981 atatgaataa attgcagttt cacttgatgc tcgatgagtt tttctgaggg cccaaatgta
     2041 atcacctggc tcaccttcgg gtgggccttt ctgcgttgct ggcgtttttc cataggctcc
     2101 gcccccctga cgagcatcac aaaaatcgat gctcaagtca gaggtggcga aacccgacag
     2161 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     2221 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     2281 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     2341 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     2401 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     2461 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     2521 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttacctcg gaaaaagagt
     2581 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     2641 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgattt tctaccgaag
     2701 aaaggcccac ccgtgaaggt gagccagtga gttgattg
//
