LOCUS       sequence-293811-        2850 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Low phosphate inducible promoter tktBp for GFP expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2850)
  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 2850)
  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-tktBp-GFPuv
FEATURES             Location/Qualifiers
     source          1..2850
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1..3,4..6,7..720)
                     /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(34..62)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     657..677
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      907..938
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        939..1041
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1042..1857
                     /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(1115..1134)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1878..1905
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(1917..2504)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1997..2016)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2526..2555)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 atggctagca aaggagaaga acttttcact ggagttgtcc caattcttgt tgaattagat
       61 ggtgatgtta atgggcacaa attttctgtc agtggagagg gtgaaggtga tgctacatac
      121 ggaaagctta cccttaaatt tatttgcact actggaaaac tacctgttcc atggccaaca
      181 cttgtcacta ctttctctta tggtgttcaa tgcttttccc gttatccgga tcatatgaaa
      241 cggcatgact ttttcaagag tgccatgccc gaaggttatg tacaggaacg cactatatct
      301 ttcaaagatg acgggaacta caagacgcgt gctgaagtca agtttgaagg tgataccctt
      361 gttaatcgta tcgagttaaa aggtattgat tttaaagaag atggaaacat tctcggacac
      421 aaactcgagt acaactataa ctcacacaat gtatacatca cggcagacaa acaaaagaat
      481 ggaatcaaag ctaacttcaa aattcgccac aacattgaag atggatccgt tcaactagca
      541 gaccattatc aacaaaatac tccaattggc gatggccctg tccttttacc agacaaccat
      601 tacctgtcga cacaatctgc cctttcgaaa gatcccaacg aaaagcgtga ccacatggtc
      661 cttcttgagt ttgtaactgc tgctgggatt acacatggca tggatgagct ctacaaataa
      721 tgaggatccc cggcttatcg gtcagtttca cctgatttac gtaaaaaccc gcttcggcgg
      781 gtttttgctt ttggaggggc agaaagatga atgactgtcc acgacgttat acccaaaaga
      841 aagacgaatt ctctagatat cgctcaatac tgaccattta aatcatacct gacctccata
      901 gcagaaagtc aaaagcctcc gaccggaggc ttttgacttg atcggcacgt aagaggttcc
      961 aactttcacc ataatgaaat aagatcacta ccgggcgtat tttttgagtt atcgagattt
     1021 tcaggagcta aggaagctaa aatgagccat attcaacggg aaacgtcttg ctcgaggccg
     1081 cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc
     1141 gggcaatcag gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc agagttgttt
     1201 ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt caggctaaac
     1261 tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat
     1321 gcatggttac tcaccactgc gatcccaggg aaaacagcat tccaggtatt agaagaatat
     1381 cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg
     1441 attcctgttt gtaattgtcc ttttaacggc gatcgcgtat ttcgtctcgc tcaggcgcaa
     1501 tcacgaatga ataacggttt ggttggtgcg agtgattttg atgacgagcg taatggctgg
     1561 cctgttgaac aagtctggaa agaaatgcat aagcttttgc cattctcacc ggattcagtc
     1621 gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt
     1681 tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg
     1741 aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt
     1801 gataatcctg atatgaataa attgcagttt cacttgatgc tcgatgagtt tttctgaggg
     1861 cccaaatgta atcacctggc tcaccttcgg gtgggccttt ctgcgttgct ggcgtttttc
     1921 cataggctcc gcccccctga cgagcatcac aaaaatcgat gctcaagtca gaggtggcga
     1981 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     2041 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     2101 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     2161 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     2221 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     2281 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     2341 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttacctcg
     2401 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     2461 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgattt
     2521 tctaccgaag aaaggcccac ccgtgaaggt gagccagtga gttgattgca gtccagttac
     2581 gctggagtct gaggctcgtc ctgaatgata tcaagcttga attcgttctg caggaaaaag
     2641 tttcgccagt ggtacgtaaa ttaatcccac cttctcagac gttcccacac ccagctccca
     2701 tgagcgaagc ggagttccgt tgggagcaca atcaggatgc gatggcggta gaaaaactgt
     2761 ctgaaggcat tcgtctgttc gccgttgatc aacgcaaact ggaagatctt cttgccgcca
     2821 aactataaac cagccacagg aggataatct
//
