LOCUS       sequence-276361-        2829 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Low phosphate inducible promoter mipAp for GFP expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2829)
  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 2829)
  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-mipAp-GFPuv
FEATURES             Location/Qualifiers
     source          1..2829
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(262..264,265..267,268..981)
                     /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(295..323)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     918..938
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      1168..1199
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        1200..1302
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1303..2118
                     /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(1376..1395)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2139..2166
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2178..2765)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2258..2277)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2787..2816)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 cagtccagtt acgctggagt ctgaggctcg tcctgaatga tatcaagctt gaattcgttc
       61 atccataaat tttgcataat taatgtaaag accaggctcg ccagtaacgc taaattcatt
      121 tggctgtaag cgcggtgtca tccgcgtcag gaaaattaaa cagttacttt aaaaaatgaa
      181 aacgtaaaaa ggttgggttt cgatgtattg acgggtaaac tttgtcgccc gctaaacatt
      241 tgttttgtag gaggataatc tatggctagc aaaggagaag aacttttcac tggagttgtc
      301 ccaattcttg ttgaattaga tggtgatgtt aatgggcaca aattttctgt cagtggagag
      361 ggtgaaggtg atgctacata cggaaagctt acccttaaat ttatttgcac tactggaaaa
      421 ctacctgttc catggccaac acttgtcact actttctctt atggtgttca atgcttttcc
      481 cgttatccgg atcatatgaa acggcatgac tttttcaaga gtgccatgcc cgaaggttat
      541 gtacaggaac gcactatatc tttcaaagat gacgggaact acaagacgcg tgctgaagtc
      601 aagtttgaag gtgataccct tgttaatcgt atcgagttaa aaggtattga ttttaaagaa
      661 gatggaaaca ttctcggaca caaactcgag tacaactata actcacacaa tgtatacatc
      721 acggcagaca aacaaaagaa tggaatcaaa gctaacttca aaattcgcca caacattgaa
      781 gatggatccg ttcaactagc agaccattat caacaaaata ctccaattgg cgatggccct
      841 gtccttttac cagacaacca ttacctgtcg acacaatctg ccctttcgaa agatcccaac
      901 gaaaagcgtg accacatggt ccttcttgag tttgtaactg ctgctgggat tacacatggc
      961 atggatgagc tctacaaata atgaggatcc ccggcttatc ggtcagtttc acctgattta
     1021 cgtaaaaacc cgcttcggcg ggtttttgct tttggagggg cagaaagatg aatgactgtc
     1081 cacgacgcta tacccaaaag aaagacgaat tctctagata tcgctcaata ctgaccattt
     1141 aaatcatacc tgacctccat agcagaaagt caaaagcctc cgaccggagg cttttgactt
     1201 gatcggcacg taagaggttc caactttcac cataatgaaa taagatcact accgggcgta
     1261 ttttttgagt tatcgagatt ttcaggagct aaggaagcta aaatgagcca tattcaacgg
     1321 gaaacgtctt gctcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat
     1381 aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag
     1441 cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca
     1501 gatgagatgg tcaggctaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat
     1561 tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatcccagg gaaaacagca
     1621 ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg
     1681 ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacgg cgatcgcgta
     1741 tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttggtgc gagtgatttt
     1801 gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca taagcttttg
     1861 ccattctcac cggattcagt cgtcacccat ggtgatttct cacttgataa ccttattttt
     1921 gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac
     1981 caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg
     2041 ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt tcacttgatg
     2101 ctcgatgagt ttttctgagg gcccaaatgt aatcacctgg ctcaccttcg ggtgggcctt
     2161 tctgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     2221 tgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     2281 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     2341 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     2401 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     2461 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     2521 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     2581 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     2641 ctgctgaagc cagttacctc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     2701 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     2761 ctcaagaaga tcctttgatt ttctaccgaa gaaaggccca cccgtgaagg tgagccagtg
     2821 agttgattg
//
