LOCUS       sequence-276374-        2779 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Low phosphate inducible promoter phoUp for GFP expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2779)
  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 2779)
  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-phoUp-GFPuv
FEATURES             Location/Qualifiers
     source          1..2779
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(212..214,215..217,218..931)
                     /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(245..273)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     868..888
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      1118..1149
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        1150..1252
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1253..2068
                     /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(1326..1345)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2089..2116
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2128..2715)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2208..2227)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2737..2766)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 cagtccagtt acgctggagt ctgaggctcg tcctgaatga tatcaagctt gaattcgtta
       61 ccgaactgaa gcaggattac accgtggtga tcgtcaccca caacatgcag caggctgcgc
      121 gttgttccga ccacacggcg tttatgtacc tgggcgaatt gattgagttc agcaacacgg
      181 acgatctgtt caccatgtag gaggataatc tatggctagc aaaggagaag aacttttcac
      241 tggagttgtc ccaattcttg ttgaattaga tggtgatgtt aatgggcaca aattttctgt
      301 cagtggagag ggtgaaggtg atgctacata cggaaagctt acccttaaat ttatttgcac
      361 tactggaaaa ctacctgttc catggccaac acttgtcact actttctctt atggtgttca
      421 atgcttttcc cgttatccgg atcatatgaa acggcatgac tttttcaaga gtgccatgcc
      481 cgaaggttat gtacaggaac gcactatatc tttcaaagat gacgggaact acaagacgcg
      541 tgctgaagtc aagtttgaag gtgataccct tgttaatcgt atcgagttaa aaggtattga
      601 ttttaaagaa gatggaaaca ttctcggaca caaactcgag tacaactata actcacacaa
      661 tgtatacatc acggcagaca aacaaaagaa tggaatcaaa gctaacttca aaattcgcca
      721 caacattgaa gatggatccg ttcaactagc agaccattat caacaaaata ctccaattgg
      781 cgatggccct gtccttttac cagacaacca ttacctgtcg acacaatctg ccctttcgaa
      841 agatcccaac gaaaagcgtg accacatggt ccttcttgag tttgtaactg ctgctgggat
      901 tacacatggc atggatgagc tctacaaata atgaggatcc ccggcttatc ggtcagtttc
      961 acctgattta cgtaaaaacc cgcttcggcg ggtttttgct tttggagggg cagaaagatg
     1021 aatgactgtc cacgacgcta tacccaaaag aaagacgaat tctctagata tcgctcaata
     1081 ctgaccattt aaatcatacc tgacctccat agcagaaagt caaaagcctc cgaccggagg
     1141 cttttgactt gatcggcacg taagaggttc caactttcac cataatgaaa taagatcact
     1201 accgggcgta ttttttgagt tatcgagatt ttcaggagct aaggaagcta aaatgagcca
     1261 tattcaacgg gaaacgtctt gctcgaggcc gcgattaaat tccaacatgg atgctgattt
     1321 atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt
     1381 gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa
     1441 tgatgttaca gatgagatgg tcaggctaaa ctggctgacg gaatttatgc ctcttccgac
     1501 catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatcccagg
     1561 gaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc
     1621 gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacgg
     1681 cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttggtgc
     1741 gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca
     1801 taagcttttg ccattctcac cggattcagt cgtcacccat ggtgatttct cacttgataa
     1861 ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc
     1921 agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt
     1981 acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt
     2041 tcacttgatg ctcgatgagt ttttctgagg gcccaaatgt aatcacctgg ctcaccttcg
     2101 ggtgggcctt tctgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     2161 caaaaatcga tgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2221 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2281 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     2341 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     2401 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     2461 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     2521 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     2581 tatctgcgct ctgctgaagc cagttacctc ggaaaaagag ttggtagctc ttgatccggc
     2641 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     2701 aaaaaaggat ctcaagaaga tcctttgatt ttctaccgaa gaaaggccca cccgtgaagg
     2761 tgagccagtg agttgattg
//
