LOCUS       sequence-399172-        3077 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses redox sensitive GFP (roGFP) after phosphate depletion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3077)
  AUTHORS   Hennigan JN, Menacho-Melgar R, Sarkar P, Golovsky M, Lynch MD
  TITLE     
  JOURNAL   Metab Eng. 2024 Sep;85:116-130. doi: 10.1016/j.ymben.2024.07.012. 
            Epub 2024 Jul 24.
  PUBMED    39059674
REFERENCE   2  (bases 1 to 3077)
  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"; journalName: "Metab 
            Eng."; date: "2024-09"; volume: "85:116-130. doi"; pages: " 
            10.1016/j.ymben.2024.07.012. Epub 2024 Jul 24"
COMMENT     Sequence Label: pSMART-HCKan-yibDp-roGFP2
FEATURES             Location/Qualifiers
     source          1..3077
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     8..27
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        8..26
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      205..236
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        237..339
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             340..1155
                     /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(413..432)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1176..1203
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(1215..1802)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1295..1314)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1824..1853)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     terminator      1937..2008
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2024..2051
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             join(2355..2357,2358..2360,2361..3074)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFISTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNCHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTCSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2400..2421)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2661..2680)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3008..3029
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 gctcttctaa tacgactcac tatagggccg gcttatcggt cagtttcacc tgatttacgt
       61 aaaaacccgc ttcggcgggt ttttgctttt ggaggggcag aaagatgaat gactgtccac
      121 gacgctatac ccaaaagaaa gacgaattct ctagatatcg ctcaatactg accatttaaa
      181 tcatacctga cctccatagc agaaagtcaa aagcctccga ccggaggctt ttgacttgat
      241 cggcacgtaa gaggttccaa ctttcaccat aatgaaataa gatcactacc gggcgtattt
      301 tttgagttat cgagattttc aggagctaag gaagctaaaa tgagccatat tcaacgggaa
      361 acgtcttgct cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa
      421 tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgattgta tgggaagccc
      481 gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat
      541 gagatggtca ggctaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt
      601 atccgtactc ctgatgatgc atggttactc accactgcga tcccagggaa aacagcattc
      661 caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc
      721 ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacggcga tcgcgtattt
      781 cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttggtgcgag tgattttgat
      841 gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa gcttttgcca
      901 ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac
      961 gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag
     1021 gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt
     1081 tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca cttgatgctc
     1141 gatgagtttt tctgagggcc caaatgtaat cacctggctc accttcgggt gggcctttct
     1201 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgatgc
     1261 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     1321 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     1381 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     1441 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     1501 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     1561 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     1621 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     1681 ctgaagccag ttacctcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     1741 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     1801 aagaagatcc tttgattttc taccgaagaa aggcccacca gtgaaggtga gccagtgagt
     1861 tgattgcagt ccagttacgc tggagtctga ggctcgtcct gaatgatatc aagcttgaat
     1921 tcgtttgccc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     1981 tatctgttgt ttgtcggtga acgctctcta ctagagtcac actggctcac cttcgggtgg
     2041 gcctttctgc gtttatacac agctaacacc acgtcgtccc tatctgctgc cctaggtcta
     2101 tgagtggttg ctggataacg tgcgtaattg tgctgatctc ttatatagct gctctcatta
     2161 tctctctacc ctgaagtgac tctctcacct gtaaaaataa tatctcacag gcttaatagt
     2221 ttcttaatac aaagcctgta aaacgtcagg ataacttcta tattcaggga gaccacaacg
     2281 gtttccctct acaaataatt ttgtttaact ttcgtagaag agcacttcac acttctggaa
     2341 aaaggagata taccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     2401 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     2461 gcgatgccac ctacggcaag ctgaccctga agttcatctc caccaccggc aagctgcccg
     2521 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
     2581 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     2641 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     2701 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     2761 acatcctggg gcacaagctg gagtacaact acaactgcca caacgtctat atcatggccg
     2821 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
     2881 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     2941 tgcccgacaa ccactacctg agcacctgct ccgccctgag caaagacccc aacgagaagc
     3001 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     3061 agctgtacaa gtaatga
//
