LOCUS       sequence-350152-        2859 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  QUAS directly upstream of a T7 promoter. Expresses GFP in BL21(DE3) 
            E. coli. Contains ColE1 origin of replication..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2859)
  AUTHORS   MacDonald IC, Seamons TR, Emmons JC, Javdan SB, Deans TL
  TITLE     Enhanced regulation of prokaryotic gene expression by a eukaryotic 
            transcriptional activator.
  JOURNAL   Nat Commun. 2021 Jul 5;12(1):4109. doi: 10.1038/s41467-021-24434-9.
  PUBMED    34226549
REFERENCE   2  (bases 1 to 2859)
  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: "Nat 
            Commun."; date: "2021-07-5"; volume: "12(1):4109. doi"; pages: " 
            10.1038/s41467-021-24434-9"
COMMENT     Sequence Label: QUAS-0-T7-GFP_ColE1
FEATURES             Location/Qualifiers
     source          1..2859
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(17..808,809..877))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     640..659
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(878..982)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1026..1044
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1062..1082
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             join(1091..1093,1094..1096,1097..1810)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGDEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLT
                     LKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKD
                     DGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGI
                     KVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVL
                     LEFVTAAGITLGMDELYK"
     primer_bind     complement(1139..1160)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1400..1419)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1747..1768
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      1883..1930
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      complement(2094..2682)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2174..2193)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(join(2770..2859,1..5))
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
ORIGIN
        1 gagtccaagc gagctcttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
       61 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
      121 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
      181 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
      241 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
      301 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
      361 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
      421 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
      481 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
      541 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
      601 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
      661 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
      721 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
      781 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
      841 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
      901 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
      961 ataaacaaat aggggttccg cgcgctagcg gtacccgatc ccgcgaaatg ggtaatcgct
     1021 tatcctaata cgactcacta taggccccga attcaaccgg tttgtttaac tttaagaagg
     1081 aggagtacat atggtgagca agggcgacga ggagctgttc accggggtgg tgcccatcct
     1141 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     1201 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     1261 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
     1321 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     1381 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     1441 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     1501 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     1561 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     1621 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     1681 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     1741 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     1801 gctgtacaag tccggactca gatctcgagc ggcgaacgac gaaaactact ctgaaaacta
     1861 cgcggacgcg tcttagaagc ttctagcata accccttggg gcctctaaac gggtcttgag
     1921 gggttttttg cccgggggat cccctagggc gttcggctgc ggcgagcggt atcagctcac
     1981 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     2041 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     2101 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     2161 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2221 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2281 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2341 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2401 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     2461 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     2521 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     2581 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     2641 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     2701 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgact
     2761 agtgcttgga ttctcaccaa taaaaaacgc ccggcggcaa ccgagcgttc tgaacaaatc
     2821 cagatggagt tctgaggtca ttactggatc tatcaacag
//
