LOCUS       sequence-350155-        2850 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  QUAS directly downstream 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 2850)
  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 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"; journalName: "Nat 
            Commun."; date: "2021-07-5"; volume: "12(1):4109. doi"; pages: " 
            10.1038/s41467-021-24434-9"
COMMENT     Sequence Label: T7-0-QUAS-GFP_ColE1
FEATURES             Location/Qualifiers
     source          1..2850
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     157..176
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     416..437
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     RBS             491..511
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(544..563)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(545..563)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        585..689
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(690..758,759..1550)
                     /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     complement(908..927)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      1562..1656
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      1744..2332
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2233..2252
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2496..2543)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(join(2616..2850,1..476,477..479,480..482))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(2616..2850,1..476,477..479,480..482))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2658..2679)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 ccgttcttct gcttgtcggc catgatatag acgttgtggc tgttgtagtt gtactccagc
       61 ttgtgcccca ggatgttgcc gtcctccttg aagtcgatgc ccttcagctc gatgcggttc
      121 accagggtgt cgccctcgaa cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg
      181 aagaagatgg tgcgctcctg gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc
      241 tgcttcatgt ggtcggggta gcggctgaag cactgcacgc cgtaggtcag ggtggtcacg
      301 agggtgggcc agggcacggg cagcttgccg gtggtgcaga tgaacttcag ggtcagcttg
      361 ccgtaggtgg catcgccctc gccctcgccg gacacgctga acttgtggcc gtttacgtcg
      421 ccgtccagct cgaccaggat gggcaccacc ccggtgaaca gctcctcgcc cttgctcacc
      481 atatgtactc ctccttctta aagttaaaca aaattatttc tagagggggg ataagcgatt
      541 accccctata gtgagtcgta ttaatttcgc gggatcggct agcgcgcgga acccctattt
      601 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
      661 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
      721 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
      781 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
      841 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
      901 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
      961 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     1021 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     1081 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     1141 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     1201 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     1261 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     1321 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     1381 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     1441 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     1501 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa gagctcgctt
     1561 ggactcctgt tgatagatcc agtaatgacc tcagaactcc atctggattt gttcagaacg
     1621 ctcggttgcc gccgggcgtt ttttattggt gagaatccaa gcactagtca tgaccaaaat
     1681 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     1741 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     1801 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     1861 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     1921 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     1981 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     2041 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     2101 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     2161 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     2221 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     2281 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     2341 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     2401 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     2461 tcgccgcagc cgaacgccct aggggatccc ccgggcaaaa aacccctcaa gacccgttta
     2521 gaggccccaa ggggttatgc tagaagcttc taagacgcgt ccgcgtagtt ttcagagtag
     2581 ttttcgtcgt tcgccgctcg agatctgagt ccggacttgt acagctcgtc catgccgaga
     2641 gtgatcccgg cggcggtcac gaactccagc aggaccatgt gatcgcgctt ctcgttgggg
     2701 tctttgctca gggcggactg ggtgctcagg tagtggttgt cgggcagcag cacggggccg
     2761 tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct gcacgctgcc gtcctcgatg
     2821 ttgtggcgga tcttgaagtt caccttgatg
//
