LOCUS       sequence-341149-        2445 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  QUAS directly upstream of a T7 promoter. Expresses CcdB 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 2445)
  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 2445)
  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-CcdB_ColE1
FEATURES             Location/Qualifiers
     source          1..2445
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(6..797,798..866))
                     /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     629..648
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(867..971)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1015..1033
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1051..1071
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1080..1385
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MIQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSD
                     KVSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     1267..1286
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     terminator      1458..1505
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      complement(1669..2257)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1749..1768)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2345..2439)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
ORIGIN
        1 agctcttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
       61 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
      121 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
      181 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
      241 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
      301 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
      361 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
      421 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
      481 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
      541 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
      601 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
      661 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
      721 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
      781 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
      841 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
      901 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
      961 ggggttccgc gcgctagcga attccgatcc cgcgaaatgg gtaatcgctt atcctaatac
     1021 gactcactat aggcccctct agaaataatt ttgtttaact ttaagaagga ggagtacata
     1081 tgattcagtt taaggtttac acctataaaa gagagagccg ttatcgtctg tttgtggatg
     1141 tacagagtga tattattgac acgcccgggc gacggatggt gatccccctg gccagtgcac
     1201 gtctgctgtc agataaagtc tcccgtgaac tttacccggt ggtgcatatc ggggatgaaa
     1261 gctggcgcat gatgaccacc gatatggcca gtgtgccggt ctccgttatc ggggaagaag
     1321 tggctgatct cagccaccgc gaaaatgaca tcaaaaacgc cattaacctg atgttctggg
     1381 gaatatccgg actcagatct cgagcggcga acgacgaaaa ctactctgaa aactacgcgg
     1441 acgcgtctta aaagcttcta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
     1501 ttttgcccgg gggatcccct agggcgttcg gctgcggcga gcggtatcag ctcactcaaa
     1561 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     1621 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     1681 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     1741 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     1801 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     1861 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     1921 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     1981 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     2041 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     2101 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     2161 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     2221 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     2281 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgactagtgc
     2341 ttggattctc accaataaaa aacgcccggc ggcaaccgag cgttctgaac aaatccagat
     2401 ggagttctga ggtcattact ggatctatca acaggagtcc aagcg
//
