LOCUS       sequence-478039-        4621 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expressing a grazoprevir-inducible transcriptional 
            activator.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4621)
  AUTHORS   Peterman EL, Ploessl DS, Love KS, Sanabria V, Daniels RF, Johnstone 
            CP, Godavarti DR, Kabaria SR, Oakes CG, Pai AA, Galloway KE
  TITLE     High-resolution profiling reveals coupled transcriptional and 
            translational regulation of transgenes.
  JOURNAL   Nucleic Acids Research, Volume 53, Issue 11, 24 June 2025, gkaf528
  PUBMED    39651241
REFERENCE   2  (bases 1 to 4621)
  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: "Nucleic 
            Acids Research, Volume 53, Issue 11, 24 June 2025, gkaf528"
COMMENT     Sequence Label: pKG2309_pGEEC524_pShip-EFS-ZF10-NS3-p65-bGH
FEATURES             Location/Qualifiers
     source          1..4621
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(111..130)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             122..931
                     /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="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPGDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(189..208)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1021..1609
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1510..1529
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      1774..1860
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1952..1979
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     1987..2010
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     promoter        2047..2258
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             2811..2876
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2883..2903
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3753..3779
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4005..4376
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALLSQISS"
     polyA_signal    4397..4621
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 cgctcgctga ctgcaggtgc ccgtgtctca aaatctctga tgttacattg cacaagataa
       61 aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt
      121 tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga
      181 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
      241 cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
      301 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc
      361 gaccatcaag cattttatcc gtactcctgg tgatgcatgg ttactcacca ctgcgatccc
      421 cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
      481 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
      541 cagcgatcgc gtatttcgtc tagctcaggc gcaatcacga atgaataacg gtttggttga
      601 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
      661 gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
      721 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
      781 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
      841 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
      901 gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacatta
      961 ttcagattgg gccccgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     1021 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     1081 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     1141 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     1201 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     1261 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     1321 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     1381 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     1441 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     1501 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     1561 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     1621 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     1681 gttatcccct gattctgtgg ataaccgtat taccgctagc atggatctcg gggacgtcta
     1741 actactaagc gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcggaa
     1801 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat
     1861 ccgccgggag cggatttgaa cgttgtgaag caacggcccg gagggtggcg ggcaggacgc
     1921 ccgccataaa ctgccaggca tcaaactaag cagaaggcca tcctgacgga tggccttttt
     1981 gcgtttctac aaactcttcc tgttagttag ttacttaagc tcgggcccac ctgcagtcta
     2041 cttactgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca
     2101 attgaaccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact
     2161 ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga
     2221 acgttctttt tcgcaacggg tttgccgcca gaacacagta atgagccacc atgtctagac
     2281 ccggagagcg cccattccag tgtcggattt gcatgcggaa cttttcgaga agacacggcc
     2341 tggacagaca tacccgtact catacaggtg aaaaaccctt tcagtgtcgg atctgtatgc
     2401 gaaatttctc cgaccacagc agcctgaaga gacatctacg tacccacacc ggcagccaga
     2461 agccatttca gtgtcggatc tgtatgcgga acttctccgt gagacacaac ctgaccagac
     2521 atctacgtac gcacaccgga gagaagccat tccaatgccg aatatgcatg cgcaacttca
     2581 gtgaccacag caacctgagc agacacctaa aaacccacac cggttcccag aagccatttc
     2641 agtgtcggat ctgtatgcgg aacttctccc agcgcagcag cctggtgaga catctacgta
     2701 cgcacaccgg agagaagcca ttccaatgcc gaatatgcat gcgcaacttc agtgagagcg
     2761 gccacctgaa gagacacctg cgtacgcacc tgaggggatc cacctgcagg gactacaaag
     2821 accatgacgg tgattataaa gatcatgaca tcgattacaa ggatgacgat gacaagatgg
     2881 cccccaagaa aaagaggaag gtgggcattc acggggtgcc gggtggactc gagggaggcg
     2941 gtggaagcgg cggtaccgag gacgtggtgt gctgccactc aatctacggc aagaagaagg
     3001 gtgatatcga cacctaccga tacataggct cttccgggac aggctgcgtg gtcatagtgg
     3061 gcaggatcgt cttgtccgga tccggcacta gtgcgcccat cacggcgtac gcccagcaga
     3121 cgagaggcct cctagggtgt ataatcacca gcctgactgg ccgggacaaa aaccaagtgg
     3181 agggtgaggt ccagatcgtg tcaactgcta cccaaacctt cctggcaacg tgcatcaatg
     3241 gggtatgctg ggcagtctac cacggggccg gaacgaggac catcgcatca cccaagggtc
     3301 ctgtcatcca gatgtatacc aatgtggacc aagaccttgt gggctggccc gctcctcaag
     3361 gttcccgctc attgacaccc tgtacctgcg gctcctcgga cctttacctg gtcacgaggc
     3421 acgccgatgt cattcccgtg cgccggcgag gtgatagcag gggtagcctg ctttcgcccc
     3481 ggcccatttc ctacttgaaa ggctcctcgg ggggtccgct gttgtgcccc gcgggacacg
     3541 ccgtgggcct attcagggcc gcggtgtgca cccgtggagt ggctaaagcg gtggacttta
     3601 tccctgtgga gaacctagag acaaccatga gatccccggt gttcacggac aactcctctc
     3661 caccagcagt caccctgacg cacccaatca ccaaaatcga tagggaggtt ctctaccagg
     3721 agttcgatga gatggaagag tgctctcagc actatcccta cgatgtgccc gattacgctg
     3781 gaggcggtgg aagcggcggt accgatgagt ttcccaccat ggtgtttcct tctgggcaga
     3841 tcagccaggc ctcggccttg gccccggccc ctccccaagt cctgccccag gctccagccc
     3901 ctgcccctgc tccagccatg gtatcagctc tggcccaggc cccagcccct gtcccagtcc
     3961 tagccccagg ccctcctcag gctgtggccc cacctgcccc caagcccacc caggctgggg
     4021 aaggaacgct gtcagaggcc ctgctgcagc tgcagtttga tgatgaagac ctgggggcct
     4081 tgcttggcaa cagcacagac ccagctgtgt tcacagacct ggcatccgtc gacaactccg
     4141 agtttcagca gctgctgaac cagggcatac ctgtggcccc ccacacaact gagcccatgc
     4201 tgatggagta ccctgaggct ataactcgcc tagtgacagg ggcccagagg ccccccgacc
     4261 cagctcctgc tccactgggg gccccggggc tccccaatgg cctcctttca ggagatgaag
     4321 acttctcctc cattgcggac atggacttct cagccctgct gagtcagatc agctcccaat
     4381 tgtaacaacg ttgcacctgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     4441 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     4501 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     4561 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     4621 g
//
