LOCUS       sequence-474908-        6143 bp    DNA     circular SYN 06-JUN-2025
DEFINITION  Express SmBiT-NRF2 Fusion Protein in Mammalian Cells under a CMV 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6143)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 6143)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 6, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: SmBiT-NRF2 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6143
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    24..105
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        338..641
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        642..845
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     796..816
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          981..1113
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1096..1115
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1157..1176
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1157..1175
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3206..3327
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3217..3236)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3442..3475)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3485..3532
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3564..4358
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3618..3637)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4228..4247
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4516..5104)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4596..4615)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5230..5278
                     /note="synthetic polyadenylation signal"
     CDS             complement(5302..5700)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5697..5706
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5731..6088)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5745..5880
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5897..6088
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6045..6065
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 aagacgcgtt aattaagacg tcggcagtga aaaaaatgct ttatttgtga aatttgtgat
       61 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
      121 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
      181 ctctacaaat gtggtatggc tgattatgat ccgcggccgc aatacgtcga cgttatcagc
      241 tgacttcgta cgagagccta ggattatggc gcgccactag ttattaatag taatcaatta
      301 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      361 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      421 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      481 ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca
      541 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta
      601 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      661 acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      721 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata
      781 accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      841 gagctggttt agtgaaccgt cagatcacta gaagctttat tgcggtagtt tatcacagtt
      901 aaattgctaa cgcagtcagt gcttctgaca caacagtctc gaacttaagc tgcagaagtt
      961 ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa ggagaccaat
     1021 agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca cctattggtc
     1081 ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccagttc aattacagct
     1141 cttaaggcta gagtattaat acgactcact atagggctag cgctcaccat ggtgaccggc
     1201 taccggctgt tcgaggagat tctcgggagc tccggtggtg gcgggagcgg aggtggaggc
     1261 tcgagcggtg cgatcgccat gatggacttg gagctgccgc cgccgggact cccgtcccag
     1321 caggacatgg atttgattga catactttgg aggcaagata tagatcttgg agtaagtcga
     1381 gaagtatttg acttcagtca gcgacggaaa gagtatgagc tggaaaaaca gaaaaaactt
     1441 gaaaaggaaa gacaagaaca actccaaaag gagcaagaga aagccttttt cgctcagtta
     1501 caactagatg aagagacagg tgaatttctc ccaattcagc cagcccagca catccagtca
     1561 gaaaccagtg gatctgccaa ctactcccag gttgcccaca ttcccaaatc agatgctttg
     1621 tactttgatg actgcatgca gcttttggcg cagacattcc cgtttgtaga tgacaatgag
     1681 gtttcttcgg ctacgtttca gtcacttgtt cctgatattc ccggtcacat cgagagccca
     1741 gtcttcattg ctactaatca ggctcagtca cctgaaactt ctgttgctca ggtagcccct
     1801 gttgatttag acggtatgca acaggacatt gagcaagttt gggaggagct attatccatt
     1861 cctgagttac agtgtcttaa tattgaaaat gacaagctgg ttgagactac catggttcca
     1921 agtccagaag ccaaactgac agaagttgac aattatcatt tttactcatc tataccctca
     1981 atggaaaaag aagtaggtaa ctgtagtcca cattttctta atgcttttga ggattccttc
     2041 agcagcatcc tctccacaga agaccccaac cagttgacag tgaactcatt aaattcagat
     2101 gccacagtca acacagattt tggtgatgaa ttttattctg ctttcatagc tgagcccagt
     2161 atcagcaaca gcatgccctc acctgctact ttaagccatt cactctctga acttctaaat
     2221 gggcccattg atgtttctga tctatcactt tgcaaagctt tcaaccaaaa ccaccctgaa
     2281 agcacagcag aattcaatga ttctgactcc ggcatttcac taaacacaag tcccagtgtg
     2341 gcatcaccag aacactcagt ggaatcttcc agctatggag acacactact tggcctcagt
     2401 gattctgaag tggaagagct agatagtgcc cctggaagtg tcaaacagaa tggtcctaaa
     2461 acaccagtac attcttctgg ggatatggta caacccttgt caccatctca ggggcagagc
     2521 actcacgtgc atgatgccca atgtgagaac acaccagaga aagaattgcc tgtaagtcct
     2581 ggtcatcgga aaaccccatt cacaaaagac aaacattcaa gccgcttgga ggctcatctc
     2641 acaagagatg aacttagggc aaaagctctc catatcccat tccctgtaga aaaaatcatt
     2701 aacctccctg ttgttgactt caacgaaatg atgtccaaag agcagttcaa tgaagctcaa
     2761 cttgcattaa ttcgggatat acgtaggagg ggtaagaata aagtggctgc tcagaattgc
     2821 agaaaaagaa aactggaaaa tatagtagaa ctagagcaag atttagatca tttgaaagat
     2881 gaaaaagaaa aattgctcaa agaaaaagga gaaaatgaca aaagccttca cctactgaaa
     2941 aaacaactca gcaccttata tctcgaagtt ttcagcatgc tacgtgatga agatggaaaa
     3001 ccttattctc ctagtgaata ctccctgcag caaacaagag atggcaatgt tttccttgtt
     3061 cccaaaagta agaagccaga tgttaagaaa aacgtttaaa cgaattcggg ctcggtaccc
     3121 ggggatcctc tagagtcgac ctgcaggcat gcaagctgat ttggctgctg ccaccgctga
     3181 gcaaggccgc ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact
     3241 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     3301 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     3361 gttcaggggg agatgtggga ggttttttta agcaagtaaa acctctacaa atgtggtaaa
     3421 atcttaatta agaattctaa tataacttcg tatagcatac attatacgaa gttatggatc
     3481 cactgttgac aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag
     3541 gaactaaacc caggaggcag atcatgattg aacaagatgg attgcacgca ggttctccgg
     3601 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     3661 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     3721 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     3781 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     3841 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     3901 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     3961 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     4021 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     4081 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     4141 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     4201 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     4261 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     4321 gcatcgcctt ctatcgcctt cttgacgagt tcttctgatt cgaaatgacc gaccaagcga
     4381 cgcccaaccg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4441 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4501 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4561 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4621 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4681 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4741 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4801 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4861 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4921 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     4981 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5041 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatt
     5101 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5161 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttatagtc
     5221 cggaatcgtc acacaaaaaa ccaacacaca gatgtaatga aaataaagat attttattgc
     5281 ggccatcgtg atggctagag tttagccctc ccacacataa ccagagggca gcaattcacg
     5341 aatcccaact gccgtcggct gtccatcact gtccttcact atggctttga tcccaggatg
     5401 cagatcgaga agcacctgtc ggcaccgtcc gcaggggctc aagatgcccc tgttctcatt
     5461 tccgatcgcg acgatacaag tcaggttgcc agctgccgca gcagcagcag tgcccagcac
     5521 cacgagttct gcacaaggtc ccccagtaaa atgatataca ttgacaccag tgaagatgcg
     5581 gccgtcgcta gagagagctg cgctggcgac gctgtagtct tcagagatgg ggatgctgtt
     5641 gattgtagcc gttgctcttt caatgagggt ggattcttct tgagacaaag gcttggccat
     5701 ggtggcgcta gtgtcagaag aatcgagctt tttgcaaaag cctaggcctc caaaaaagcc
     5761 tcctcactac ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa
     5821 aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat
     5881 gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcatgc tttgcatact
     5941 tctgcctgct ggggagcctg gggactttcc acacctggtt gctgactaat tgagatgcat
     6001 gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac
     6061 attccacagc tggttctttc cgcctcagaa ggtacctaac caagttcctc tttcagaggt
     6121 tatttcaggc catggtgctg cgc
//
