LOCUS       sequence-474907-        6605 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express NRF2-LgBiT 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 6605)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6605)
  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: 
            "Unpublished"
COMMENT     Sequence Label: NRF2-LgBiT Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6605
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    17..98
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     42..61
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(2659..2677)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2660..2677)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        2797..3100
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3255..3275
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        3308..3355
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3387..4181
                     /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(3441..3460)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4051..4070
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4339..4927)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4419..4438)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5053..5101
                     /note="synthetic polyadenylation signal"
     CDS             complement(5125..6162)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             5138..5146
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     promoter        complement(6193..6550)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6207..6342
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6359..6550
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6507..6527
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 aagacgcgtg acgtcggcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
       61 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
      121 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
      181 aatgtggtat ggctgattat gatccgcggc cgcaatacgt cgacgttatc agctgacttc
      241 gtacgagagc ctaggtgttt ctagagtcgc ggccttagct gttgatggtt actcggaaca
      301 gcatggagcc gtcgggggtg atcaggcgct cgtcgataat tttgttgccg ttccacaggg
      361 tccctgttac agtgatcttt ttgccgtcga acacggcgat gccttcatac ggccgtccga
      421 aatagttcag catgttcggc gtaaccccgt cgattaccag tgtgccatag ggcaggatca
      481 ccttaaagtg atgatcatcc acagggtaca ccaccttaaa cacctcttcg atctgggcca
      541 tttggtcggc gctcagacct tcatacggga tgatgacatg gatgtcgatc ttcagggcat
      601 tttcaccgct ccggacaatc ctttggatcg gagttacgga cacggcgaga ttctgcagca
      661 aactggacac acctccctgt tcaaggactt ggtccaggtt gtaggcggct gtctgttccc
      721 agtccccaac gaaatcttcg agtgtgaaga caccgctcga gcctccacct ccgctcccgc
      781 caccacctga actgccttga gaaacgtttt tcttaacatc tggcttctta cttttgggaa
      841 caaggaaaac attgccatct cttgtttgct gcagggagta ttcactagga gaataaggtt
      901 ttccatcttc atcacgtagc atgctgaaaa cttcgagata taaggtgctg agttgttttt
      961 tcagtaggtg aaggcttttg tcattttctc ctttttcttt gagcaatttt tctttttcat
     1021 ctttcaaatg atctaaatct tgctctagtt ctactatatt ttccagtttt ctttttctgc
     1081 aattctgagc agccacttta ttcttacccc tcctacgtat atcccgaatt aatgcaagtt
     1141 gagcttcatt gaactgctct ttggacatca tttcgttgaa gtcaacaaca gggaggttaa
     1201 tgattttttc tacagggaat gggatatgga gagcttttgc cctaagttca tctcttgtga
     1261 gatgagcctc caagcggctt gaatgtttgt cttttgtgaa tggggttttc cgatgaccag
     1321 gacttacagg caattctttc tctggtgtgt tctcacattg ggcatcatgc acgtgagtgc
     1381 tctgcccctg agatggtgac aagggttgta ccatatcccc agaagaatgt actggtgttt
     1441 taggaccatt ctgtttgaca cttccagggg cactatctag ctcttccact tcagaatcac
     1501 tgaggccaag tagtgtgtct ccatagctgg aagattccac tgagtgttct ggtgatgcca
     1561 cactgggact tgtgtttagt gaaatgccgg agtcagaatc attgaattct gctgtgcttt
     1621 cagggtggtt ttggttgaaa gctttgcaaa gtgatagatc agaaacatca atgggcccat
     1681 ttagaagttc agagagtgaa tggcttaaag tagcaggtga gggcatgctg ttgctgatac
     1741 tgggctcagc tatgaaagca gaataaaatt catcaccaaa atctgtgttg actgtggcat
     1801 ctgaatttaa tgagttcact gtcaactggt tggggtcttc tgtggagagg atgctgctga
     1861 aggaatcctc aaaagcatta agaaaatgtg gactacagtt acctacttct ttttccattg
     1921 agggtataga tgagtaaaaa tgataattgt caacttctgt cagtttggct tctggacttg
     1981 gaaccatggt agtctcaacc agcttgtcat tttcaatatt aagacactgt aactcaggaa
     2041 tggataatag ctcctcccaa acttgctcaa tgtcctgttg cataccgtct aaatcaacag
     2101 gggctacctg agcaacagaa gtttcaggtg actgagcctg attagtagca atgaagactg
     2161 ggctctcgat gtgaccggga atatcaggaa caagtgactg aaacgtagcc gaagaaacct
     2221 cattgtcatc tacaaacggg aatgtctgcg ccaaaagctg catgcagtca tcaaagtaca
     2281 aagcatctga tttgggaatg tgggcaacct gggagtagtt ggcagatcca ctggtttctg
     2341 actggatgtg ctgggctggc tgaattggga gaaattcacc tgtctcttca tctagttgta
     2401 actgagcgaa aaaggctttc tcttgctcct tttggagttg ttcttgtctt tccttttcaa
     2461 gttttttctg tttttccagc tcatactctt tccgtcgctg actgaagtca aatacttctc
     2521 gacttactcc aagatctata tcttgcctcc aaagtatgtc aatcaaatcc atgtcctgct
     2581 gggacgggag tcccggcggc ggcagctcca agtccatcat ggcgatcgct agctgcaaaa
     2641 agaacaagta gcttgtattc tatagtgtca cctaaatggg cgatctgacg gttcactaaa
     2701 ccagctctgc ttatataggc ctccctggcg cgccactagt tattaatagt aatcaattac
     2761 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     2821 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     2881 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     2941 tgcccacttg gcagtacatc aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa
     3001 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttacggg actttcctac
     3061 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
     3121 caccaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     3181 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa
     3241 ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtca aatcgaattc
     3301 taatactgtt gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt
     3361 gaggaactaa acccaggagg cagatcatga ttgaacaaga tggattgcac gcaggttctc
     3421 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     3481 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     3541 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     3601 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     3661 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     3721 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     3781 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     3841 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     3901 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     3961 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     4021 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     4081 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     4141 agcgcatcgc cttctatcgc cttcttgacg agttcttctg attcgaaatg accgaccaag
     4201 cgacgcccaa ccggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4261 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4321 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4381 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4441 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4501 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4561 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4621 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4681 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4741 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     4801 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4861 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4921 atttcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4981 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttata
     5041 gtccggaatc gtcacacaaa aaaccaacac acagatgtaa tgaaaataaa gatattttat
     5101 tgcggccatc gtgatggcta gagtctaaac tcgacctacc tccttagcgc gcggtcgtgt
     5161 actgggccgg cggttgccgc tgtcggccag cacctcgacg cagccgtcgg tccatacggc
     5221 tgcgctccgg cgagcgattt gtgtgcgacc gacggtgccg gccccgctgc ggacgatggc
     5281 atcgcagcgg ccttgagccc aggcagcatc gtcgaagttg ccgtccacga ggctctggta
     5341 gagctgatcc aggccgatgc gcagcatgta ggctcgcaga cgagggctgc cggccagctc
     5401 gggatgccgg cgctcgaagt agcgagtctg ctgctccatg caagccagcc agggccgcca
     5461 gaagaagatg ttggccacct cgtactgact gtccccgaac atagcttcgg accagtcgat
     5521 tacggcggtg atgcggccgt tgtcggtcag gacgttgttg ctgccgaagt cggcatggac
     5581 gaggtggcgc acctcgggac agtcttcggc ccacagcatc agttcgtcca gggcttgagc
     5641 tacgctggcg gacacggtgt cgtccatcac ggtctgccag tggtagacat ggggatcagc
     5701 aatggcgcaa atgaaatccc gccaggttgt gtactggccg atgccttggg gcccgaatgg
     5761 gccgaagccg ctggtttgac tgaggtcggc ggcggcaata gcatccatag cttcggcgac
     5821 aggctgtaac acagcaggca gctctgtttc ggggaggtct tggagagtaa cgccttgggc
     5881 gcgtctactg atgcagtatg tcaggctctc gctgaactcg ccgatgtcca acacttcggg
     5941 gatgggtagt gcagcgctgg cgaagtggcg gtacacgtag cggtctttgt agaagccatc
     6001 agcgcagcta ttcacccgca gtacatagcc gcgtccgccg acatcgaagc tgaaggctcg
     6061 gctctcttcg ccctccgaca actgcatcag gtcgctcaca ctgtcgaact tctcgatgag
     6121 aaatttttca acgctggtag cggtgagttc gggcttcttc atggtggcgc tagtgtcaga
     6181 agaatcgagc tttttgcaaa agcctaggcc tccaaaaaag cctcctcact acttctggaa
     6241 tagctcagag gccgaggcgg cctcggcctc tgcataaata aaaaaaatta gtcagccatg
     6301 gggcggagaa tgggcggaac tgggcggagt taggggcggg atgggcggag ttaggggcgg
     6361 gactatggtt gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc
     6421 tggggacttt ccacacctgg ttgctgacta attgagatgc atgctttgca tacttctgcc
     6481 tgctggggag cctggggact ttccacaccc taactgacac acattccaca gctggttctt
     6541 tccgcctcag aaggtaccta accaagttcc tctttcagag gttatttcag gccatggtgc
     6601 tgcgc
//
