LOCUS       sequence-473332-        6631 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express LgBiT-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 6631)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6631)
  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            "Unpublished"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: LgBiT-NRF2 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6631
                     /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(2685..2703)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2686..2703)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        2823..3126
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3281..3301
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        3334..3381
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3413..4207
                     /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(3467..3486)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4077..4096
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4365..4953)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4445..4464)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5079..5127
                     /note="synthetic polyadenylation signal"
     CDS             complement(5151..6188)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             5164..5172
                     /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(6219..6576)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6233..6368
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6385..6576
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6533..6553
                     /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 ctagaggatc cccgggtacc gagcccgaat tcgtttaaac
      301 gtttttctta acatctggct tcttactttt gggaacaagg aaaacattgc catctcttgt
      361 ttgctgcagg gagtattcac taggagaata aggttttcca tcttcatcac gtagcatgct
      421 gaaaacttcg agatataagg tgctgagttg ttttttcagt aggtgaaggc ttttgtcatt
      481 ttctcctttt tctttgagca atttttcttt ttcatctttc aaatgatcta aatcttgctc
      541 tagttctact atattttcca gttttctttt tctgcaattc tgagcagcca ctttattctt
      601 acccctccta cgtatatccc gaattaatgc aagttgagct tcattgaact gctctttgga
      661 catcatttcg ttgaagtcaa caacagggag gttaatgatt ttttctacag ggaatgggat
      721 atggagagct tttgccctaa gttcatctct tgtgagatga gcctccaagc ggcttgaatg
      781 tttgtctttt gtgaatgggg ttttccgatg accaggactt acaggcaatt ctttctctgg
      841 tgtgttctca cattgggcat catgcacgtg agtgctctgc ccctgagatg gtgacaaggg
      901 ttgtaccata tccccagaag aatgtactgg tgttttagga ccattctgtt tgacacttcc
      961 aggggcacta tctagctctt ccacttcaga atcactgagg ccaagtagtg tgtctccata
     1021 gctggaagat tccactgagt gttctggtga tgccacactg ggacttgtgt ttagtgaaat
     1081 gccggagtca gaatcattga attctgctgt gctttcaggg tggttttggt tgaaagcttt
     1141 gcaaagtgat agatcagaaa catcaatggg cccatttaga agttcagaga gtgaatggct
     1201 taaagtagca ggtgagggca tgctgttgct gatactgggc tcagctatga aagcagaata
     1261 aaattcatca ccaaaatctg tgttgactgt ggcatctgaa tttaatgagt tcactgtcaa
     1321 ctggttgggg tcttctgtgg agaggatgct gctgaaggaa tcctcaaaag cattaagaaa
     1381 atgtggacta cagttaccta cttctttttc cattgagggt atagatgagt aaaaatgata
     1441 attgtcaact tctgtcagtt tggcttctgg acttggaacc atggtagtct caaccagctt
     1501 gtcattttca atattaagac actgtaactc aggaatggat aatagctcct cccaaacttg
     1561 ctcaatgtcc tgttgcatac cgtctaaatc aacaggggct acctgagcaa cagaagtttc
     1621 aggtgactga gcctgattag tagcaatgaa gactgggctc tcgatgtgac cgggaatatc
     1681 aggaacaagt gactgaaacg tagccgaaga aacctcattg tcatctacaa acgggaatgt
     1741 ctgcgccaaa agctgcatgc agtcatcaaa gtacaaagca tctgatttgg gaatgtgggc
     1801 aacctgggag tagttggcag atccactggt ttctgactgg atgtgctggg ctggctgaat
     1861 tgggagaaat tcacctgtct cttcatctag ttgtaactga gcgaaaaagg ctttctcttg
     1921 ctccttttgg agttgttctt gtctttcctt ttcaagtttt ttctgttttt ccagctcata
     1981 ctctttccgt cgctgactga agtcaaatac ttctcgactt actccaagat ctatatcttg
     2041 cctccaaagt atgtcaatca aatccatgtc ctgctgggac gggagtcccg gcggcggcag
     2101 ctccaagtcc atcatggcga tcgcaccgct cgagcctcca cctccgctcc cgccaccacc
     2161 ggagctccca ctgttgatgg ttactcggaa cagcatggag ccgtcggggg tgatcaggcg
     2221 ctcgtcgata attttgttgc cgttccacag ggtccctgtt acagtgatct ttttgccgtc
     2281 gaacacggcg atgccttcat acggccgtcc gaaatagttc agcatgttcg gcgtaacccc
     2341 gtcgattacc agtgtgccat agggcaggat caccttaaag tgatgatcat ccacagggta
     2401 caccacctta aacacctctt cgatctgggc catttggtcg gcgctcagac cttcatacgg
     2461 gatgatgaca tggatgtcga tcttcagggc attttcaccg ctccggacaa tcctttggat
     2521 cggagttacg gacacggcga gattctgcag caaactggac acacctccct gttcaaggac
     2581 ttggtccagg ttgtaggcgg ctgtctgttc ccagtcccca acgaaatctt cgagtgtgaa
     2641 gaccatggtg agcgctagct gcaaaaagaa caagtagctt gtattctata gtgtcaccta
     2701 aatgggcgat ctgacggttc actaaaccag ctctgcttat ataggcctcc ctggcgcgcc
     2761 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
     2821 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
     2881 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
     2941 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
     3001 ccaagtccgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
     3061 tacatgacct tacgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
     3121 accatggtga tgcggttttg gcagtacacc aatgggcgtg gatagcggtt tgactcacgg
     3181 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
     3241 cgggactttc caaaatgtcg taataacccc gccccgttga cgcaaatggg cggtaggcgt
     3301 gtacggtggg aggtcaaatc gaattctaat actgttgaca attaatcatc ggcatagtat
     3361 atcggcatag tataatacga caaggtgagg aactaaaccc aggaggcaga tcatgattga
     3421 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     3481 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     3541 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     3601 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     3661 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     3721 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     3781 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     3841 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     3901 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     3961 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     4021 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     4081 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     4141 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     4201 cttctgattc gaaatgaccg accaagcgac gcccaaccgg tatcagctca ctcaaaggcg
     4261 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     4321 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     4381 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     4441 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     4501 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     4561 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     4621 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     4681 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     4741 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     4801 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     4861 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     4921 cagcagatta cgcgcagaaa aaaaggattt caagaagatc ctttgatctt ttctacgggg
     4981 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     5041 aggatcttca cctagatcct tttatagtcc ggaatcgtca cacaaaaaac caacacacag
     5101 atgtaatgaa aataaagata ttttattgcg gccatcgtga tggctagagt ctaaactcga
     5161 cctacctcct tagcgcgcgg tcgtgtactg ggccggcggt tgccgctgtc ggccagcacc
     5221 tcgacgcagc cgtcggtcca tacggctgcg ctccggcgag cgatttgtgt gcgaccgacg
     5281 gtgccggccc cgctgcggac gatggcatcg cagcggcctt gagcccaggc agcatcgtcg
     5341 aagttgccgt ccacgaggct ctggtagagc tgatccaggc cgatgcgcag catgtaggct
     5401 cgcagacgag ggctgccggc cagctcggga tgccggcgct cgaagtagcg agtctgctgc
     5461 tccatgcaag ccagccaggg ccgccagaag aagatgttgg ccacctcgta ctgactgtcc
     5521 ccgaacatag cttcggacca gtcgattacg gcggtgatgc ggccgttgtc ggtcaggacg
     5581 ttgttgctgc cgaagtcggc atggacgagg tggcgcacct cgggacagtc ttcggcccac
     5641 agcatcagtt cgtccagggc ttgagctacg ctggcggaca cggtgtcgtc catcacggtc
     5701 tgccagtggt agacatgggg atcagcaatg gcgcaaatga aatcccgcca ggttgtgtac
     5761 tggccgatgc cttggggccc gaatgggccg aagccgctgg tttgactgag gtcggcggcg
     5821 gcaatagcat ccatagcttc ggcgacaggc tgtaacacag caggcagctc tgtttcgggg
     5881 aggtcttgga gagtaacgcc ttgggcgcgt ctactgatgc agtatgtcag gctctcgctg
     5941 aactcgccga tgtccaacac ttcggggatg ggtagtgcag cgctggcgaa gtggcggtac
     6001 acgtagcggt ctttgtagaa gccatcagcg cagctattca cccgcagtac atagccgcgt
     6061 ccgccgacat cgaagctgaa ggctcggctc tcttcgccct ccgacaactg catcaggtcg
     6121 ctcacactgt cgaacttctc gatgagaaat ttttcaacgc tggtagcggt gagttcgggc
     6181 ttcttcatgg tggcgctagt gtcagaagaa tcgagctttt tgcaaaagcc taggcctcca
     6241 aaaaagcctc ctcactactt ctggaatagc tcagaggccg aggcggcctc ggcctctgca
     6301 taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg cggagttagg
     6361 ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag atgcatgctt
     6421 tgcatacttc tgcctgctgg ggagcctggg gactttccac acctggttgc tgactaattg
     6481 agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc acaccctaac
     6541 tgacacacat tccacagctg gttctttccg cctcagaagg tacctaacca agttcctctt
     6601 tcagaggtta tttcaggcca tggtgctgcg c
//
