LOCUS       sequence-485987-        6176 bp    DNA     circular SYN 16-SEP-2025
DEFINITION  Express HiBiT-KEAP1 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 6176)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6176)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 16, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: HiBiT-KEAP1(CMV) Vector
FEATURES             Location/Qualifiers
     source          1..6176
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     94..114
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          279..411
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     394..413
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     455..474
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        455..473
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2541..2559)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2596..2717
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2607..2626)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2661..2680
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2962..3319
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2962..3153
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2985..3005)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3170..3305
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     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"
     polyA_signal    4271..4319
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4558..5146)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4638..4657)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5262..5545
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     enhancer        5736..6115
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(6116..6176,1..143)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
       61 ttccaaaatg tcgtaataac cccgccccgt tgacgcaaat gggcggtagg cgtgtacggt
      121 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatcactaga agctttattg
      181 cggtagttta tcacagttaa attgctaacg cagtcagtgc ttctgacaca acagtctcga
      241 acttaagctg cagaagttgg tcgtgaggca ctgggcaggt aagtatcaag gttacaagac
      301 aggtttaagg agaccaatag aaactgggct tgtcgagaca gagaagactc ttgcgtttct
      361 gataggcacc tattggtctt actgacatcc actttgcctt tctctccaca ggtgtccact
      421 cccagttcaa ttacagctct taaggctaga gtattaatac gactcactat agggctagcg
      481 ctcaccatgg tgagcggctg gcggctgttc aagaagatta gcgggagctc cggtggctcg
      541 agcggtgcga tcgccatgca gccagatccc aggcctagcg gggctggggc ctgctgccga
      601 ttcctgcccc tgcagtcaca gtgccctgag ggggcagggg acgcggtgat gtacgcctcc
      661 actgagtgca aggcggaggt gacgccctcc cagcatggca accgcacctt cagctacacc
      721 ctggaggatc ataccaagca ggcctttggc atcatgaacg agctgcggct cagccagcag
      781 ctgtgtgacg tcacactgca ggtcaagtac caggatgcac cggccgccca gttcatggcc
      841 cacaaggtgg tgctggcctc atccagccct gtcttcaagg ccatgttcac caacgggctg
      901 cgggagcagg gcatggaggt ggtgtccatt gagggtatcc accccaaggt catggagcgc
      961 ctcattgaat tcgcctacac ggcctccatc tccatgggcg agaagtgtgt cctccacgtc
     1021 atgaacggcg ctgtcatgta ccagatcgac agcgttgtcc gtgcctgcag tgacttcctg
     1081 gtgcagcagc tggaccccag caatgccatc ggcatcgcca acttcgctga gcagattggc
     1141 tgtgtggagt tgcaccagcg tgcccgggag tacatctaca tgcattttgg ggaggtggcc
     1201 aagcaagagg agttcttcaa cctgtcccac tgccaactgg tgaccctcat cagccgggac
     1261 gacctgaacg tgcgctgcga gtccgaggtc ttccacgcct gcatcaactg ggtcaagtac
     1321 gactgcgaac agcgacggtt ctacgtccag gcgctgctgc gggccgtgcg ctgccactcg
     1381 ttgacgccga acttcctgca gatgcagctg cagaagtgcg agatcctgca gtccgactcc
     1441 cgctgcaagg actacctggt caagatcttc gaggagctca ccctgcacaa gcccacgcag
     1501 gtgatgccct gccgggcgcc caaggtgggc cgcctgatct acaccgcggg cggctacttc
     1561 cgacagtcgc tcagctacct ggaggcttac aaccccagtg acggcacctg gctccggttg
     1621 gcggacctgc aggtgccgcg gagcggcctg gccggctgcg tggtgggcgg gctgttgtac
     1681 gccgtgggcg gcaggaacaa ctcgcccgac ggcaacaccg actccagcgc cctggactgt
     1741 tacaacccca tgaccaatca gtggtcgccc tgcgccccca tgagcgtgcc ccgtaaccgc
     1801 atcggggtgg gggtcatcga tggccacatc tatgccgtcg gcggctccca cggctgcatc
     1861 caccacaaca gtgtggagag gtatgagcca gagcgggatg agtggcactt ggtggcccca
     1921 atgctgacac gaaggatcgg ggtgggcgtg gctgtcctca atcgtctgct ttatgccgtg
     1981 gggggctttg acgggacaaa ccgccttaat tcagctgagt gttactaccc agagaggaac
     2041 gagtggcgaa tgatcacagc aatgaacacc atccgaagcg gggcaggcgt ctgcgtcctg
     2101 cacaactgta tctatgctgc tgggggctat gatggtcagg accagctgaa cagcgtggag
     2161 cgctacgatg tggaaacaga gacgtggact ttcgtagccc ccatgaagca ccggcgaagt
     2221 gccctgggga tcactgtcca ccaggggaga atctacgtcc ttggaggcta tgatggtcac
     2281 acgttcctgg acagtgtgga gtgttacgac ccagatacag acacctggag cgaggtgacc
     2341 cgaatgacat cgggccggag tggggtgggc gtggctgtca ccatggagcc ctgccggaag
     2401 cagattgacc agcagaactg tacctgtgtt taaacgaatt cgggctcggt acccggggat
     2461 cctctagagt cgacctgcag gcatgcaagc tgatccggct gctaacaaag cccgaaagga
     2521 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcggccgc
     2581 ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     2641 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     2701 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     2761 agatgtggga ggttttttta agcaagtaaa acctctacaa atgtggtaaa atcgaatttt
     2821 aacaaaatat taacgcttac aatttcctga tgcggtattt tctccttacg catctgtgcg
     2881 gtatttcaca ccgcatacgc ggatctgcgc agcaccatgg cctgaaataa cctctgaaag
     2941 aggaacttgg ttaggtacct tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     3001 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3061 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3121 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     3181 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     3241 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     3301 aggcctaggc ttttgcaaaa agcttaatta 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 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     4261 cgatggccgc aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga
     4321 atcgatagcg ataaggatcc tctttgcgct tgcgttttcc cttgtccaga tagcccagta
     4381 gctgacattc atccggggtc agcaccgttt ctgcggactg gctttctacc cggtatcagc
     4441 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4501 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4561 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4621 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4681 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4741 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4801 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4861 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4921 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4981 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5041 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5101 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tttcaagaag atcctttgat
     5161 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5221 gagattatca aaaaggatct tcacctagat ccttttatag tccggaaata caggaacgca
     5281 cgctggatgg cccttcgctg ggatggtgaa accatgaaaa atggcagctt cagtggatta
     5341 agtgggggta atgtggcctg taccctctgg ttgcataggt attcatacgg ttaaaattta
     5401 tcaggcgcga ttgcggcagt ttttcgggtg gtttgttgcc atttttacct gtctgctgcc
     5461 gtgatcgcgc tgaacgcgtt ttagcggtgc gtacaattaa gggattatgg taaatccact
     5521 tactgtctgc cctcgtagcc atcgagataa accgcagtac tccggccacg atgcgtccgg
     5581 cgtagaggat cgagatcttc aatattggcc attagccata ttattcattg gttatatagc
     5641 ataaatcaat attggctatt ggccattgca tacgttgtat ctatatcata atatgtacat
     5701 ttatattggc tcatgtccaa tatgaccgcc atgttggcat tgattattga ctagttatta
     5761 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     5821 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     5881 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     5941 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtccgcc
     6001 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     6061 acgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
     6121 gcggttttgg cagtacacca atgggcgtgg atagcggttt gactcacggg gatttc
//
