LOCUS       sequence-475086-        6150 bp    DNA     circular SYN 23-JUL-2025
DEFINITION  Express KEAP1-HiBiT 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 6150)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6150)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 23, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: KEAP1-HiBiT(CMV) Vector
FEATURES             Location/Qualifiers
     source          1..6150
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1033..1052
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1033..1051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3093..3111)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3148..3269
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3159..3178)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3213..3232
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3514..3871
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3514..3705
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3537..3557)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3722..3857
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3886..3933
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3965..4759
                     /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(4019..4038)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4629..4648
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4823..4871
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(5110..5698)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5190..5209)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5814..6097
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tggtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt attaatacga ctcactatag ggctagcgat cgccatgcag ccagatccca
     1081 ggcctagcgg ggctggggcc tgctgccgat tcctgcccct gcagtcacag tgccctgagg
     1141 gggcagggga cgcggtgatg tacgcctcca ctgagtgcaa ggcggaggtg acgccctccc
     1201 agcatggcaa ccgcaccttc agctacaccc tggaggatca taccaagcag gcctttggca
     1261 tcatgaacga gctgcggctc agccagcagc tgtgtgacgt cacactgcag gtcaagtacc
     1321 aggatgcacc ggccgcccag ttcatggccc acaaggtggt gctggcctca tccagccctg
     1381 tcttcaaggc catgttcacc aacgggctgc gggagcaggg catggaggtg gtgtccattg
     1441 agggtatcca ccccaaggtc atggagcgcc tcattgaatt cgcctacacg gcctccatct
     1501 ccatgggcga gaagtgtgtc ctccacgtca tgaacggcgc tgtcatgtac cagatcgaca
     1561 gcgttgtccg tgcctgcagt gacttcctgg tgcagcagct ggaccccagc aatgccatcg
     1621 gcatcgccaa cttcgctgag cagattggct gtgtggagtt gcaccagcgt gcccgggagt
     1681 acatctacat gcattttggg gaggtggcca agcaagagga gttcttcaac ctgtcccact
     1741 gccaactggt gaccctcatc agccgggacg acctgaacgt gcgctgcgag tccgaggtct
     1801 tccacgcctg catcaactgg gtcaagtacg actgcgaaca gcgacggttc tacgtccagg
     1861 cgctgctgcg ggccgtgcgc tgccactcgt tgacgccgaa cttcctgcag atgcagctgc
     1921 agaagtgcga gatcctgcag tccgactccc gctgcaagga ctacctggtc aagatcttcg
     1981 aggagctcac cctgcacaag cccacgcagg tgatgccctg ccgggcgccc aaggtgggcc
     2041 gcctgatcta caccgcgggc ggctacttcc gacagtcgct cagctacctg gaggcttaca
     2101 accccagtaa cggcacctgg ctccggttgg cggacctgca ggtgccgcgg agcggcctgg
     2161 ccggctgcgt ggtgggcggg ctgttgtacg ccgtgggcgg caggaacaac tcgcccgacg
     2221 gcaacaccga ctccagcgcc ctggactgtt acaaccccat gaccaatcag tggtcgccct
     2281 gcgcccccat gagcgtgccc cgtaaccgca tcggggtggg ggtcatcgat ggccacatct
     2341 atgccgtcgg cggctcccac ggctgcatcc accacaacag tgtggagagg tatgagccag
     2401 agcgggatga gtggcacttg gtggccccaa tgctgacacg aaggatcggg gtgggcgtgg
     2461 ctgtcctcaa tcgtctgctt tatgccgtgg ggggctttga cgggacaaac cgccttaatt
     2521 cagctgagtg ttactaccca gagaggaacg agtggcgaat gatcacagca atgaacacca
     2581 tccgaagcgg ggcaggcgtc tgcgtcctgc acaactgtat ctatgctgct gggggctatg
     2641 atggtcagga ccagctgaac agcgtggagc gctacgatgt ggaaacagag acgtggactt
     2701 tcgtagcccc catgaagcac cggcgaagtg ccctggggat cactgtccac caggggagaa
     2761 tctacgtcct tggaggctat gatggtcaca cgttcctgga cagtgtggag tgttacgacc
     2821 cagatacaga cacctggagc gaggtgaccc gaatgacatc gggccggagt ggggtgggcg
     2881 tggctgtcac catggagccc tgccggaagc agattgacca gcagaactgt acctgtgttt
     2941 ctcaaggcag ttcaggtggc tcgagcggtg tgagcggctg gcggctgttc aagaagatta
     3001 gctaaggccg cgactctaga gtcgacctgc aggcatgcaa gctgatccgg ctgctaacaa
     3061 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct
     3121 tggggcggcc gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa
     3181 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     3241 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     3301 aggttcaggg ggagatgtgg gaggtttttt taagcaagta aaacctctac aaatgtggta
     3361 aaatcgaatt ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta
     3421 cgcatctgtg cggtatttca caccgcatac gcggatctgc gcagcaccat ggcctgaaat
     3481 aacctctgaa agaggaactt ggttaggtac cttctgaggc ggaaagaacc agctgtggaa
     3541 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3601 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     3661 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     3721 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     3781 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     3841 aggctttttt ggaggcctag gcttttgcaa aaagcttaat taactgttga caattaatca
     3901 tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac ccaggaggca
     3961 gatcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     4021 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     4081 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     4141 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     4201 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     4261 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     4321 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     4381 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     4441 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     4501 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     4561 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     4621 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     4681 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     4741 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     4801 aacctgccat cacgatggcc gcaataaaat atctttattt tcattacatc tgtgtgttgg
     4861 ttttttgtgt gaatcgatag cgataaggat cctctttgcg cttgcgtttt cccttgtcca
     4921 gatagcccag tagctgacat tcatccgggg tcagcaccgt ttctgcggac tggctttcta
     4981 cccggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     5041 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     5101 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     5161 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5221 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     5281 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     5341 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     5401 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     5461 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     5521 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     5581 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     5641 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatttcaaga
     5701 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     5761 gattttggtc atgagattat caaaaaggat cttcacctag atccttttat agtccggaaa
     5821 tacaggaacg cacgctggat ggcccttcgc tgggatggtg aaaccatgaa aaatggcagc
     5881 ttcagtggat taagtggggg taatgtggcc tgtaccctct ggttgcatag gtattcatac
     5941 ggttaaaatt tatcaggcgc gattgcggca gtttttcggg tggtttgttg ccatttttac
     6001 ctgtctgctg ccgtgatcgc gctgaacgcg ttttagcggt gcgtacaatt aagggattat
     6061 ggtaaatcca cttactgtct gccctcgtag ccatcgagat aaaccgcagt actccggcca
     6121 cgatgcgtcc ggcgtagagg atcgagatct
//
