LOCUS       sequence-475087-        6176 bp    DNA     circular SYN 23-JUL-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  JOURNAL   Unpublished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 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            "Unpublished"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: HiBiT-KEAP1(CMV) Vector
FEATURES             Location/Qualifiers
     source          1..6176
                     /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(3119..3137)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3174..3295
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3185..3204)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3239..3258
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3540..3897
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3540..3731
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3563..3583)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3748..3883
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3912..3959
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3991..4785
                     /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(4045..4064)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4655..4674
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4849..4897
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(5136..5724)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5216..5235)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5840..6123
                     /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 ggctagcgct caccatggtg agcggctggc
     1081 ggctgttcaa gaagattagc gggagctccg gtggctcgag cggtgcgatc gccatgcagc
     1141 cagatcccag gcctagcggg gctggggcct gctgccgatt cctgcccctg cagtcacagt
     1201 gccctgaggg ggcaggggac gcggtgatgt acgcctccac tgagtgcaag gcggaggtga
     1261 cgccctccca gcatggcaac cgcaccttca gctacaccct ggaggatcat accaagcagg
     1321 cctttggcat catgaacgag ctgcggctca gccagcagct gtgtgacgtc acactgcagg
     1381 tcaagtacca ggatgcaccg gccgcccagt tcatggccca caaggtggtg ctggcctcat
     1441 ccagccctgt cttcaaggcc atgttcacca acgggctgcg ggagcagggc atggaggtgg
     1501 tgtccattga gggtatccac cccaaggtca tggagcgcct cattgaattc gcctacacgg
     1561 cctccatctc catgggcgag aagtgtgtcc tccacgtcat gaacggcgct gtcatgtacc
     1621 agatcgacag cgttgtccgt gcctgcagtg acttcctggt gcagcagctg gaccccagca
     1681 atgccatcgg catcgccaac ttcgctgagc agattggctg tgtggagttg caccagcgtg
     1741 cccgggagta catctacatg cattttgggg aggtggccaa gcaagaggag ttcttcaacc
     1801 tgtcccactg ccaactggtg accctcatca gccgggacga cctgaacgtg cgctgcgagt
     1861 ccgaggtctt ccacgcctgc atcaactggg tcaagtacga ctgcgaacag cgacggttct
     1921 acgtccaggc gctgctgcgg gccgtgcgct gccactcgtt gacgccgaac ttcctgcaga
     1981 tgcagctgca gaagtgcgag atcctgcagt ccgactcccg ctgcaaggac tacctggtca
     2041 agatcttcga ggagctcacc ctgcacaagc ccacgcaggt gatgccctgc cgggcgccca
     2101 aggtgggccg cctgatctac accgcgggcg gctacttccg acagtcgctc agctacctgg
     2161 aggcttacaa ccccagtaac ggcacctggc tccggttggc ggacctgcag gtgccgcgga
     2221 gcggcctggc cggctgcgtg gtgggcgggc tgttgtacgc cgtgggcggc aggaacaact
     2281 cgcccgacgg caacaccgac tccagcgccc tggactgtta caaccccatg accaatcagt
     2341 ggtcgccctg cgcccccatg agcgtgcccc gtaaccgcat cggggtgggg gtcatcgatg
     2401 gccacatcta tgccgtcggc ggctcccacg gctgcatcca ccacaacagt gtggagaggt
     2461 atgagccaga gcgggatgag tggcacttgg tggccccaat gctgacacga aggatcgggg
     2521 tgggcgtggc tgtcctcaat cgtctgcttt atgccgtggg gggctttgac gggacaaacc
     2581 gccttaattc agctgagtgt tactacccag agaggaacga gtggcgaatg atcacagcaa
     2641 tgaacaccat ccgaagcggg gcaggcgtct gcgtcctgca caactgtatc tatgctgctg
     2701 ggggctatga tggtcaggac cagctgaaca gcgtggagcg ctacgatgtg gaaacagaga
     2761 cgtggacttt cgtagccccc atgaagcacc ggcgaagtgc cctggggatc actgtccacc
     2821 aggggagaat ctacgtcctt ggaggctatg atggtcacac gttcctggac agtgtggagt
     2881 gttacgaccc agatacagac acctggagcg aggtgacccg aatgacatcg ggccggagtg
     2941 gggtgggcgt ggctgtcacc atggagccct gccggaagca gattgaccag cagaactgta
     3001 cctgtgttta aacgaattcg ggctcggtac ccggggatcc tctagagtcg acctgcaggc
     3061 atgcaagctg atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc
     3121 gctgagcaat aactagcata accccttggg gcggccgctt cgagcagaca tgataagata
     3181 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     3241 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     3301 caacaattgc attcatttta tgtttcaggt tcagggggag atgtgggagg tttttttaag
     3361 caagtaaaac ctctacaaat gtggtaaaat cgaattttaa caaaatatta acgcttacaa
     3421 tttcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgcgg
     3481 atctgcgcag caccatggcc tgaaataacc tctgaaagag gaacttggtt aggtaccttc
     3541 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     3601 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     3661 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3721 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     3781 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     3841 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     3901 cttaattaac tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca
     3961 aggtgaggaa ctaaacccag gaggcagatc atgattgaac aagatggatt gcacgcaggt
     4021 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     4081 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     4141 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     4201 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     4261 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     4321 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     4381 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     4441 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     4501 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     4561 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     4621 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     4681 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     4741 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     4801 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg atggccgcaa taaaatatct
     4861 ttattttcat tacatctgtg tgttggtttt ttgtgtgaat cgatagcgat aaggatcctc
     4921 tttgcgcttg cgttttccct tgtccagata gcccagtagc tgacattcat ccggggtcag
     4981 caccgtttct gcggactggc tttctacccg gtatcagctc actcaaaggc ggtaatacgg
     5041 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     5101 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5161 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5221 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5281 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     5341 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5401 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5461 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5521 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     5581 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5641 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5701 acgcgcagaa aaaaaggatt tcaagaagat cctttgatct tttctacggg gtctgacgct
     5761 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5821 acctagatcc ttttatagtc cggaaataca ggaacgcacg ctggatggcc cttcgctggg
     5881 atggtgaaac catgaaaaat ggcagcttca gtggattaag tgggggtaat gtggcctgta
     5941 ccctctggtt gcataggtat tcatacggtt aaaatttatc aggcgcgatt gcggcagttt
     6001 ttcgggtggt ttgttgccat ttttacctgt ctgctgccgt gatcgcgctg aacgcgtttt
     6061 agcggtgcgt acaattaagg gattatggta aatccactta ctgtctgccc tcgtagccat
     6121 cgagataaac cgcagtactc cggccacgat gcgtccggcg tagaggatcg agatct
//
