LOCUS       sequence-479833-        7023 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express bRaf-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 7023)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7023)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 7, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: NanoBiT Assay Vector, bRaf
FEATURES             Location/Qualifiers
     source          1..7023
                     /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"
     regulatory      1063..1072
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3966..3984)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    4021..4142
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4032..4051)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4086..4105
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4387..4744
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4387..4578
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4410..4430)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4595..4730
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4759..4806
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4838..5632
                     /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(4892..4911)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5502..5521
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5696..5744
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(5983..6571)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6063..6082)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6687..6970
                     /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 cgccatggcg gcgctgagcg
     1081 gtggcggtgg tggcggcgcg gagccgggcc aggctctgtt caacggggac atggagcccg
     1141 aggccggcgc cggcgccggc gccgcggcct cttcggctgc ggaccctgcc attccggagg
     1201 aggtgtggaa tatcaaacaa atgattaagt tgacacagga acatatagag gccctattgg
     1261 acaaatttgg tggggagcat aatccaccat caatatatct ggaggcctat gaagaataca
     1321 ccagcaagct agatgcactc caacaaagag aacaacagtt attggaatct ctggggaacg
     1381 gaactgattt ttctgtttct agctctgcat caatggatac cgttacatct tcttcctctt
     1441 ctagcctttc agtgctacct tcatctcttt cagtttttca aaatcccaca gatgtggcac
     1501 ggagcaaccc caagtcacca caaaaaccta tcgttagagt cttcctgccc aacaaacaga
     1561 ggacagtggt acctgcaagg tgtggagtta cagtccgaga cagtctaaag aaagcactga
     1621 tgatgagagg tctaatccca gagtgctgtg ctgtttacag aattcaggat ggagagaaga
     1681 aaccaattgg ttgggacact gatatttcct ggcttactgg agaagaattg catgtggaag
     1741 tgttggagaa tgttccactt acaacacaca actttgtacg aaaaacgttt ttcaccttag
     1801 cattttgtga cttttgtcga aagctgcttt tccagggttt ccgctgtcaa acatgtggtt
     1861 ataaatttca ccagcgttgt agtacagaag ttccactgat gtgtgttaat tatgaccaac
     1921 ttgatttgct gtttgtctcc aagttctttg aacaccaccc aataccacag gaagaggcgt
     1981 ccttagcaga gactgcccta acatctggat catccccttc cgcacccgcc tcggactcta
     2041 ttgggcccca aattctcacc agtccgtctc cttcaaaatc cattccaatt ccacagccct
     2101 tccgaccagc agatgaagat catcgaaatc aatttgggca acgagaccga tcctcatcag
     2161 ctcccaatgt gcatataaac acaatagaac ctgtcaatat tgatgacttg attagagacc
     2221 aaggatttcg tggtgatgga ggatcaacca caggtttgtc tgctaccccc cctgcctcat
     2281 tacctggctc actaactaac gtgaaagcct tacagaaatc tccaggacct cagcgagaaa
     2341 ggaagtcatc ttcatcctca gaagacagga atcgaatgaa aacacttggt agacgggact
     2401 cgagtgatga ttgggagatt cctgatgggc agattacagt gggacaaaga attggatctg
     2461 gatcatttgg aacagtctac aagggaaagt ggcatggtga tgtggcagtg aaaatgttga
     2521 atgtgacagc acctacacct cagcagttac aagccttcaa aaatgaagta ggagtactca
     2581 ggaaaacacg acatgtgaat atcctactct tcatgggcta ttccacaaag ccacaactgg
     2641 ctattgttac ccagtggtgt gagggctcca gcttgtatca ccatctccat atcattgaga
     2701 ccaaatttga gatgatcaaa cttatagata ttgcacgaca gactgcacag ggcatggatt
     2761 acttacacgc caagtcaatc atccacagag acctcaagag taataatata tttcttcatg
     2821 aagacctcac agtaaaaata ggtgattttg gtctagctac agtgaaatct cgatggagtg
     2881 ggtcccatca gtttgaacag ttgtctggat ccattttgtg gatggcacca gaagtcatca
     2941 gaatgcaaga taaaaatcca tacagctttc agtcagatgt atatgcattt ggaattgttc
     3001 tgtatgaatt gatgactgga cagttacctt attcaaacat caacaacagg gaccagataa
     3061 tttttatggt gggacgagga tacctgtctc cagatctcag taaggtacgg agtaactgtc
     3121 caaaagccat gaagagatta atggcagagt gcctcaaaaa gaaaagagat gagagaccac
     3181 tctttcccca aattctcgcc tctattgagc tgctggcccg ctcattgcca aaaattcacc
     3241 gcagtgcatc agaaccctcc ttgaatcggg ctggtttcca aacagaggat tttagtctat
     3301 atgcttgtgc ttctccaaaa acacccatcc aggcaggggg atatggtgcg tttcctgtcc
     3361 acgtttctct cggaggaggt ggctcaggtg gagggggctc cgtcttcaca ctcgaagatt
     3421 tcgttgggga ctgggaacag acagccgcct acaacctgga ccaagtcctt gaacagggag
     3481 gtgtgtccag tttgctgcag aatctcgccg tgtccgtaac tccgatccaa aggattgtcc
     3541 ggagcggtga aaatgccctg aagatcgaca tccatgtcat catcccgtat gaaggtctga
     3601 gcgccgacca aatggcccag atcgaagagg tgtttaaggt ggtgtaccct gtggatgatc
     3661 atcactttaa ggtgatcctg ccctatggca cactggtaat cgacggggtt acgccgaaca
     3721 tgctgaacta tttcggacgg ccgtatgaag gcatcgccgt gttcgacggc aaaaagatca
     3781 ctgtaacagg gaccctgtgg aacggcaaca aaattatcga cgagcgcctg atcacccccg
     3841 acggctccat gctgttccga gtaaccatca acagctaagg ccgcgactct agagtcgacc
     3901 tgcaggcatg caagctgatc cggctgctaa caaagcccga aaggaagctg agttggctgc
     3961 tgccaccgct gagcaataac tagcataacc ccttggggcg gccgcttcga gcagacatga
     4021 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     4081 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     4141 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt
     4201 ttttaagcaa gtaaaacctc tacaaatgtg gtaaaatcga attttaacaa aatattaacg
     4261 cttacaattt cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     4321 tacgcggatc tgcgcagcac catggcctga aataacctct gaaagaggaa cttggttagg
     4381 taccttctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     4441 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     4501 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4561 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     4621 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     4681 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     4741 caaaaagctt aattaactgt tgacaattaa tcatcggcat agtatatcgg catagtataa
     4801 tacgacaagg tgaggaacta aacccaggag gcagatcatg attgaacaag atggattgca
     4861 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     4921 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     4981 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     5041 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     5101 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     5161 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     5221 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     5281 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     5341 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     5401 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     5461 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     5521 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     5581 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     5641 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgatg gccgcaataa
     5701 aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgaatcga tagcgataag
     5761 gatcctcttt gcgcttgcgt tttcccttgt ccagatagcc cagtagctga cattcatccg
     5821 gggtcagcac cgtttctgcg gactggcttt ctacccggta tcagctcact caaaggcggt
     5881 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     5941 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     6001 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     6061 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     6121 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     6181 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     6241 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     6301 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     6361 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     6421 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     6481 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     6541 gcagattacg cgcagaaaaa aaggatttca agaagatcct ttgatctttt ctacggggtc
     6601 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     6661 gatcttcacc tagatccttt tatagtccgg aaatacagga acgcacgctg gatggccctt
     6721 cgctgggatg gtgaaaccat gaaaaatggc agcttcagtg gattaagtgg gggtaatgtg
     6781 gcctgtaccc tctggttgca taggtattca tacggttaaa atttatcagg cgcgattgcg
     6841 gcagtttttc gggtggtttg ttgccatttt tacctgtctg ctgccgtgat cgcgctgaac
     6901 gcgttttagc ggtgcgtaca attaagggat tatggtaaat ccacttactg tctgccctcg
     6961 tagccatcga gataaaccgc agtactccgg ccacgatgcg tccggcgtag aggatcgaga
     7021 tct
//
