LOCUS       sequence-481969-        6090 bp    DNA     circular SYN 05-AUG-2025
DEFINITION  Express HaloTag-KRAS(G12V) 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 6090)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6090)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 5, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: NanoBRET Assay Vector, HaloTag-KRAS(G12V)
FEATURES             Location/Qualifiers
     source          1..6090
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        74..453
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    644..927
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     rep_origin      1043..1631
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1532..1551
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    1870..1918
                     /note="synthetic polyadenylation signal"
     CDS             complement(1982..2776)
                     /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(2093..2112)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2703..2722
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2808..2855)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(2870..3227)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2884..3019
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        3036..3227
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     3184..3204
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    3472..3593
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3509..3528)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3563..3582
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3630..3648
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     primer_bind     complement(5629..5648)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5630..5648)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5690..5709)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          5692..5824
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        complement(join(5960..6090,1..73))
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5989..6009)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 ggtggagact tggaaatccc cgtgagtcaa accgctatcc acgcccattg gtgtactgcc
       61 aaaaccgcat caccatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa
      121 agtcccgtaa ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt
      181 caataggggg cggacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc
      241 gtaaatactc cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat
      301 acgtcattat tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc gggccattta
      361 ccgtaagtta tgtaacgcgg aactccatat atgggctatg aactaatgac cccgtaattg
      421 attactatta ataactagtc aataatcaat gccaacatgg cggtcatatt ggacatgagc
      481 caatataaat gtacatatta tgatatagat acaacgtatg caatggccaa tagccaatat
      541 tgatttatgc tatataacca atgaataata tggctaatgg ccaatattga agatctcgat
      601 cctctacgcc ggacgcatcg tggccggagt actgcggttt atctcgatgg ctacgagggc
      661 agacagtaag tggatttacc ataatccctt aattgtacgc accgctaaaa cgcgttcagc
      721 gcgatcacgg cagcagacag gtaaaaatgg caacaaacca cccgaaaaac tgccgcaatc
      781 gcgcctgata aattttaacc gtatgaatac ctatgcaacc agagggtaca ggccacatta
      841 cccccactta atccactgaa gctgccattt ttcatggttt caccatccca gcgaagggcc
      901 atccagcgtg cgttcctgta tttccggact ataaaaggat ctaggtgaag atcctttttg
      961 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     1021 tagaaaagat caaaggatct tcttgaaatc ctttttttct gcgcgtaatc tgctgcttgc
     1081 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     1141 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     1201 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     1261 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     1321 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     1381 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     1441 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     1501 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     1561 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     1621 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     1681 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     1741 ttgagtgagc tgataccggg tagaaagcca gtccgcagaa acggtgctga ccccggatga
     1801 atgtcagcta ctgggctatc tggacaaggg aaaacgcaag cgcaaagagg atccttatcg
     1861 ctatcgattc acacaaaaaa ccaacacaca gatgtaatga aaataaagat attttattgc
     1921 ggccatcgtg atggcaggtt gggcgtcgct tggtcggtca tttcgaaccc cagagtcccg
     1981 ctcagaagaa ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga
     2041 taccgtaaag cacgaggaag cggtcagccc attcgccgcc aagctcttca gcaatatcac
     2101 gggtagccaa cgctatgtcc tgatagcggt ccgccacacc cagccggcca cagtcgatga
     2161 atccagaaaa gcggccattt tccaccatga tattcggcaa gcaggcatcg ccatgggtca
     2221 cgacgagatc ctcgccgtcg ggcatgcgcg ccttgagcct ggcgaacagt tcggctggcg
     2281 cgagcccctg atgctcttcg tccagatcat cctgatcgac aagaccggct tccatccgag
     2341 tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa
     2401 gcgtatgcag ccgccgcatt gcatcagcca tgatggatac tttctcggca ggagcaaggt
     2461 gagatgacag gagatcctgc cccggcactt cgcccaatag cagccagtcc cttcccgctt
     2521 cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc
     2581 gcgctgcctc gtcctgcagt tcattcaggg caccggacag gtcggtcttg acaaaaagaa
     2641 ccgggcgccc ctgcgctgac agccggaaca cggcggcatc agagcagccg attgtctgtt
     2701 gtgcccagtc atagccgaat agcctctcca cccaagcggc cggagaacct gcgtgcaatc
     2761 catcttgttc aatcatgatc tgcctcctgg gtttagttcc tcaccttgtc gtattatact
     2821 atgccgatat actatgccga tgattaattg tcaacagtta attaagcttt ttgcaaaagc
     2881 ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggcc gaggcggcct
     2941 cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg
     3001 gcggagttag gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga
     3061 gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca cacctggttg
     3121 ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc
     3181 cacaccctaa ctgacacaca ttccacagct ggttctttcc gcctcagaag gtacctaacc
     3241 aagttcctct ttcagaggtt atttcaggcc atggtgctgc gcagatccgc gtatgcggtg
     3301 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggaaattgt aagcgttaat
     3361 attttgttaa aattcgattt taccacattt gtagaggttt tacttgctta aaaaaacctc
     3421 ccacatctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     3481 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     3541 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     3601 tgctcgaagc ggccgcccca aggggttatg ctagttattg ctcagcggtg gcagcagcca
     3661 actcagcttc ctttcgggct ttgttagcag ccggatcagc ttgcatgcct gcaggtcgac
     3721 tctagagtcg cggccttagc taatcttctt gaacagccgc cagccgctca caccgctcga
     3781 gccacctgaa ctgccttgag aaactgaaag aaatgtacga actgtaccct tgattgtact
     3841 cctacaaata ggacactttc ttaaagaagg agcacaatct ttgcatacta ctagatgacc
     3901 acaaggaata aacactatgg acacttcttt gtccatacac actttacatg ttctttcttc
     3961 ttgtagtctc cgcaattgtt cttccactgg tagatctgaa acatcttctg tgggaatata
     4021 ttttatgtcc tgttgcacaa ataaatgctc atataacaca gcttcagctt cttgcagaga
     4081 gtttctgaat acagtggctg caatatttcc ttttactaaa atcgtatcaa tcagttctct
     4141 tgcttgtaaa gacgtctgtg tcttctgttt aataacatca tgttcttgtt cattaataat
     4201 tccggcagtt agtagactat ccaggattgg aattacacaa gtcaaatgtt gaaaaagtgc
     4261 cattctattc ttccggatta ataataaatc atttgattct ttttcctcag ttgctctttc
     4321 tctctcctct tcccttattt catcttctgc attgagtaag tctaacacaa gatcattgac
     4381 tagtctataa ttctctccag ttgctaggat ttttctctga actgtctgtt ttaccaggct
     4441 tctactaaag cccatttcca cggcagcatt aatcacagga gtattcatca tgattgcatc
     4501 ttctgaatgg tcttctccag gttcaaaatg gataattgat gactctgcat tttcatctcc
     4561 tgggctgtct gatgtggata gcagctgttc aagtagatga gggtaactgg cttgaacttg
     4621 acggatgaac tcctgtcctt taattcttat caagtactca caccttggaa accacttggc
     4681 atgttgaacc catggatcat ctccagattc ccaacacctg agtccaccat cacagcaaaa
     4741 gcatttgaca tcatcactgt tacccacata ataaaaaccc gcacttgcaa gctgctcagg
     4801 attaactaga acactagagg gccagttaaa gaatgtttta aagcgggctg catgtgtctg
     4861 catgctcaga ttagaaactg tgtatcttga agtgtcttga agctgatttt ctataaatgg
     4921 gcatttggga aaatgtctca ggtgttctga catagcatta tccttcggtt cccaattgct
     4981 caattttcca ccacaggcaa agcaagccac tctgtctcca ggtcctatgt agtaaaagcc
     5041 tgcttttgcc agatctgttg gcgacagaaa agtcaatggc catgtctgaa aagtaagtaa
     5101 tctggcattt tcgttattca ttgcacagtg gtaggaactt ctcatcaagg cagaaaaatc
     5161 ttgatttgct ctggagttta caggatttga tggagagttt gaataagagc cacggaaata
     5221 tccactgttt tctgtacccg gaagtaatga gtgtgtggaa tttgttactg aagaaggaaa
     5281 agtaggctga gaggtagctt ccaagttgtt aacggaattt agactctgaa cgaatctgca
     5341 gctaggatac aactttttat gcttttcagt aggactgtct cctcttttcc agttatccag
     5401 catcaggcca caacagaagc atttgacctt gtcattcaca ccagtgtaat agaaaccagc
     5461 acgagcaaga ctcctttctg agacaggaac cccagcagga aaagtggaat acgtagacat
     5521 tcggtacagt tcacatgaca agtcgtattt cagttcaaac gtgttggcgc ttttcatcaa
     5581 atttgataag aatatgctgt tttctactat gttcatggcg atcgctagcc ctatagtgag
     5641 tcgtattaat actctagcct taagagctgt aattgaactg ggagtggaca cctgtggaga
     5701 gaaaggcaaa gtggatgtca gtaagaccaa taggtgccta tcagaaacgc aagagtcttc
     5761 tctgtctcga caagcccagt ttctattggt ctccttaaac ctgtcttgta accttgatac
     5821 ttacctgccc agtgcctcac gaccaacttc tgcagcttaa gttcgagact gttgtgtcag
     5881 aagcactgac tgcgttagca atttaactgt gataaactac cgcaataaag cttctagtga
     5941 tctgacggtt cactaaacca gctctgctta tatagacctc ccaccgtaca cgcctaccgc
     6001 ccatttgcgt caacggggcg gggttattac gacattttgg aaagtcccgt tgattttggt
     6061 gccaaaacaa actcccattg acgtcaatgg
//
