LOCUS       sequence-472950-        2921 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express NanoLuc(R)-HRAS WT-CRAF RBD-HaloTag(R) in Mammalian Cells 
            under a CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2921)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 2921)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: NanoBRET BiBRET Vector, NanoLuc-HRAS WT-CRAF
            R...
FEATURES             Location/Qualifiers
     source          1..2921
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    398..446
                     /note="synthetic polyadenylation signal"
     CDS             complement(470..868)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      865..874
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(899..1256)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      913..1048
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        1065..1256
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     1213..1233
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    1335..1416
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     1360..1379
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     protein_bind    complement(1531..1564)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        1574..1621
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1653..2447
                     /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(1707..1726)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2317..2336
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(join(2605..2921,1..272))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2685..2704)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
       61 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
      121 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
      181 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
      241 agcagattac gcgcagaaaa aaaggatttc aagaagatcc tttgatcttt tctacggggt
      301 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
      361 ggatcttcac ctagatcctt ttatagtccg gaatcgtcac acaaaaaacc aacacacaga
      421 tgtaatgaaa ataaagatat tttattgcgg ccatcgtgat ggctagagtt tagccctccc
      481 acacataacc agagggcagc aattcacgaa tcccaactgc cgtcggctgt ccatcactgt
      541 ccttcactat ggctttgatc ccaggatgca gatcgagaag cacctgtcgg caccgtccgc
      601 aggggctcaa gatgcccctg ttctcatttc cgatcgcgac gatacaagtc aggttgccag
      661 ctgccgcagc agcagcagtg cccagcacca cgagttctgc acaaggtccc ccagtaaaat
      721 gatatacatt gacaccagtg aagatgcggc cgtcgctaga gagagctgcg ctggcgacgc
      781 tgtagtcttc agagatgggg atgctgttga ttgtagccgt tgctctttca atgagggtgg
      841 attcttcttg agacaaaggc ttggccatgg tggcgctagt gtcagaagaa tcgagctttt
      901 tgcaaaagcc taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggccg
      961 aggcggcctc ggcctctgca taaataaaaa aaattagtca gccatggggc ggagaatggg
     1021 cggaactggg cggagttagg ggcgggatgg gcggagttag gggcgggact atggttgctg
     1081 actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac
     1141 acctggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg
     1201 gggactttcc acaccctaac tgacacacat tccacagctg gttctttccg cctcagaagg
     1261 tacctaacca agttcctctt tcagaggtta tttcaggcca tggtgctgcg caagacgcgt
     1321 taattaagac gtcggcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     1381 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     1441 atgtttcagg ttcaggggga gatgtgggag gtttttttaa gcaagtaaaa cctctacaaa
     1501 tgtggtaaaa tcttaattaa gaattctaat ataacttcgt atagcataca ttatacgaag
     1561 ttatggatcc actgttgaca attaatcatc ggcatagtat atcggcatag tataatacga
     1621 caaggtgagg aactaaaccc aggaggcaga tcatgattga acaagatgga ttgcacgcag
     1681 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     1741 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     1801 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     1861 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     1921 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     1981 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     2041 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     2101 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     2161 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     2221 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     2281 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     2341 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     2401 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgattc gaaatgaccg
     2461 accaagcgac gcccaaccgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     2521 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     2581 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     2641 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     2701 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     2761 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     2821 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     2881 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc a
//
