LOCUS       sequence-486651-        2921 bp    DNA     circular SYN 17-SEP-2025
DEFINITION  Express NanoLuc(R)-NRAS-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  JOURNAL   Unpublished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 Sep 17, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: NanoLuc-NRAS-CRAF(RBD)-HaloTag BiBRET Vector
FEATURES             Location/Qualifiers
     source          1..2921
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..111
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(349..1143)
                     /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(460..479)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1070..1089
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1175..1222)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     protein_bind    1232..1265
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    1380..1461
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1417..1436)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1540..1897
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1540..1731
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1563..1583)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1748..1883
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      1922..1931
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1928..2326
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    2350..2398
                     /note="synthetic polyadenylation signal"
     rep_origin      join(2524..2921,1..191)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
       61 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
      121 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
      181 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
      241 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
      301 ttgagtgagc tgataccggt tgggcgtcgc ttggtcggtc atttcgaatc agaagaactc
      361 gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac
      421 gaggaagcgg tcagcccatt cgccgccaag ctcttcagca atatcacggg tagccaacgc
      481 tatgtcctga tagcggtccg ccacacccag ccggccacag tcgatgaatc cagaaaagcg
      541 gccattttcc accatgatat tcggcaagca ggcatcgcca tgggtcacga cgagatcctc
      601 gccgtcgggc atgcgcgcct tgagcctggc gaacagttcg gctggcgcga gcccctgatg
      661 ctcttcgtcc agatcatcct gatcgacaag accggcttcc atccgagtac gtgctcgctc
      721 gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg
      781 ccgcattgca tcagccatga tggatacttt ctcggcagga gcaaggtgag atgacaggag
      841 atcctgcccc ggcacttcgc ccaatagcag ccagtccctt cccgcttcag tgacaacgtc
      901 gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc
      961 ctgcagttca ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg
     1021 cgctgacagc cggaacacgg cggcatcaga gcagccgatt gtctgttgtg cccagtcata
     1081 gccgaatagc ctctccaccc aagcggccgg agaacctgcg tgcaatccat cttgttcaat
     1141 catgatctgc ctcctgggtt tagttcctca ccttgtcgta ttatactatg ccgatatact
     1201 atgccgatga ttaattgtca acagtggatc cataacttcg tataatgtat gctatacgaa
     1261 gttatattag aattcttaat taagatttta ccacatttgt agaggtttta cttgcttaaa
     1321 aaaacctccc acatctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     1381 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     1441 ataaagcatt tttttcactg ccgacgtctt aattaacgcg tcttgcgcag caccatggcc
     1501 tgaaataacc tctgaaagag gaacttggtt aggtaccttc tgaggcggaa agaaccagct
     1561 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     1621 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     1681 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     1741 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     1801 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     1861 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcgattctt ctgacactag
     1921 cgccaccatg gccaagcctt tgtctcaaga agaatccacc ctcattgaaa gagcaacggc
     1981 tacaatcaac agcatcccca tctctgaaga ctacagcgtc gccagcgcag ctctctctag
     2041 cgacggccgc atcttcactg gtgtcaatgt atatcatttt actgggggac cttgtgcaga
     2101 actcgtggtg ctgggcactg ctgctgctgc ggcagctggc aacctgactt gtatcgtcgc
     2161 gatcggaaat gagaacaggg gcatcttgag cccctgcgga cggtgccgac aggtgcttct
     2221 cgatctgcat cctgggatca aagccatagt gaaggacagt gatggacagc cgacggcagt
     2281 tgggattcgt gaattgctgc cctctggtta tgtgtgggag ggctaaactc tagccatcac
     2341 gatggccgca ataaaatatc tttattttca ttacatctgt gtgttggttt tttgtgtgac
     2401 gattccggac tataaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     2461 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     2521 ttcttgaaat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     2581 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     2641 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     2701 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     2761 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     2821 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     2881 gacctacacc gaactgagat acctacagcg tgagctatga g
//
