LOCUS       sequence-413191-        4506 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of HRas with G12V mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4506)
  AUTHORS   Sekhon H, Ha JH, Presti MF, Procopio SB, Jarvis AR, Mirsky PO, John 
            AM, Loh SN
  TITLE     Adaptable, turn-on maturation (ATOM) fluorescent biosensors for 
            multiplexed detection in cells.
  JOURNAL   Nat Methods. 2023 Nov 9. doi: 10.1038/s41592-023-02065-w.
  PUBMED    37945909
REFERENCE   2  (bases 1 to 4506)
  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: "Nat 
            Methods. 2023 Nov 9. doi: 10.1038/s41592-023-02065-w."
COMMENT     Sequence Label: pCMV-HRas(G12V)
FEATURES             Location/Qualifiers
     source          1..4506
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    609..665
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    1294..1415
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1331..1350)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1385..1404
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1422..1877)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1559..1580)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1771..1790
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1904..2008
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2010..2367
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2010..2201
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2033..2053)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2218..2353
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2402..3196
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2402..3196
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2456..2475)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3066..3085
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3384..3403)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3428..3475
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3804..4392
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4293..4312
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc gcgggcccgg
      661 gatccatgac agaatacaag ttagtagtcg tcggtgctgt tggtgttggt aaatccgctt
      721 tgacaataca attaatacaa aatcacttcg tagatgaata cgatccaact atcgaagatt
      781 catacagaaa gcaagttgtt attgatggtg aaacttgttt gttggatatc ttggatacag
      841 ctggtcaaga agaatattct gcaatgagag atcaatacat gagaactggt gaaggtttct
      901 tgtgtgtttt tgctattaat aacacaaaat catttgaaga tattcatcaa tatagagaac
      961 aaattaaaag agttaaagat tctgatgatg ttccaatggt tttggttggt aataagtgtg
     1021 atttggctgc aagaactgtt gaatctagac aagctcaaga tttggcaaga tcttatggta
     1081 ttccatacat tgaaacttct gctaaaacaa gacaaggtgt tgaagatgca ttctacacat
     1141 tggttagaga aatcagacaa catggtggat aagcggccgc gactctagat cataatcagc
     1201 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     1261 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     1321 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     1381 agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat
     1441 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     1501 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     1561 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     1621 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     1681 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     1741 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     1801 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     1861 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     1921 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     1981 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
     2041 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     2101 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     2161 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     2221 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     2281 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     2341 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
     2401 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     2461 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     2521 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     2581 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     2641 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     2701 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     2761 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     2821 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     2881 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
     2941 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     3001 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     3061 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     3121 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     3181 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     3241 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     3301 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     3361 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     3421 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
     3481 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     3541 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     3601 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata
     3661 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     3721 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     3781 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     3841 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     3901 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     3961 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     4021 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     4081 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     4141 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     4201 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     4261 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     4321 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     4381 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     4441 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     4501 atgcat
//
