LOCUS       sequence-198178-        8037 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human R-Ras2 with FLAG tag for mammalian expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8037)
  AUTHORS   Zhang X, Spiegelman NA, Nelson OD, Jing H, Lin H
  TITLE     SIRT6 regulates Ras-related protein R-Ras2 by lysine 
            defatty-acylation.
  JOURNAL   Elife. 2017 Apr 13;6. pii: e25158. doi: 10.7554/eLife.25158.
  PUBMED    28406396
REFERENCE   2  (bases 1 to 8037)
  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: "Elife."; 
            date: "2017-04-13"; volume: "6. pii: e25158. doi"; pages: " 
            10.7554/eLife.25158"
COMMENT     Sequence Label: pCDH-FLAG-R-Ras2
FEATURES             Location/Qualifiers
     source          1..8037
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        9..212
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     163..183
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     209..233
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             298..321
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        987..1198
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             1211..1479
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             1506..2105
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1506..1525)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2002..2022
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    2112..2700
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2165..2185)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2583..2594)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2774..3007
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3079..3200
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3116..3135)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3170..3189
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3240..3375
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3408..3424)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3408..3424)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3421..3443)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3432..3448
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3456..3462,3463..3480,3481..3486))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3501..3522
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3639..3656)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3810..4398)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(4569..5360,5361..5429))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5192..5211
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5430..5534)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5602..5620
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5658..5680)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5780..5799
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5993..6015
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6007..6024
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6008..6024
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6039..6265
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6266..6446
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6493..6618
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7111..7344
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7529..7573
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    7840..7956
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
       61 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      121 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
      181 gtgtacggtg ggaggtttat ataagcagag ctcgtttagt gaaccgtcag atcgcctgga
      241 gacgccatcc acgctgtttt gacctccata gaagattcta gagctagcga attcatggat
      301 tacaaggatg acgacgataa gggaatggcc gcggccggct ggcgggacgg ctccggccag
      361 gagaagtacc ggctcgtggt ggtcggcggg ggcggcgtgg gcaagtcggc gctcaccatc
      421 cagttcatcc agtcctattt tgtaacggat tatgatccaa ccattgaaga ttcttacaca
      481 aagcagtgtg tgatagatga cagagcagcc cggctagata ttttggatac agcaggacaa
      541 gaagagtttg gagccatgag agaacagtat atgaggactg gcgaaggctt cctgttggtc
      601 ttttcagtca cagatagagg cagttttgaa gaaatctata agtttcaaag acagattctc
      661 agagtaaagg atcgtgatga gttcccaatg attttaattg gtaataaagc agatctggat
      721 catcaaagac aggtaacaca ggaagaagga caacagttag cacggcagct taaggtaaca
      781 tacatggagg catcagcaaa gattaggatg aatgtagatc aagctttcca tgaacttgtc
      841 cgggttatca ggaaatttca agagcaggaa tgtcctcctt caccagaacc aacacggaaa
      901 gaaaaagaca agaaaggctg ccattgtgtc attttctagg gatccgcggc cgcgaaggat
      961 ctgcgatcgc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa
     1021 gttgggggga ggggtcggca attgaacggg tgcctagaga aggtggcgcg gggtaaactg
     1081 ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat
     1141 aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacagct
     1201 gaagcttcga ggggctcgca tctctccttc acgcgcccgc cgccctacct gaggccgcca
     1261 tccacgccgg ttgagtcgcg ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc
     1321 cgccgtctag gtaagtttaa agctcaggtc gagaccgggc ctttgtccgg cgctcccttg
     1381 gagcctacct agactcagcc ggctctccac gctttgcctg accctgcttg ctcaactcta
     1441 cgtctttgtt tcgttttctg ttctgcgccg ttacagatcc aagctgtgac cggcgcctac
     1501 gctagatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg
     1561 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc
     1621 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc
     1681 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc
     1741 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga
     1801 gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca
     1861 aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc
     1921 tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct
     1981 tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca
     2041 ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg
     2101 cctgagtcga caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     2161 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     2221 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     2281 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     2341 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     2401 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     2461 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc
     2521 cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     2581 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     2641 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     2701 ctggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa
     2761 gaaaaggggg gactggaagg gctaattcac tcccaacgaa gataagatct gctttttgct
     2821 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     2881 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     2941 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     3001 tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga
     3061 atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     3121 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     3181 aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca
     3241 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3301 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     3361 gcttttttgg aggcctagac ttttgcagag accaaattcg taatcatgtc atagctgttt
     3421 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     3481 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     3541 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     3601 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     3661 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     3721 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     3781 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3841 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3901 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     3961 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     4021 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4081 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4141 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4201 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     4261 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     4321 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     4381 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     4441 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     4501 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     4561 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     4621 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     4681 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     4741 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     4801 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     4861 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     4921 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     4981 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     5041 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     5101 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     5161 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     5221 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     5281 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     5341 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     5401 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     5461 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     5521 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     5581 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc
     5641 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     5701 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     5761 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg
     5821 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc
     5881 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     5941 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     6001 cgacgttgta aaacgacggc cagtgccaag ctgacgcgtg tagtcttatg caatactctt
     6061 gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc ttacaaggag agaaaaagca
     6121 ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt gccttattag gaaggcaaca
     6181 gacgggtctg acatggattg gacgaaccac tgaattgccg cattgcagag atattgtatt
     6241 taagtgccta gctcgataca taaacgggtc tctctggtta gaccagatct gagcctggga
     6301 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
     6361 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
     6421 ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg
     6481 aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac ggcaagaggc
     6541 gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta gaaggagaga
     6601 gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt
     6661 cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg ggcaagcagg
     6721 gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg ctgtagacaa
     6781 atactgggac agctacaacc atcccttcag acaggatcag aagaacttag atcattatat
     6841 aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga caccaaggaa
     6901 gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc
     6961 actgatcttc agacctggag gaggagatat gagggacaat tggagaagtg aattatataa
     7021 atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa agagaagagt
     7081 ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt tcttgggagc
     7141 agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca gacaattatt
     7201 gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc aacagcatct
     7261 gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg ctgtggaaag
     7321 atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac tcatttgcac
     7381 cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga tttggaatca
     7441 cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa tacactcctt
     7501 aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg aattagataa
     7561 atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata taaaattatt
     7621 cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac tttctatagt
     7681 gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc caaccccgag
     7741 gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca gagacagatc
     7801 cattcgatta gtgaacggat ctcgacggta tcggttaact tttaaaagaa aaggggggat
     7861 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     7921 agaattacaa aaacaaatta caaaattcaa aattttatcg atactagtat tatgcccagt
     7981 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgcta
//
