LOCUS       sequence-215433-        8014 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mutant RGS7- E383K- in mamalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8014)
  AUTHORS   Qutob N, Masuho I, Alon M, Emmanuel R, Cohen I, Di Pizio A, Madore 
            J, Elkahloun A, Ziv T, Levy R, Gartner JJ, Hill VK, Lin JC, Hevroni 
            Y, Greenberg P, Brodezki A, Rosenberg SA, Kosloff M, Hayward NK, 
            Admon A, Niv MY, Scolyer RA, Martemyanov KA, Samuels Y
  TITLE     RGS7 is recurrently mutated in melanoma and promotes migration and 
            invasion of human cancer cells.
  JOURNAL   Sci Rep. 2018 Jan 12;8(1):653. doi: 10.1038/s41598-017-18851-4.
  PUBMED    29330521
REFERENCE   2  (bases 1 to 8014)
  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: "Sci Rep."; 
            date: "2018-01-12"; volume: "8(1):653. doi"; pages: " 
            10.1038/s41598-017-18851-4"
COMMENT     Sequence Label: pCDF1-MCS2-EF1-Puro-RGS7-E383K
FEATURES             Location/Qualifiers
     source          1..8014
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        133..336
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        1479..1682
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1679..1703
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3179..3202
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        3247..3458
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             3471..3739
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             3766..4365
                     /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(3766..3785)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4262..4282
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    4372..4960
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4425..4445)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4843..4854)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5298..5419
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5335..5354)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5389..5408
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5438..5573
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5611..5627)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5611..5627)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5624..5646)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5635..5651
                     /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(5659..5665,5666..5683,5684..5689))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5704..5725
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5842..5859)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6013..6601)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6093..6112)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6772..7563,7564..7632))
                     /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     7395..7414
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7633..7737)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7805..7823
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7861..7883)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7983..8002
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
       61 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      121 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      181 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      241 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
      301 aggcgtgtac ggtgggaggt ctatataagc agagcttgtg aaacttcgag gagtctcttt
      361 gttgaggact tttgagttct cccttgaggc tcccacagat acaataaata tttgagattg
      421 aaccctgtcg agtatctgtg taatcttttt tacctgtgag gtctcggaat ccgggccgag
      481 aacttcgcag ttggcgcccg aacagggact tgattgagag tgattgagga agtgaagcta
      541 gagcaataga aagctgttaa gcagaactcc tgctgaccta aatagggaag cagtagcaga
      601 cgctgctaac agtgagtatc tctagtgaag cagactcgag ctcataatca agtcattgtt
      661 taaaggccca gataaattac atctggtgac tcttcgcgga ccttcaagcc aggagattcg
      721 ccgagggaca gtcaacaagg taggagagat tctacagcaa catggggaat ggacaggggc
      781 gagattggaa aatggccatt aagagatgta gtaatgttgc tgtaggagta ggggggaaga
      841 gtaaaaaatt tggagaaggg aatttcagat gggccattag aatggctaat gtatctacag
      901 gacgagaacc tggtgatata ccagagactt tagatcaact aaggttggtt atttgcgatt
      961 tacaagaaag aagagaaaaa tttggatcta gcaaagaaat tgatatggca attcctgcat
     1021 tgaggagaaa tggtaggcaa tgtggcatgt ctgaaaaaga ggaggaatga tgaagtatct
     1081 cagacttatt ttataaggga gatactgtgc tgagttcttc cctttgagga aggtatgtca
     1141 tatcctagac atagtctcaa ttttaaaaga agaggtagga taggagggat ggccccttat
     1201 gaattattag cacaacaaga atccttaaga atacaagatt atttttctgc aataccacaa
     1261 aaattgcaag cacagtggat ttattataaa gatcaaaaag ataagaaatg gaaaggacca
     1321 atgagagtag aatactgggg acagggatca gtattattaa aggatgaaga gaagggatat
     1381 tttcttataa tcgatactag tattatgccc agtacatgac cttatgggac tttcctactt
     1441 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
     1501 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     1561 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     1621 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     1681 ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata
     1741 gaagattcta gagcccgggc gcgccggatc catggcccag gggaataatt atgggcagac
     1801 cagcaacggg gtggccgatg aatcacccaa catgctggtg tacagaaaga tggaagacgt
     1861 catagcacgg atgcaagatg aaaaaaatgg aattcctatt cgtacggtca aaagctttct
     1921 ttccaagata cctagcgtct tctctggttc agacattgtt caatggttga taaagaactt
     1981 aactatagaa gatccagtgg aggcgctcca tttgggaaca ttaatggctg cccacggcta
     2041 cttctttcca atctcagatc atgtcctcac actcaaggat gatggcacct tttaccggtt
     2101 tcaaaccccc tatttttggc catcaaattg ttgggagccg gaaaacacag attatgccgt
     2161 ttacctctgc aagagaacaa tgcaaaacaa ggcacgactg gagctcgcag actatgaggc
     2221 tgagagcctg gccaggctgc agagagcatt tgcccggaag tgggagttca ttttcatgca
     2281 agcagaagca caagcaaaag tggacaagaa gagagacaag attgaaagga agatccttga
     2341 cagccaagag agagcgttct gggacgtgca caggcccgtg cctggatgtg taaatacaac
     2401 tgaagtggac attaagaagt catccagaat gagaaacccc cacaaaacac ggaagtctgt
     2461 ctatggttta caaaatgata ttagaagtca cagtcctacc cacacaccca caccagaaac
     2521 taaacctcca acagaagatg agttacaaca acagataaaa tattggcaaa tacagttaga
     2581 tagacatcgg ttaaaaatgt caaaagtcgc tgacagtcta ctaagttaca cggaacagta
     2641 tttagaatac gacccgtttc ttttgccacc tgacccttct aacccatggc tgtccgatga
     2701 caccactttc tgggaacttg aggcaagcaa agaaccgagc cagcagaggg taaaacgatg
     2761 gggttttggc atggacgagg cattgaaaga cccagttggg agagaacagt tccttaaatt
     2821 tctagagtca gaattcagct cggaaaattt aagattctgg ctggcagtgg aggacctgaa
     2881 aaagaggcct attaaagaag taccctcaag agttcagaaa atatggcaag agtttctggc
     2941 tcccggagcc cccagtgcta ttaacttgga ttccaagagt tatgacaaaa ccacacagaa
     3001 cgtgaaggaa cctggacgat acacatttga agatgctcag gagcacattt acaaactgat
     3061 gaaaagtgat tcatacccac gttttataag atccagtgcc tatcaggagc ttctacaggc
     3121 aaagaaaaag gggaaatctc tcacgtccaa gaggttaaca agccttgctc agtcttacga
     3181 ttacaaggat gacgacgata agtaagcggc cgcgaaggat ctgcgatcgc tccggtgccc
     3241 gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca
     3301 attgaacggg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact
     3361 ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga
     3421 acgttctttt tcgcaacggg tttgccgcca gaacacagct gaagcttcga ggggctcgca
     3481 tctctccttc acgcgcccgc cgccctacct gaggccgcca tccacgccgg ttgagtcgcg
     3541 ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa
     3601 agctcaggtc gagaccgggc ctttgtccgg cgctcccttg gagcctacct agactcagcc
     3661 ggctctccac gctttgcctg accctgcttg ctcaactcta cgtctttgtt tcgttttctg
     3721 ttctgcgccg ttacagatcc aagctgtgac cggcgcctac gctagatgac cgagtacaag
     3781 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc
     3841 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
     3901 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     3961 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     4021 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     4081 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     4141 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     4201 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     4261 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     4321 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgagtcga caatcaacct
     4381 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     4441 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     4501 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     4561 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     4621 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     4681 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     4741 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt
     4801 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     4861 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     4921 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctgggtaccg atgacagagt
     4981 tagaagatcg cttcaggaag ctatttggca cgacttctac aacgggagac agcacagtag
     5041 attctgaaga tgaacctcct aaaaaagaaa aaagggtgga ctgggatgag tattggaacc
     5101 ctgaaatcga tagcttccag tgctttgtga aacttcgagg agtctctttg ttgaggactt
     5161 ttgagttctc ccttgaggct cccacagata caataaatat ttgagattga accctgtcga
     5221 gtatctgtgt aatctttttt acctgtgagg tctcggaatc cgggccgaga acttcgcagc
     5281 gagctcattg taccgcgaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     5341 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     5401 aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccag
     5461 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     5521 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagactt
     5581 ttgcagagac ggcccaaatt cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     5641 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     5701 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     5761 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     5821 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     5881 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5941 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     6001 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     6061 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     6121 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     6181 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     6241 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     6301 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6361 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6421 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     6481 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6541 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     6601 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6661 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     6721 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     6781 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     6841 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     6901 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     6961 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     7021 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     7081 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     7141 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     7201 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     7261 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     7321 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     7381 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     7441 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     7501 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     7561 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     7621 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     7681 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     7741 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     7801 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     7861 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     7921 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     7981 tgcggcatca gagcagattg tactgagagt gcac
//
