LOCUS       sequence-336180-        6597 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of myc-GBAi W211A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6597)
  AUTHORS   Leyme A, Marivin A, Maziarz M, DiGiacomo V, Papakonstantinou MP, 
            Patel PP, Blanco-Canosa JB, Walawalkar IA, Rodriguez-Davila G, 
            Dominguez I, Garcia-Marcos M
  TITLE     Specific inhibition of GPCR-independent G protein signaling by a 
            rationally engineered protein.
  JOURNAL   Proc Natl Acad Sci U S A. 2017 Nov 28;114(48):E10319-E10328. doi: 
            10.1073/pnas.1707992114. Epub 2017 Nov 13.
  PUBMED    29133411
REFERENCE   2  (bases 1 to 6597)
  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"; doi: 
            "10.1073/pnas.1707992114"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2017-11-28- 28"; volume: "114"; issue: "48"; pages: 
            "E10319-E10328"
COMMENT     Sequence Label: pLIC-myc-GBAi W211A
FEATURES             Location/Qualifiers
     source          1..6597
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             704..733
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(1785..1802)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1791..2015
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2061..2489
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2148..2167)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2358..2379
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2498..2518)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2503..2832
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2683..2818
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2745..2764
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2881..3906
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             2881..3906
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    4036..4157
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4073..4092)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4127..4146
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4206..4222)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4206..4222)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4219..4241)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4230..4246
                     /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(4254..4260,4261..4278,4279..4284))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4299..4320
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4437..4454)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4608..5196)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4688..4707)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5367..6158,6159..6227))
                     /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     5990..6009
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6228..6332)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6407..6426)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacttaagc ttggtaccga gctcggatcc atggaacaaa aacttatttc
      721 tgaagaagat ctgggaattc tgtacttcca atccaatgcg gacggagaga aagcggccaa
      781 agaagtgaag ctgctgctgc tcggcgctgg agaatctggt aaaagtacta ttgtgaaaca
      841 gatgaaaatc attcatgagg atggctattc cgaggacgag tgtaagcagt ataaagttgt
      901 cgtctacagc aataccattc agtccatcat tgcaatcata agagccatgg gacggttgaa
      961 gattgatttt ggggaagctg ccagagcgga tgatgcccga cagttatttg ttttagctgg
     1021 cagtgctgag gaaggagtca tgacttcaga actagcaggc gtgattaaac gtttatggcg
     1081 agatggcggg gtgcaggcat gcttcagcag gtccagggaa tatcagctca ttgattctgc
     1141 ttcatattac ctaaatgatt tggatagaat atcccagacc aactacattc caactcagca
     1201 agatgttctt cggacgagag tgaagactac aggcattgtg gagacccact tcaccttcaa
     1261 ggagctctac ttcaaaatgt ttgatgtagg tgcccaaaga tccgaacgga aaaaggcgat
     1321 ccactgtttt gagggagtga cagcaattat cttttgtgtg gctctcagtg attacgacct
     1381 tgttctggct gaggacgagg aaatgaaccg aatgcatgaa agcatgaaat tgtttgacag
     1441 catttgtaac aacaaatggt ttacagacac ttcaatcatt ctcttcctta ataagaaaga
     1501 cctttttgag gaaaaaataa agaggagtcc attaacaatc tgttatccag aatacacagg
     1561 ttccaatacg tacgaagagg cagctgctta catccagtgc cagtttgaag atctgaaccg
     1621 aagaaaggac accaaggagg tctacactca ctttacctgt gccactgaca ccaaaaacgt
     1681 gcagtttgtt tttgatgctg ttacagatgt catcattaaa tagcgcattg gaagtggata
     1741 actcgagcat gcatctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
     1801 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     1861 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     1921 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     1981 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     2041 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2101 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     2161 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     2221 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     2281 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     2341 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     2401 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     2461 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     2521 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     2581 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     2641 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     2701 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     2761 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     2821 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg
     2881 atgaaaaagc ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac
     2941 agcgtctccg acctgatgca gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat
     3001 gtaggagggc gtggatatgt cctgcgggta aatagctgcg ccgatggttt ctacaaagat
     3061 cgttatgttt atcggcactt tgcatcggcc gcgctcccga ttccggaagt gcttgacatt
     3121 ggggaattta gcgagagcct gacctattgc atctcccgcc gtgcacaggg tgtcacgttg
     3181 caagacctgc ctgaaaccga actgcccgct gttctgcagc cggtcgcgga ggccatggat
     3241 gcgatcgctg cggccgatct tagccagacg agcgggttcg gcccattcgg accgcaagga
     3301 atcggtcaat acactacatg gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat
     3361 cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag
     3421 ctgatgcttt gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc
     3481 tccaacaatg tcctgacgga caatggccgc ataacagcgg tcattgactg gagcgaggcg
     3541 atgttcgggg attcccaata cgaggtcgcc aacatcttct tctggaggcc gtggttggct
     3601 tgtatggagc agcagacgcg ctacttcgag cggaggcatc cggagcttgc aggatcgccg
     3661 cggctccggg cgtatatgct ccgcattggt cttgaccaac tctatcagag cttggttgac
     3721 ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga
     3781 gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc
     3841 tgtgtagaag tactcgccga tagtggaaac cgacgcccca gcactcgtcc gagggcaaag
     3901 gaatagcacg tgctacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     3961 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     4021 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     4081 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     4141 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     4201 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     4261 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     4321 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     4381 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     4441 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4501 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4561 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4621 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4681 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4741 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4801 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4861 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4921 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4981 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5041 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5101 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5161 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5221 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5281 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5341 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     5401 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     5461 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     5521 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     5581 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     5641 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     5701 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     5761 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     5821 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     5881 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     5941 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6001 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6061 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6121 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     6181 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     6241 caatcttatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     6301 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     6361 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
     6421 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
     6481 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
     6541 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgtt
//
