LOCUS       sequence-338020-        6597 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of myc-GBAi.
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
FEATURES             Location/Qualifiers
     source          1..6597
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(84..104)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        89..418
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      269..404
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     331..350
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             467..1492
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             467..1492
                     /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    1622..1743
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1659..1678)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1713..1732
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1792..1808)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1792..1808)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1805..1827)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1816..1832
                     /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(1840..1846,1847..1864,1865..1870))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1885..1906
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2023..2040)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2194..2782)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2274..2293)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2953..3744,3745..3813))
                     /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     3576..3595
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3814..3918)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3993..4012)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4184..4563
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4564..4767
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4718..4738
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4812..4831
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4812..4830
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4887..4916
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(5968..5985)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5974..6198
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(6244..6597,1..75)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6331..6350)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6541..6562
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
       61 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
      121 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
      181 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
      241 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
      301 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
      361 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
      421 gctcccggga gcttgtatat ccattttcgg atctgatcag cacgtgatga aaaagcctga
      481 actcaccgcg acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg tctccgacct
      541 gatgcagctc tcggagggcg aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg
      601 atatgtcctg cgggtaaata gctgcgccga tggtttctac aaagatcgtt atgtttatcg
      661 gcactttgca tcggccgcgc tcccgattcc ggaagtgctt gacattgggg aatttagcga
      721 gagcctgacc tattgcatct cccgccgtgc acagggtgtc acgttgcaag acctgcctga
      781 aaccgaactg cccgctgttc tgcagccggt cgcggaggcc atggatgcga tcgctgcggc
      841 cgatcttagc cagacgagcg ggttcggccc attcggaccg caaggaatcg gtcaatacac
      901 tacatggcgt gatttcatat gcgcgattgc tgatccccat gtgtatcact ggcaaactgt
      961 gatggacgac accgtcagtg cgtccgtcgc gcaggctctc gatgagctga tgctttgggc
     1021 cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca acaatgtcct
     1081 gacggacaat ggccgcataa cagcggtcat tgactggagc gaggcgatgt tcggggattc
     1141 ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta tggagcagca
     1201 gacgcgctac ttcgagcgga ggcatccgga gcttgcagga tcgccgcggc tccgggcgta
     1261 tatgctccgc attggtcttg accaactcta tcagagcttg gttgacggca atttcgatga
     1321 tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga tccggagccg ggactgtcgg
     1381 gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc gatggctgtg tagaagtact
     1441 cgccgatagt ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat agcacgtgct
     1501 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     1561 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     1621 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     1681 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     1741 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     1801 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     1861 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     1921 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     1981 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     2041 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     2101 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     2161 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     2221 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     2281 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     2341 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     2401 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     2461 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     2521 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     2581 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     2641 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     2701 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     2761 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     2821 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     2881 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     2941 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     3001 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     3061 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     3121 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     3181 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     3241 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     3301 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     3361 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     3421 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     3481 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     3541 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     3601 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     3661 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     3721 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     3781 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat cttattgaag
     3841 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     3901 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     3961 agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     4021 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
     4081 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg
     4141 cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact
     4201 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     4261 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     4321 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     4381 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     4441 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     4501 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     4561 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
     4621 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
     4681 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
     4741 cggtgggagg tctatataag cagagctctc tggctaacta gagaacccac tgcttactgg
     4801 cttatcgaaa ttaatacgac tcactatagg gagacccaag ctggctagcg tttaaactta
     4861 agcttggtac cgagctcgga tccatggaac aaaaacttat ttctgaagaa gatctgggaa
     4921 ttctgtactt ccaatccaat gcggacggag agaaagcggc caaagaagtg aagctgctgc
     4981 tgctcggcgc tggagaatct ggtaaaagta ctattgtgaa acagatgaaa atcattcatg
     5041 aggatggcta ttccgaggac gagtgtaagc agtataaagt tgtcgtctac agcaatacca
     5101 ttcagtccat cattgcaatc ataagagcca tgggacggtt gaagattgat tttggggaag
     5161 ctgccagagc ggatgatgcc cgacagttat ttgttttagc tggcagtgct gaggaaggag
     5221 tcatgacttc agaactagca ggcgtgatta aacgtttatg gcgagatggc ggggtgcagg
     5281 catgcttcag caggtccagg gaatatcagc tcattgattc tgcttcatat tacctaaatg
     5341 atttggatag aatatcccag accaactaca ttccaactca gcaagatgtt cttcggacga
     5401 gagtgaagac tacaggcatt gtggagaccc acttcacctt caaggagctc tacttcaaaa
     5461 tgtttgatgt aggtgcccaa agatccgaac ggaaaaagtg gatccactgt tttgagggag
     5521 tgacagcaat tatcttttgt gtggctctca gtgattacga ccttgttctg gctgaggacg
     5581 aggaaatgaa ccgaatgcat gaaagcatga aattgtttga cagcatttgt aacaacaaat
     5641 ggtttacaga cacttcaatc attctcttcc ttaataagaa agaccttttt gaggaaaaaa
     5701 taaagaggag tccattaaca atctgttatc cagaatacac aggttccaat acgtacgaag
     5761 aggcagctgc ttacatccag tgccagtttg aagatctgaa ccgaagaaag gacaccaagg
     5821 aggtctacac tcactttacc tgtgccactg acaccaaaaa cgtgcagttt gtttttgatg
     5881 ctgttacaga tgtcatcatt aaatagcgca ttggaagtgg ataactcgag catgcatcta
     5941 gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     6001 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     6061 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     6121 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     6181 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc
     6241 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     6301 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     6361 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     6421 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     6481 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     6541 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttg
//
