LOCUS       sequence-112177-        6387 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In addition to the mutations in alpha-s(alpha3beta5), this dominant 
            negative G protein alpha-s mutant contains the G226A mutation, which
            impairs activating conformation changes in Switch II..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6387)
  AUTHORS   Berlot CH
  TITLE     A highly effective dominant negative alpha s construct containing 
            mutations that affect distinct functions inhibits multiple 
            Gs-coupled receptor signaling pathways.
  JOURNAL   J Biol Chem. 2002 Jun 7;277(23):21080-5. Epub 2002 Apr 1.
  PUBMED    11927592
REFERENCE   2  (bases 1 to 6387)
  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: "J Biol 
            Chem."; date: "2002-06-7"; volume: "277(23)"; pages: "21080-5. Epub 
            2002 Apr 1"
COMMENT     Sequence Label: G protein alpha-s(alpha3beta5/G226A)
FEATURES             Location/Qualifiers
     source          1..6387
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     471..490
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     rep_origin      complement(603..1061)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(740..761)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     955..974
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(1186..1204)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1272..1376
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1377..1445,1446..2237)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1595..1614)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     enhancer        2310..2689
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2690..2893
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2844..2864
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2938..2957
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2938..2956
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3160..3169
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4659..4677)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4660..4677)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     intron          4761..4826
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             4956..4976
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    5248..5382
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5285..5304)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5339..5358
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6228..6363
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 ggcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
       61 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
      121 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
      181 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
      241 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
      301 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
      361 ctacagcgtg agcattgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
      421 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
      481 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
      541 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcaagcta gcttctagct
      601 agaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
      661 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagcccga
      721 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      781 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc cgcccactac gtgaaccatc
      841 acccaaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
      901 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
      961 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     1021 caccacaccc gccgcgctta atgcgccgct acagggcgcg tactatggtt gctttgacga
     1081 gaccgtataa cgtgctttcc tcgttggaat cagagcggga gctaaacagg aggccgatta
     1141 aagggatttt agacaggaac ggtacgccag ctggattacc aaagggcctc gtgatacgcc
     1201 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     1261 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     1321 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     1381 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     1441 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     1501 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     1561 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg
     1621 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     1681 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     1741 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     1801 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     1861 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     1921 cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     1981 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     2041 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     2101 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     2161 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     2221 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     2281 aacttcattt ttaatttctc tagcgcgttg acattgatta ttgactagtt attaatagta
     2341 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     2401 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     2461 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggactattt
     2521 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     2581 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     2641 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     2701 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     2761 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     2821 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     2881 tataagcaga gctctctggc taactagaga acccactgct tactggctta tcgaaattaa
     2941 tacgactcac tatagggaga cccaagcttg gtcgaccaat tccgcagcag cagctcccgc
     3001 ggctcctgct ctgctccgcc tcggccccgg agcgaggggc ggagagccgc gcgcttgcct
     3061 tagtccgagc cgccaccctc cccgcgttcc ccgctttccc ggcccgcccg aggccgcccg
     3121 ccccgtcccc gccgccccgc agcccggccg cgccccgccg ccgccatggg ctgcctcggc
     3181 aacagtaaga ccgaggacca gcgcaacgag gagaaggcgc agcgcgaggc caacaaaaag
     3241 atcgagaagc agctgcagaa ggacaagcag gtctaccggg ccacgcaccg cctgctgctg
     3301 ctgggtgctg gagagtctgg caaaagcacc attgtgaagc agatgaggat tctacatgtt
     3361 aatgggttta acggagaggg cggcgaagag gacccgcagg ctgcaaggag caacagcgat
     3421 ggtgagaagg ccaccaaagt gcaggacatc aaaaacaacc tgaaggaggc cattgaaacc
     3481 attgtggccg ccatgagcaa cctggtgccc cccgtggagc tggccaaccc tgagaaccag
     3541 ttcagagtgg actacattct gagcgtgatg aacgtgccaa actttgactt cccacctgaa
     3601 ttctatgagc atgccaaggc tctgtgggag gatgagggag ttcgtgcctg ctacgagcgc
     3661 tccaacgagt accagctgat cgactgtgcc cagtacttcc tggacaagat tgatgtgatc
     3721 aagcaggccg agtacatgcc gactgagcag gacctgcttc gctgccgcgt cctgacctct
     3781 ggaatctttg agaccaagtt ccaggtggac aaagtcaact tccacatgtt cgatgtgggc
     3841 gcccagcgcg atgaacgccg caagtggatt cagtgcttca atgatgtgac tgccatcatc
     3901 ttcgtggtgg ccagcagcag ctacaacatg gtcatccggg aggacaacca gaccaaccgt
     3961 ctgcaggagg ctctgaagct gttcgacagc atctggaaca acaaatggct gcgtgacaca
     4021 tctgtgatcc tcttcctcaa caagcaagat ctgcttgctg agaaggtcct cgctgggaaa
     4081 tcgaagattg aggactactt tccagagttc gctcgctaca ccactcctga ggatgcgact
     4141 cccgagcccg gagaggaccc acgcgtgacc cgggccaagt acttcatccg ggatgagttt
     4201 ctgagaatca gcactgctag tggagatgga cgtcactact gctaccctca ctttacctgc
     4261 gccgtggaca ctgagaacat ccgccgtgtc ttcaacgact gccgtgacat catccagcgc
     4321 atgcatcttc gccaatacga gctgctctaa gaagggaacg cccaaattta attcagcctt
     4381 aagcacaatt aattaagagt gaaacgcaat cgtacaagca gttgatcacc caccataggg
     4441 catgatcaac accgcaacct ttcccttttc tccccagtga ttctgaaaac cccctcttcc
     4501 cttcagcttg cttagatgtt ccaaatttag taaagctcca agcttggcca agcttggtac
     4561 cgagctcgga tccactagta acggccgcca gtgtgctgga attctgcaga tatccatcac
     4621 actggcggcc gctcgagcat gcatctagag ggccctattc tatagtgtca cctaaatgct
     4681 agaggatctt tgtgaaggaa ccttacttct gtggtgtgac ataattggac aaactaccta
     4741 cagagattta aagctctaag gtaaatataa aatttttaag tgtataatgt gttaaactac
     4801 tgattctaat tgtttgtgta ttttagattc caacctatgg aactgatgaa tgggagcagt
     4861 ggtggaatgc ctttaatgag gaaaacctgt tttgctcaga agaaatgcca tctagtgatg
     4921 atgaggctac tgctgactct caacattcta ctcctccaaa aaagaagaga aaggtagaag
     4981 accccaagga ctttccttca gaattgctaa gttttttgag tcatgctgtg tttagtaata
     5041 gaactcttgc ttgctttgct atttacacca caaaggaaaa agctgcactg ctatacaaga
     5101 aaattatgga aaaatatttg atgtatagtg ccttgactag agatcataat cagccatacc
     5161 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     5221 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     5281 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     5341 ggtttgtcca aactcatcaa tgtatcttat catgtctgga tcatcccgcc atggtatcaa
     5401 cgccatattt ctatttacag tagggacctc ttcgttgtgt aggtaccgct gtattcctag
     5461 ggaaatagta gaggcacctt gaactgtctg catcagccat atagcccccg ctgttcgact
     5521 tacaaacaca ggcacagtac tgacaaaccc atacacctcc tctgaaatac ccatagttgc
     5581 tagggctgtc tccgaactca ttacaccctc caaagtcaga gctgtaattt cgccatcaag
     5641 ggcagcgagg gcttctccag ataaaatagc ttctgccgag agtcccgtaa gggtagacac
     5701 ttcagctaat ccctcgatga ggtctactag aatagtcagt gcggctccca ttttgaaaat
     5761 tcacttactt gatcagcttc agaagatggc ggagggcctc caacacagta attttcctcc
     5821 cgactcttaa aatagaaaat gtcaagtcag ttaagcagga agtggactaa ctgacgcagc
     5881 tggccgtgcg acatcctctt ttaattagtt gctaggcaac gccctccaga gggcgtgtgg
     5941 ttttgcaaga ggaagcaaaa gcctctccac ccaggcctag aatgtttcca cccaatcatt
     6001 actatgacaa cagctgtttt ttttagtatt aagcagaggc cggggacccc tgggcccgct
     6061 tactctggag aaaaagaaga gaggcattgt agaggcttcc agaggcaact tgtcaaaaca
     6121 ggactgcttc tatttctgtc acactgtctg gccctgtcac aaggtccagc acctccatac
     6181 cccctttaat aagcagtttg ggaacgggtg cgggtcttac tccgcccatc ccgcccctaa
     6241 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     6301 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     6361 gcctaggctt ttgcaaaaag ctaattc
//
