LOCUS       sequence-122021-        6885 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This is a constitutively activated G protein alpha-o mutant 
            containing an internal EE epitope..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6885)
  AUTHORS   Yost EA, Mervine SM, Sabo JL, Hynes TR, Berlot CH
  TITLE     Live cell analysis of G protein beta5 complex formation, function, 
            and targeting.
  JOURNAL   Mol Pharmacol. 2007 Oct;72(4):812-25. Epub 2007 Jun 27.
  PUBMED    17596375
REFERENCE   2  (bases 1 to 6885)
  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: "Mol 
            Pharmacol."; date: "2007-10"; volume: "72(4)"; pages: "812-25. Epub 
            2007 Jun 27"
COMMENT     Sequence Label: G protein alpha-o-R179C-EE
FEATURES             Location/Qualifiers
     source          1..6885
                     /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      3722..3731
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5157..5175)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5158..5175)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     intron          5259..5324
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             5454..5474
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    5746..5880
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5783..5802)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5837..5856
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6726..6861
                     /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 gtaccgagct cggatccact agtaacggcc
     3001 gccagtgtgc tggaattccc tctcgcgctc tccctgtctc ctgtccgctc cgccgagcga
     3061 tgcgagttct tggccccggc gacgccgcct ccagctagag atctgcaccc ctcacccccg
     3121 gcccggccct ctgcccagcc ctgccctgcg cgcgggggtc ggagaaggcg ccgcggacgc
     3181 accgacggcc gaggagcggc gatgcacatg cactagcggc accccctaac tcactccctc
     3241 cacaccccgc gccgccgccg ccgccaccgc ctccgcctcc gcctcctcct ccgcctccgg
     3301 cagccgcggc agaaggaccc accctgcccc ccaccccacc ctccgccggc tccggctgcg
     3361 gatccagcct cgactcctat tttatttatt ttgggtcgtg cactagtctc ggtgcctgca
     3421 acccgcgcct cccgggcccg cgggcgcctc ctctctcggc tccggagccc cagaccccgg
     3481 ccaccctcac ctcgacaccc ccagacccca gccagccgcc gctaatcttc gccgctggaa
     3541 tcttgataga ggctgtcctt ttggggggat tctggtcttt cgacaatttt gttcccaacc
     3601 aaggaaagga tatcgtgatt ttctcccctt tgagcccagg ctctgctctg tgggggggtg
     3661 gggggcgcgc cgacccgagg agtcgtgcca gccgagtcgt gcgggctgtg gcagggaagg
     3721 ggccaccatg ggatgtactc tgagcgcaga ggagagagcc gccctcgagc ggagcaaggc
     3781 gattgagaaa aatctcaaag aagatggcat cagcgccgcc aaagacgtga aattactcct
     3841 gctgggggct ggagaatcag gaaaaagcac cattgtgaag cagatgaaga tcatccatga
     3901 agatggcttc tctggagaag acgtaaagca gtacaagcct gtcgtctaca gcaacaccat
     3961 ccagtctctg gcagccattg tgcgggccat ggatactctg ggcgtggagt atggtgacaa
     4021 ggagaggaag gcagactcca agatggtgtg tgacgtggtg agtcgcatgg aggacactga
     4081 accaagatct ttctctgcag aactgctttc tgccatgatg cgactctggg gcgactcggg
     4141 gatccaggag tgcttcaacc gatctcggga gtatcagctc aacgactctg ccaaatacta
     4201 cctggacagc ttggatcgga ttggagccgc tgaatacatg cccaccgagc aggatatcct
     4261 ccgaacctgc gtcaaaacaa ctggcatcgt agaaacccac ttcaccttca agaacctcca
     4321 cttcaggctg tttgacgttg ggggccagcg atctgaacgt aagaagtgga tccactgctt
     4381 cgaggatgtc acggccatca tcttctgtgt cgcactcagc ggctatgacc aggtgctcca
     4441 cgaggacgaa accacgaacc gcatgcacga gtctctcatg ctcttcgact ccatctgtaa
     4501 caacaagttt ttcatcgata cctccatcat tctcttcctc aacaagaaag acctctttgg
     4561 cgagaagatt aagaagtcac ccttgaccat ctgctttcct gaatacccag gctccaacac
     4621 ctatgaagac gcagctgcct acatccaaac acagtttgaa agcaaaaacc gctcacccaa
     4681 caaagaaatt tactgtcaca tgacttgtgc cacagacacg aataatatcc aggtggtatt
     4741 cgacgccgtc accgacatca tcattgccaa caatctccgg ggctgtggct tgtactgacc
     4801 tcttgtcctg tatagcaacc tatttgactg cttcatggac tctttgctgt tgatgttgat
     4861 ctcctggtag catgaccttt ggcctttgta agacacacag cctttctgta ccaagcccct
     4921 gtctaaccta cgaccccaga gtgactgacg gctgtgtatt tctgtagaat gctgtagaat
     4981 acggttttag ttgagtcttt acatttagaa cttgaaagga tttaaaaaaa aaaaaacatt
     5041 tctcatgtgc tttgtagctt taaaaaggaa aactcaccat ttcatccata tttccggaat
     5101 tctgcagata tccatcacac tggcggccgc tcgagcatgc atctagaggg ccctattcta
     5161 tagtgtcacc taaatgctag aggatctttg tgaaggaacc ttacttctgt ggtgtgacat
     5221 aattggacaa actacctaca gagatttaaa gctctaaggt aaatataaaa tttttaagtg
     5281 tataatgtgt taaactactg attctaattg tttgtgtatt ttagattcca acctatggaa
     5341 ctgatgaatg ggagcagtgg tggaatgcct ttaatgagga aaacctgttt tgctcagaag
     5401 aaatgccatc tagtgatgat gaggctactg ctgactctca acattctact cctccaaaaa
     5461 agaagagaaa ggtagaagac cccaaggact ttccttcaga attgctaagt tttttgagtc
     5521 atgctgtgtt tagtaataga actcttgctt gctttgctat ttacaccaca aaggaaaaag
     5581 ctgcactgct atacaagaaa attatggaaa aatatttgat gtatagtgcc ttgactagag
     5641 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     5701 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     5761 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     5821 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc
     5881 atcccgccat ggtatcaacg ccatatttct atttacagta gggacctctt cgttgtgtag
     5941 gtaccgctgt attcctaggg aaatagtaga ggcaccttga actgtctgca tcagccatat
     6001 agcccccgct gttcgactta caaacacagg cacagtactg acaaacccat acacctcctc
     6061 tgaaataccc atagttgcta gggctgtctc cgaactcatt acaccctcca aagtcagagc
     6121 tgtaatttcg ccatcaaggg cagcgagggc ttctccagat aaaatagctt ctgccgagag
     6181 tcccgtaagg gtagacactt cagctaatcc ctcgatgagg tctactagaa tagtcagtgc
     6241 ggctcccatt ttgaaaattc acttacttga tcagcttcag aagatggcgg agggcctcca
     6301 acacagtaat tttcctcccg actcttaaaa tagaaaatgt caagtcagtt aagcaggaag
     6361 tggactaact gacgcagctg gccgtgcgac atcctctttt aattagttgc taggcaacgc
     6421 cctccagagg gcgtgtggtt ttgcaagagg aagcaaaagc ctctccaccc aggcctagaa
     6481 tgtttccacc caatcattac tatgacaaca gctgtttttt ttagtattaa gcagaggccg
     6541 gggacccctg ggcccgctta ctctggagaa aaagaagaga ggcattgtag aggcttccag
     6601 aggcaacttg tcaaaacagg actgcttcta tttctgtcac actgtctggc cctgtcacaa
     6661 ggtccagcac ctccataccc cctttaataa gcagtttggg aacgggtgcg ggtcttactc
     6721 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     6781 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     6841 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct aattc
//
