LOCUS       sequence-121428-        7098 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  G protein alpha-s internally tagged with mCherry and EE epitope.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7098)
  AUTHORS   Hynes TR, Mervine SM, Yost EA, Sabo JL, Berlot CH
  TITLE     Live cell imaging of Gs and the beta2-adrenergic receptor 
            demonstrates that both alphas and beta1gamma7 internalize upon 
            stimulation and exhibit similar trafficking patterns that differ 
            from that of the beta2-adrenergic receptor.
  JOURNAL   J Biol Chem. 2004 Oct 15;279(42):44101-12. Epub 2004 Aug 5.
  PUBMED    15297467
REFERENCE   2  (bases 1 to 7098)
  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: "2004-10-15"; volume: "279(42)"; pages: "44101-12. 
            Epub 2004 Aug 5"
COMMENT     Sequence Label: G protein alpha-s-mCherry
FEATURES             Location/Qualifiers
     source          1..7098
                     /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"
     CDS             3400..4107
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3545..3563)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3813..3832
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     promoter        complement(5370..5388)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5371..5388)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     intron          5472..5537
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             5667..5687
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    5959..6093
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5996..6015)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6050..6069
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6939..7074
                     /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 atccggtggg ggcgggagca tggtgagcaa gggcgaggag
     3421 gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg
     3481 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
     3541 accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc
     3601 cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac
     3661 ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc
     3721 ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg
     3781 aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc
     3841 tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag
     3901 cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag
     3961 gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc
     4021 tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc
     4081 accggcggca tggacgagct gtacaagtcc ggaggtggag gatccgatgg tgagaaggcc
     4141 accaaagtgc aggacatcaa aaacaacctg aaggaggcca ttgaaaccat tgtggccgcc
     4201 atgagcaacc tggtgccccc cgtggagctg gccaaccctg agaaccagtt cagagtggac
     4261 tacattctga gcgtgatgaa cgtgccaaac tttgacttcc cacctgaatt ctatgagcat
     4321 gccaaggctc tgtgggagga tgagggagtt cgtgcctgct acgagcgctc caacgagtac
     4381 cagctgatcg actgtgccca gtacttcctg gacaagattg atgtgatcaa gcaggccgag
     4441 tacatgccga ctgagcagga cctgcttcgc tgccgcgtcc tgacctctgg aatctttgag
     4501 accaagttcc aggtggacaa agtcaacttc cacatgttcg atgtgggcgg ccagcgcgat
     4561 gaacgccgca agtggattca gtgcttcaat gatgtgactg ccatcatctt cgtggtggcc
     4621 agcagcagct acaacatggt catccgggag gacaaccaga ccaaccgtct gcaggaggct
     4681 ctgaacctct tcaagagcat ctggaacaac agatggctgc gtaccatctc tgtgatcctc
     4741 ttcctcaaca agcaagatct gcttgctgag aaggtcctcg ctgggaaatc gaagattgag
     4801 gactactttc cagagttcgc tcgctacacc actcctgagg atgcgactcc cgagcccgga
     4861 gaggacccac gcgtgacccg ggccaagtac ttcatccggg atgagtttct gagaatcagc
     4921 actgctagtg gagatggacg tcactactgc taccctcact ttacctgcgc cgtggacact
     4981 gagaacatcc gccgtgtctt caacgactgc cgtgacatca tccagcgcat gcatcttcgc
     5041 caatacgagc tgctctaaga agggaacgcc caaatttaat tcagccttaa gcacaattaa
     5101 ttaagagtga aacgcaatcg tacaagcagt tgatcaccca ccatagggca tgatcaacac
     5161 cgcaaccttt cccttttctc cccagtgatt ctgaaaaccc cctcttccct tcagcttgct
     5221 tagatgttcc aaatttagta agctccaagc ttggccaagc ttggtaccga gctcggatcg
     5281 atccactagt aacggccgcc agtgtgctgg aattctgcag atatccatca cactggcggc
     5341 cgctcgagca tgcatctaga gggccctatt ctatagtgtc acctaaatgc tagaggatct
     5401 ttgtgaagga accttacttc tgtggtgtga cataattgga caaactacct acagagattt
     5461 aaagctctaa ggtaaatata aaatttttaa gtgtataatg tgttaaacta ctgattctaa
     5521 ttgtttgtgt attttagatt ccaacctatg gaactgatga atgggagcag tggtggaatg
     5581 cctttaatga ggaaaacctg ttttgctcag aagaaatgcc atctagtgat gatgaggcta
     5641 ctgctgactc tcaacattct actcctccaa aaaagaagag aaaggtagaa gaccccaagg
     5701 actttccttc agaattgcta agttttttga gtcatgctgt gtttagtaat agaactcttg
     5761 cttgctttgc tatttacacc acaaaggaaa aagctgcact gctatacaag aaaattatgg
     5821 aaaaatattt gatgtatagt gccttgacta gagatcataa tcagccatac cacatttgta
     5881 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     5941 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     6001 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     6061 aaactcatca atgtatctta tcatgtctgg atcatcccgc catggtatca acgccatatt
     6121 tctatttaca gtagggacct cttcgttgtg taggtaccgc tgtattccta gggaaatagt
     6181 agaggcacct tgaactgtct gcatcagcca tatagccccc gctgttcgac ttacaaacac
     6241 aggcacagta ctgacaaacc catacacctc ctctgaaata cccatagttg ctagggctgt
     6301 ctccgaactc attacaccct ccaaagtcag agctgtaatt tcgccatcaa gggcagcgag
     6361 ggcttctcca gataaaatag cttctgccga gagtcccgta agggtagaca cttcagctaa
     6421 tccctcgatg aggtctacta gaatagtcag tgcggctccc attttgaaaa ttcacttact
     6481 tgatcagctt cagaagatgg cggagggcct ccaacacagt aattttcctc ccgactctta
     6541 aaatagaaaa tgtcaagtca gttaagcagg aagtggacta actgacgcag ctggccgtgc
     6601 gacatcctct tttaattagt tgctaggcaa cgccctccag agggcgtgtg gttttgcaag
     6661 aggaagcaaa agcctctcca cccaggccta gaatgtttcc acccaatcat tactatgaca
     6721 acagctgttt tttttagtat taagcagagg ccggggaccc ctgggcccgc ttactctgga
     6781 gaaaaagaag agaggcattg tagaggcttc cagaggcaac ttgtcaaaac aggactgctt
     6841 ctatttctgt cacactgtct ggccctgtca caaggtccag cacctccata ccccctttaa
     6901 taagcagttt gggaacgggt gcgggtctta ctccgcccat cccgccccta actccgccca
     6961 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     7021 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     7081 tttgcaaaaa gctaattc
//
