LOCUS       sequence-119279-        7107 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This G protein alpha-s construct contains internal insertions of YFP
            and the EE epitope..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7107)
  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 7107)
  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-s-YFP
FEATURES             Location/Qualifiers
     source          1..7107
                     /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             join(3400..3402,3403..3405,3406..4116)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3445..3466)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3706..3725)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4053..4074
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(5379..5397)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5380..5397)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     intron          5481..5546
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             5676..5696
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    5968..6102
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6005..6024)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6059..6078
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6948..7083
                     /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 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     3481 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     3541 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cttcggctac
     3601 ggcctgatgt gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     3661 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     3721 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat tgagctgaag
     3781 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     3841 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     3901 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     3961 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccgcc
     4021 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     4081 gccgggatca ctctcggcat ggacgagctg tacaagtccg gaggtggagg atccgatggt
     4141 gagaaggcca ccaaagtgca ggacatcaaa aacaacctga aggaggccat tgaaaccatt
     4201 gtggccgcca tgagcaacct ggtgcccccc gtggagctgg ccaaccctga gaaccagttc
     4261 agagtggact acattctgag cgtgatgaac gtgccaaact ttgacttccc acctgaattc
     4321 tatgagcatg ccaaggctct gtgggaggat gagggagttc gtgcctgcta cgagcgctcc
     4381 aacgagtacc agctgatcga ctgtgcccag tacttcctgg acaagattga tgtgatcaag
     4441 caggccgagt acatgccgac tgagcaggac ctgcttcgct gccgcgtcct gacctctgga
     4501 atctttgaga ccaagttcca ggtggacaaa gtcaacttcc acatgttcga tgtgggcggc
     4561 cagcgcgatg aacgccgcaa gtggattcag tgcttcaatg atgtgactgc catcatcttc
     4621 gtggtggcca gcagcagcta caacatggtc atccgggagg acaaccagac caaccgtctg
     4681 caggaggctc tgaacctctt caagagcatc tggaacaaca gatggctgcg taccatctct
     4741 gtgatcctct tcctcaacaa gcaagatctg cttgctgaga aggtcctcgc tgggaaatcg
     4801 aagattgagg actactttcc agagttcgct cgctacacca ctcctgagga tgcgactccc
     4861 gagcccggag aggacccacg cgtgacccgg gccaagtact tcatccggga tgagtttctg
     4921 agaatcagca ctgctagtgg agatggacgt cactactgct accctcactt tacctgcgcc
     4981 gtggacactg agaacatccg ccgtgtcttc aacgactgcc gtgacatcat ccagcgcatg
     5041 catcttcgcc aatacgagct gctctaagaa gggaacgccc aaatttaatt cagccttaag
     5101 cacaattaat taagagtgaa acgcaatcgt acaagcagtt gatcacccac catagggcat
     5161 gatcaacacc gcaacctttc ccttttctcc ccagtgattc tgaaaacccc ctcttccctt
     5221 cagcttgctt agatgttcca aatttagtaa gctccaagct tggccaagct tggtaccgag
     5281 ctcggatcga tccactagta acggccgcca gtgtgctgga attctgcaga tatccatcac
     5341 actggcggcc gctcgagcat gcatctagag ggccctattc tatagtgtca cctaaatgct
     5401 agaggatctt tgtgaaggaa ccttacttct gtggtgtgac ataattggac aaactaccta
     5461 cagagattta aagctctaag gtaaatataa aatttttaag tgtataatgt gttaaactac
     5521 tgattctaat tgtttgtgta ttttagattc caacctatgg aactgatgaa tgggagcagt
     5581 ggtggaatgc ctttaatgag gaaaacctgt tttgctcaga agaaatgcca tctagtgatg
     5641 atgaggctac tgctgactct caacattcta ctcctccaaa aaagaagaga aaggtagaag
     5701 accccaagga ctttccttca gaattgctaa gttttttgag tcatgctgtg tttagtaata
     5761 gaactcttgc ttgctttgct atttacacca caaaggaaaa agctgcactg ctatacaaga
     5821 aaattatgga aaaatatttg atgtatagtg ccttgactag agatcataat cagccatacc
     5881 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     5941 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     6001 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     6061 ggtttgtcca aactcatcaa tgtatcttat catgtctgga tcatcccgcc atggtatcaa
     6121 cgccatattt ctatttacag tagggacctc ttcgttgtgt aggtaccgct gtattcctag
     6181 ggaaatagta gaggcacctt gaactgtctg catcagccat atagcccccg ctgttcgact
     6241 tacaaacaca ggcacagtac tgacaaaccc atacacctcc tctgaaatac ccatagttgc
     6301 tagggctgtc tccgaactca ttacaccctc caaagtcaga gctgtaattt cgccatcaag
     6361 ggcagcgagg gcttctccag ataaaatagc ttctgccgag agtcccgtaa gggtagacac
     6421 ttcagctaat ccctcgatga ggtctactag aatagtcagt gcggctccca ttttgaaaat
     6481 tcacttactt gatcagcttc agaagatggc ggagggcctc caacacagta attttcctcc
     6541 cgactcttaa aatagaaaat gtcaagtcag ttaagcagga agtggactaa ctgacgcagc
     6601 tggccgtgcg acatcctctt ttaattagtt gctaggcaac gccctccaga gggcgtgtgg
     6661 ttttgcaaga ggaagcaaaa gcctctccac ccaggcctag aatgtttcca cccaatcatt
     6721 actatgacaa cagctgtttt ttttagtatt aagcagaggc cggggacccc tgggcccgct
     6781 tactctggag aaaaagaaga gaggcattgt agaggcttcc agaggcaact tgtcaaaaca
     6841 ggactgcttc tatttctgtc acactgtctg gccctgtcac aaggtccagc acctccatac
     6901 cccctttaat aagcagtttg ggaacgggtg cgggtcttac tccgcccatc ccgcccctaa
     6961 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     7021 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     7081 gcctaggctt ttgcaaaaag ctaattc
//
