LOCUS       sequence-112541-        7639 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This G protein alpha-o 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 7639)
  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 7639)
  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-YFP
FEATURES             Location/Qualifiers
     source          1..7639
                     /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      3726..3735
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4110..4112,4113..4115,4116..4826)
                     /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(4155..4176)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4416..4435)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4763..4784
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(5911..5929)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5912..5929)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     intron          6013..6078
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             6208..6228
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    6500..6634
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6537..6556)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6591..6610
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7480..7615
                     /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 atcgatctgc acccctcacc
     3121 cccggcccgg ccctctgccc agccctgccc tgcgcgcggg ggtcggagaa ggcgccgcgg
     3181 acgcaccgac ggccgaggag cggcgatgca catgcactag cggcaccccc taactcactc
     3241 cctccacacc ccgcgccgcc gccgccgcca ccgcctccgc ctccgcctcc tcctccgcct
     3301 ccggcagccg cggcagaagg acccaccctg ccccccaccc caccctccgc cggctccggc
     3361 tgcggatcca gcctcgactc ctattttatt tattttgggt cgtgcactag tctcggtgcc
     3421 tgcaacccgc gcctcccggg cccgcgggcg cctcctctct cggctccgga gccccagacc
     3481 ccggccaccc tcacctcgac acccccagac cccagccagc cgccgctaat cttcgccgct
     3541 ggaatcttga tagaggctgt ccttttgggg ggattctggt ctttcgacaa ttttgttccc
     3601 aaccaaggaa aggatatcgt gattttctcc cctttgagcc caggctctgc tctgtggggg
     3661 ggtggggggc gcgccgaccc gaggagtcgt gccagccgag tcgtgcgggc tgtggcaggg
     3721 aaggggccac catgggatgt actctgagcg cagaggagag agccgccctc gagcggagca
     3781 aggcgattga gaaaaatctc aaagaagatg gcatcagcgc cgccaaagac gtgaaattac
     3841 tcctgctggg ggctggagaa tcaggaaaaa gcaccattgt gaagcagatg aagatcatcc
     3901 atgaagatgg cttctctgga gaagacgtaa agcagtacaa gcctgtcgtc tacagcaaca
     3961 ccatccagtc tctggcagcc attgtgcggg ccatggatac tctgggcgtg gagtatggtg
     4021 acaaggagag gaaggcagac tccaagatgg tgtgtgacgt ggtgagtcgc atggaggaca
     4081 ctgaaccaag atccggtggg ggcgggagca tggtgagcaa gggcgaggag ctgttcaccg
     4141 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     4201 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     4261 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cttcggctac ggcctgatgt
     4321 gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     4381 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     4441 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat tgagctgaag ggcatcgact
     4501 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     4561 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     4621 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     4681 acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccgcc ctgagcaaag
     4741 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     4801 ctctcggcat ggacgagctg tacaagtccg gaggtggagg atctttctct gcagaactgc
     4861 tttctgccat gatgcgactc tggggcgact cggggatcca ggagtgcttc aaccgatctc
     4921 gggagtatca gctcaacgac tctgccaaat actacctgga cagcttggat cggattggag
     4981 ctgcagaata catgcccacc gagcaggaca tcctccgaac cagggtcaaa acaactggca
     5041 tcgtagaaac ccacttcacc ttcaagaacc tccacttcag gctgtttgac gttgggggcc
     5101 agcgatctga acgtaagaag tggatccact gcttcgagga tgtcacggcc atcatcttct
     5161 gtgtcgcact cagcggctat gaccaggtgc tccacgagga cgaaaccacg aaccgcatgc
     5221 acgagtctct catgctcttc gactccatct gtaacaacaa gtttttcatc gatacctcca
     5281 tcattctctt cctcaacaag aaagacctct ttggcgagaa gattaagaag tcacccttga
     5341 ccatctgctt tcctgaatac ccaggctcca acacctatga agacgcagct gcctacatcc
     5401 aaacacagtt tgaaagcaaa aaccgctcac ccaacaaaga aatttactgt cacatgactt
     5461 gtgccacaga cacgaataat atccaggtgg tattcgacgc cgtcaccgac atcatcattg
     5521 ccaacaatct ccggggctgt ggcttgtact gacctcttgt cctgtatagc aacctatttg
     5581 actgcttcat ggactctttg ctgttgatgt tgatctcctg gtagcatgac ctttggcctt
     5641 tgtaagacac acagcctttc tgtaccaagc ccctgtctaa cctacgaccc cagagtgact
     5701 gacggctgtg tatttctgta gaatgctgta gaatacggtt ttagttgagt ctttacattt
     5761 agaacttgaa aggatttaaa aaaaaaaaaa catttctcat gtgctttgta gctttaaaaa
     5821 ggaaaactca ccatttcatc catatttccg gaattctgca gatatccatc acactggcgg
     5881 ccgctcgagc atgcatctag agggccctat tctatagtgt cacctaaatg ctagaggatc
     5941 tttgtgaagg aaccttactt ctgtggtgtg acataattgg acaaactacc tacagagatt
     6001 taaagctcta aggtaaatat aaaattttta agtgtataat gtgttaaact actgattcta
     6061 attgtttgtg tattttagat tccaacctat ggaactgatg aatgggagca gtggtggaat
     6121 gcctttaatg aggaaaacct gttttgctca gaagaaatgc catctagtga tgatgaggct
     6181 actgctgact ctcaacattc tactcctcca aaaaagaaga gaaaggtaga agaccccaag
     6241 gactttcctt cagaattgct aagttttttg agtcatgctg tgtttagtaa tagaactctt
     6301 gcttgctttg ctatttacac cacaaaggaa aaagctgcac tgctatacaa gaaaattatg
     6361 gaaaaatatt tgatgtatag tgccttgact agagatcata atcagccata ccacatttgt
     6421 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     6481 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     6541 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     6601 caaactcatc aatgtatctt atcatgtctg gatcatcccg ccatggtatc aacgccatat
     6661 ttctatttac agtagggacc tcttcgttgt gtaggtaccg ctgtattcct agggaaatag
     6721 tagaggcacc ttgaactgtc tgcatcagcc atatagcccc cgctgttcga cttacaaaca
     6781 caggcacagt actgacaaac ccatacacct cctctgaaat acccatagtt gctagggctg
     6841 tctccgaact cattacaccc tccaaagtca gagctgtaat ttcgccatca agggcagcga
     6901 gggcttctcc agataaaata gcttctgccg agagtcccgt aagggtagac acttcagcta
     6961 atccctcgat gaggtctact agaatagtca gtgcggctcc cattttgaaa attcacttac
     7021 ttgatcagct tcagaagatg gcggagggcc tccaacacag taattttcct cccgactctt
     7081 aaaatagaaa atgtcaagtc agttaagcag gaagtggact aactgacgca gctggccgtg
     7141 cgacatcctc ttttaattag ttgctaggca acgccctcca gagggcgtgt ggttttgcaa
     7201 gaggaagcaa aagcctctcc acccaggcct agaatgtttc cacccaatca ttactatgac
     7261 aacagctgtt ttttttagta ttaagcagag gccggggacc cctgggcccg cttactctgg
     7321 agaaaaagaa gagaggcatt gtagaggctt ccagaggcaa cttgtcaaaa caggactgct
     7381 tctatttctg tcacactgtc tggccctgtc acaaggtcca gcacctccat acccccttta
     7441 ataagcagtt tgggaacggg tgcgggtctt actccgccca tcccgcccct aactccgccc
     7501 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     7561 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     7621 ttttgcaaaa agctaattc
//
