LOCUS       sequence-336178-        7254 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding G-alpha-13 internally-tagged with the Citrine 
            variant of YFP in the b/c loop.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7254)
  AUTHORS   Maziarz M, Park JC, Leyme A, Marivin A, Garcia-Lopez A, Patel PP, 
            Garcia-Marcos M
  TITLE     Revealing the Activity of Trimeric G-proteins in Live Cells with a 
            Versatile Biosensor Design.
  JOURNAL   Cell. 2020 Jun 27. pii: S0092-8674(20)30752-2. doi: 
            10.1016/j.cell.2020.06.020.
  PUBMED    32634377
REFERENCE   2  (bases 1 to 7254)
  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: "Cell. 2020 
            Jun 27. pii: S0092-8674(20)30752-2. doi: 
            10.1016/j.cell.2020.06.020."
COMMENT     Sequence Label: pcDNA3.1-G-alpha-13(EE)-YFP
FEATURES             Location/Qualifiers
     source          1..7254
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(189..208)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     536..557
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1400..1417)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1406..1630
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1676..2104
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1763..1782)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1973..1994
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2113..2133)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2118..2447
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2298..2433
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2360..2379
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2514..3308
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2568..2587)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3178..3197
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3482..3603
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3519..3538)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3573..3592
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3652..3668)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3652..3668)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3665..3687)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3676..3692
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3700..3706,3707..3724,3725..3730))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3745..3766
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3883..3900)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4054..4642)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4134..4153)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4813..5604,5605..5673))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGVGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5436..5455
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5674..5778)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5853..5872)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6044..6423
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6424..6627
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6578..6598
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6672..6691
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6672..6690
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 atggaaaact gacccttaaa tttatttgta ctacgggtaa attgcccgtt ccctggccga
       61 cattggtaac tacattcgga tatggtttga tgtgctttgc aaggtatccg gatcacatga
      121 agcagcatga tttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
      181 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
      241 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
      301 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
      361 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
      421 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
      481 actacctgag ctaccagtcc gccatgagca aagaccccaa cgagaagcgc gatcacatgg
      541 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagg
      601 gatcaggccc gcggatggca gcccaaggaa tggtggaaac aagggttttc ttacaatatc
      661 ttcctgctat aagagcatta tgggcagaca gcggcataca gaatgcctat gaccggcgtc
      721 gagaatttca actgggtgaa tctgtaaaat atttcctgga taacttggat aaacttggag
      781 aaccagagta tatgccaaca gaacaagata ttctgcttgc cagaagaccc accaaaggca
      841 tccatgaata cgactttgaa ataaaaaatg ttcctttcaa aatggttgat gtaggtggtc
      901 agagatcaga aaggaaacgt tggtttgaat gtttcgacag tgtgacatca atacttttcc
      961 ttgtttcctc aagtgaattt gaccaggtgc ttatggaaga tcgactgacc aatcgcctta
     1021 cagagtctct gaacattttt gaaacaatcg tcaataaccg ggttttcagc aatgtctcca
     1081 taattctgtt cttaaacaag acagacttgc ttgaggagaa ggtgcaaatt gtgagcatca
     1141 aagactattt cctagaattt gaaggggatc cccactgctt aagagacgtc caaaaattcc
     1201 tggtggaatg tttccggaac aaacgccggg accagcaaca gaagccctta taccaccact
     1261 tcaccactgc tatcaacacg gagaacatcc gccttgtttt ccgtgacgtg aaggatacta
     1321 ttctgcatga caacctcaag cagcttatgc tacagtgatc tagagatctc tagagggccc
     1381 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     1441 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     1501 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     1561 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     1621 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     1681 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     1741 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     1801 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     1861 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     1921 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     1981 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     2041 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     2101 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     2161 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     2221 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     2281 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     2341 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     2401 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     2461 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     2521 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     2581 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     2641 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     2701 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     2761 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     2821 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     2881 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     2941 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     3001 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     3061 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     3121 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     3181 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     3241 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     3301 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     3361 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     3421 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     3481 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     3541 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     3601 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     3661 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     3721 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     3781 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     3841 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     3901 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     3961 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4021 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4081 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4141 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4201 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4261 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4321 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4381 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4441 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     4501 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4561 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4621 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4681 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     4741 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     4801 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4861 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     4921 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccga ctccagattt
     4981 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5041 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5101 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5161 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5221 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5281 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5341 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5401 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5461 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     5521 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5581 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     5641 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     5701 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     5761 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     5821 agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     5881 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
     5941 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg
     6001 cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact
     6061 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     6121 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     6181 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     6241 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     6301 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     6361 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     6421 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
     6481 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
     6541 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
     6601 cggtgggagg tctatataag cagagctctc tggctaacta gagaacccac tgcttactgg
     6661 cttatcgaaa ttaatacgac tcactatagg gagacccaag ctggctagcg tttaaactta
     6721 agcttggtac caccatggcg gacttcctgc cgtcgcggtc cgtgctgtcc gtgtgcttcc
     6781 ccggctgcct gctgacgagt ggcgaggccg agcagcaacg caagtccaag gagatcgaca
     6841 aatgcctgtc tcgggaaaag acctatgtga agcggctggt gaagatcctg ctgctgggcg
     6901 cgggcgagag cggcaagtcc accttcctga agcagatgcg gatcatccac gggcaggact
     6961 tcgaccagcg cgcgcgcgag gagttccgcc ccaccatcta cagcaacgtg atcaaaggta
     7021 tgagggtgct ggttgatgct cgagagaagc ttcatattcc ctggggagac aactcaaacc
     7081 aacaacatgg agataagatg atgtcgtttg atacccgggc cccgcgggga tcaggcatgg
     7141 tttcaaaggg agaagagttg ttcacagggg ttgtaccgat tcttgtagaa cttgacggag
     7201 atgtcaacgg gcacaagttt agtgtaagtg gagaaggcga gggcgatgcg acct
//
