LOCUS       sequence-123362-        7170 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The heterotrimeric G-protein subunit, G-alpha-i isoform 2, tagged 
            with mTurquoise2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7170)
  AUTHORS   van Unen J, Stumpf AD, Schmid B, Reinhard NR, Hordijk PL, Hoffmann 
            C, Gadella TW Jr, Goedhart J
  TITLE     A New Generation of FRET Sensors for Robust Measurement of Galphai1,
            Galphai2 and Galphai3 Activation Kinetics in Single Cells.
  JOURNAL   PLoS One. 2016 Jan 22;11(1):e0146789. doi: 
            10.1371/journal.pone.0146789. eCollection 2016.
  PUBMED    26799488
REFERENCE   2  (bases 1 to 7170)
  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: "PLoS One.";
            date: "2016-01-22"; volume: "11(1):e0146789. doi"; pages: " 
            10.1371/journal.pone.0146789. eCollection 2016"
COMMENT     Sequence Label: Galphai2-mTurquoise2
FEATURES             Location/Qualifiers
     source          1..7170
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(1314..1316,1317..1319,1320..1832)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIE"
     primer_bind     complement(1359..1380)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1620..1639)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1967..1988
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2764..2781)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2770..2994
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3040..3468
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3127..3146)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3337..3358
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3477..3497)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3482..3811
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3662..3797
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3724..3743
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3878..4672
                     /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(3932..3951)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4542..4561
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4846..4967
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4883..4902)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4937..4956
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5016..5032)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5016..5032)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5029..5051)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5040..5056
                     /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(5064..5070,5071..5088,5089..5094))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5109..5130
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5247..5264)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5418..6003)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5498..5517)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6174..6965,6966..7034))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6797..6816
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7035..7139)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaacgg gccctctaga ctcgagcggc cgccactgtg ctggatatct gcagaattca
      961 ccatgggctg caccgtgagc gccgaggaca aggcggcggc cgagcgctct aagatgatcg
     1021 acaagaacct gcgggaggac ggagagaagg cggcgcggga ggtgaagttg ctgctgttgg
     1081 gtgctgggga gtcagggaag agcaccatcg tcaagcagat gaagatcatc cacgaggatg
     1141 gctactccga ggaggaatgc cggcagtacc gggcggttgt ctacagcaac accatccagt
     1201 ccatcatggc cattgtcaaa gccatgggca acctgcagat cgactttgcc gacccctcca
     1261 gagcggacga cgccaggcag ctatttgcac tgtcctgcac cgccaccggt tctatggtga
     1321 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
     1381 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
     1441 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     1501 ccaccctgtc ctggggcgtg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg
     1561 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     1621 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     1681 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     1741 agtacaacta ctttagcgac aacgtctata tcaccgccga caagcagaag aacggcatca
     1801 aggccaactt caagatccgc cacaacatcg aggacggcgg cgtgcagctc gccgaccact
     1861 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     1921 gcacccagtc caagctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     1981 agttcgtgac cgccgccggg atcaccggtg aggagcaagg cgtgctccct gatgacctgt
     2041 ccggcgtcat ccggaggctc tgggctgacc atggtgtgca ggcctgcttt ggccgctcaa
     2101 gggaatacca gctcaacgac tcagctgcct actacctgaa cgacctggag cgtattgcac
     2161 agagtgacta catccccaca cagcaagatg tgctacggac ccgcgtaaag accacgggga
     2221 tcgtggagac acacttcacc ttcaaggacc tacacttcaa gatgtttgat gtgggtggtc
     2281 agcggtctga gcggaagaag tggatccact gctttgaggg cgtcacagcc atcatcttct
     2341 gcgtagcctt gagcgcctat gacttggtgc tagctgagga cgaggagatg aaccgcatgc
     2401 atgagagcat gaagctattc gatagcatct gcaacaacaa gtggttcaca gacacgtcca
     2461 tcatcctctt cctcaacaag aaggacctgt ttgaggagaa gatcacacac agtcccctga
     2521 ccatctgctt ccctgagtac acaggggcca acaaatatga tgaggcagcc agctacatcc
     2581 agagtaagtt tgaggacctg aataagcgca aagacaccaa ggagatctac acgcacttca
     2641 cgtgcgccac cgacaccaag aacgtgcagt tcgtgtttga cgccgtcacc gatgtcatca
     2701 tcaagaacaa cctgaaggac tgcggcctct tctgaagctt aataagttta aaccgctgat
     2761 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     2821 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     2881 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     2941 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     3001 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     3061 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     3121 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3181 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     3241 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     3301 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     3361 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     3421 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     3481 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3541 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     3601 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     3661 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     3721 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     3781 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     3841 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     3901 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     3961 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     4021 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     4081 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     4141 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     4201 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     4261 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     4321 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     4381 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     4441 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     4501 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     4561 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     4621 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     4681 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     4741 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     4801 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     4861 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     4921 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     4981 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     5041 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     5101 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     5161 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     5221 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     5281 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     5341 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5401 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5461 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5521 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5581 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5641 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5701 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5761 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5821 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     5881 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5941 accgctggta gcggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     6001 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     6061 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     6121 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     6181 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     6241 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     6301 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     6361 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     6421 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     6481 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     6541 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     6601 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     6661 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     6721 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     6781 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     6841 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     6901 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     6961 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     7021 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     7081 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     7141 acatttcccc gaaaagtgcc acctgacgtc
//
