LOCUS       sequence-266019-        6785 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Visualization of GRK2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6785)
  AUTHORS   Pietraszewska-Bogiel A, Joosen L, Chertkova AO, Goedhart J
  TITLE     Not So Dry After All: DRY Mutants of the AT1A Receptor and H1 
            Receptor Can Induce G-Protein-Dependent Signaling.
  JOURNAL   ACS Omega. 2020 Feb 3;5(6):2648-2659. doi: 10.1021/acsomega.9b03146.
            eCollection 2020 Feb 18.
  PUBMED    32095688
REFERENCE   2  (bases 1 to 6785)
  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"; doi: 
            "10.1021/acsomega.9b03146"; journalName: "ACS Omega"; date: 
            "2020-02-3- 3"; volume: "5"; issue: "6"; pages: "2648-2659"
COMMENT     Sequence Label: BtGRK2[D110A K220R]-mVenus
FEATURES             Location/Qualifiers
     source          1..6785
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2725..2734
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2731..2733,2734..2736,2737..3450)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2776..2797)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3037..3056)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3384..3405
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3573..3694
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3610..3629)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3664..3683
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3701..4156)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3838..3859)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4050..4069
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4183..4287
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4289..4646
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4289..4480
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4312..4332)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4497..4632
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4681..5475
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4681..5475
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4735..4754)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5345..5364
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5663..5682)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5707..5754
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6083..6671
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6572..6591
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc atggccgacc
      661 tggaggcggt gctggccgac gtgagctacc tgatggccat ggagaagagc aaggccacgc
      721 cggcggcgcg cgccagcaag aagatcctgc tgcccgagcc cagcatccgc agcgtcatgc
      781 agaagtacct ggaggaccgg ggcgaggtga cttttgagaa gatcttctcc cagaagctgg
      841 ggtacctgct tttccgagac ttctgcctga agcacctgga ggaggccaag cccttggtag
      901 agttctacga ggagatcaag aaatacgaga agctggagac agaggaggag cgcctggtct
      961 gcagccgaga gatcttcgcc acgtacatca tgaaggagct gctggcctgc tcacatcctt
     1021 tctcgaagag cgccattgag cacgtccagg gccatctggt gaagaagcag gtgcctccgg
     1081 atctcttcca gccatatatt gaagaaattt gccagaacct ccgaggagac gtgttccaga
     1141 aattcatcga gagcgataaa ttcacacgat tttgccagtg gaagaatgta gagctcaaca
     1201 tccacctgac catgaacgac ttcagtgtgc accgcatcat cgggcgaggc ggcttcggtg
     1261 aggtctacgg ctgccggaag gccgacacgg gcaagatgta cgccatgaag tgtctggaca
     1321 agaagcgcat caagatgaag caaggggaga ctctggccct gaatgagcgc atcatgctgt
     1381 cgctcgtcag caccggggac tgcccgttca tcgtctgcat gtcatacgcc ttccacacac
     1441 cggacaagct cagcttcatc ctggatctca tgaacggcgg ggacctgcac taccacctgt
     1501 cccagcacgg ggtcttctcc gaggccgaca tgcgtttcta cgccgccgag atcatcctgg
     1561 gcctggagca catgcacaac cgcttcgtgg tctaccggga cctgaagccg gccaacatcc
     1621 tgctggacga gcacggccac gtgcgcatct cagacctggg cctggcctgt gacttctcca
     1681 agaagaagcc tcacgccagt gtgggcaccc acgggtacat ggctcccgag gttctacaga
     1741 agggtgtggc ctacgacagc agcgccgact ggttctccct gggctgcatg ctcttcaagc
     1801 tgctgcgagg gcatagccct ttccggcagc acaagaccaa agacaagcat gagatcgaca
     1861 gaatgacatt gacaatggct gtggagctgc ctgactcctt ctcccctgag ctccgctcct
     1921 tgctggaggg gctgctgcag agggatgtca accggaggct aggctgcctg ggccgagggg
     1981 cccaggaggt gaaggagagc cccttcttcc gttccctgga ctggcagatg gtctttttac
     2041 aaaagtaccc tcccccgttg atccccccac gaggggaggt gaatgcagcc gacgcctttg
     2101 acattggctc cttcgatgag gaggacacaa aaggaatcaa gctactggac agtgaccagg
     2161 agctctaccg caacttcccc ctgaccatct cggagcggtg gcagcaggag gtagcagaga
     2221 ctgtctttga caccatcaat gctgagacgg accggctgga ggcccgcaag aaaaccaaaa
     2281 acaagcagtt gggccacgag gaagactacg ccctgggcaa ggactgcatc atgcatggct
     2341 acatgtccaa gatgggcaac cccttcctga cccagtggca gcggcggtac ttctacctgt
     2401 tccctaaccg gctcgagtgg cggggcgagg gcgaggcccc gcagagcctg ctgaccatgg
     2461 aggagatcca gtcggtggag gagacgcaga tcaaggagcg aaagtgcctc ctcctcaaga
     2521 tccgaggtgg caagcagttt gtcctgcagt gcgatagtga cccagagctg gtgcagtgga
     2581 agaaggagct tcgagacgcc taccgcgagg cccagcagct agtgcagcgg gtgcccaaga
     2641 tgaagaacaa gccgcgctcg cccgtcgtgg agctgagcaa ggtgccactg atccagcgcg
     2701 gcagtgccaa cggcctcacc ggtcgccacc atggtgagca agggcgagga gctgttcacc
     2761 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     2821 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagct gatctgcacc
     2881 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta cggcctgcag
     2941 tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     3001 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     3061 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     3121 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     3181 gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac
     3241 aacatcgagg acggcggcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     3301 gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccaa gctgagcaaa
     3361 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     3421 actctcggca tggacgagct gtacaagtaa agcggccgcg actctagatc ataatcagcc
     3481 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     3541 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     3601 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     3661 gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt
     3721 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     3781 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     3841 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     3901 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     3961 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     4021 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     4081 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     4141 ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     4201 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     4261 cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt
     4321 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4381 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     4441 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     4501 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     4561 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     4621 tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc
     4681 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     4741 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     4801 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     4861 caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     4921 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     4981 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     5041 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     5101 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     5161 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac
     5221 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     5281 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     5341 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     5401 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     5461 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     5521 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     5581 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     5641 cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     5701 ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac
     5761 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
     5821 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     5881 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac
     5941 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     6001 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     6061 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     6121 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     6181 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     6241 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     6301 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     6361 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     6421 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     6481 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     6541 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     6601 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     6661 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     6721 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca
     6781 tgcat
//
