LOCUS       sequence-273174-        6432 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Visualization of GRK6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6432)
  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 6432)
  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: HsGRK6[L66A E514A]-sYFP2
FEATURES             Location/Qualifiers
     source          1..6432
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2008..2017
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2014..2016,2017..2019,2020..2733)
                     /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(2059..2080)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2320..2339)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2667..2688
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2856..2977
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2893..2912)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2947..2966
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2984..3439)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3121..3142)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3333..3352
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3466..3570
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3572..3929
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3572..3763
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3595..3615)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3780..3915
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3964..4758
                     /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             3964..4758
                     /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(4018..4037)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4628..4647
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4946..4965)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4990..5037
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5366..5954
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5855..5874
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        6129..6432
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
      241 actcagatct cgagctcaag cttcgaattc atggagctcg agaacatcgt agcgaacacg
      301 gtgctactca aggcccggga aggtggcggt ggaaatcgca aaggcaaaag caagaaatgg
      361 cggcagatgc tccagttccc tcacatcagc cagtgcgaag agctgcggct cagcctcgag
      421 cgtgactatc acagcctgtg cgagcggcag cccattgggc gcctggcgtt ccgagagttc
      481 tgtgccacga ggccggagct gagccgctgc gtcgccttcc tggatggggt ggccgagtat
      541 gaagtgaccc cggatgacaa gcggaaggca tgtgggcggc agctaacgca gaattttctg
      601 agccacacgg gtcctgacct catccctgag gtcccccggc agctggtgac gaactgcacc
      661 cagcggctgg agcagggtcc ctgcaaagac cttttccagg aactcacccg gctgacccac
      721 gagtacctga gcgtggcccc ttttgccgac tacctcgaca gcatctactt caaccgtttc
      781 ctgcagtgga agtggctgga aaggcagcca gtgaccaaaa acaccttcag gcaataccga
      841 gtcctgggca aaggtggctt tggggaggtg tgcgcctgcc aggtgcgggc cacaggtaag
      901 atgtatgcct gcaagaagct agagaaaaag cggatcaaga agcggaaagg ggaggccatg
      961 gcgctgaacg agaagcagat cctggagaaa gtgaacagta ggtttgtagt gagcttggcc
     1021 tacgcctatg agaccaagga cgcgctgtgc ctggtgctga cactgatgaa cgggggcgac
     1081 ctcaagttcc acatctacca catgggccag gctggcttcc ccgaagcgcg ggccgtcttc
     1141 tacgccgccg agatctgctg tggcctggag gacctgcacc gggagcgcat cgtgtacagg
     1201 gacctgaagc ccgagaacat cttgctggat gaccacggcc acatccgcat ctctgacctg
     1261 ggactagctg tgcatgtgcc cgagggccag accatcaaag ggcgtgtggg caccgtgggt
     1321 tacatggctc cggaggtggt gaagaatgaa cggtacacgt tcagccctga ctggtgggcg
     1381 ctcggctgcc tcctgtacga gatgatcgca ggccagtcgc ccttccagca gaggaagaag
     1441 aagatcaagc gggaggaggt ggagcggctg gtgaaggagg tccccgagga gtattccgag
     1501 cgcttttccc cgcaggcccg ctcactttgc tcacagctcc tctgcaagga ccctgccgaa
     1561 cgcctggggt gtcgtggggg cagtgcccgc gaggtgaagg agcaccccct ctttaagaag
     1621 ctgaacttca agcggctggg agctggcatg ctggagccgc cgttcaagcc tgacccccag
     1681 gccatttact gcaaggatgt tctggacatt gaacagttct ctacggtcaa gggcgtggag
     1741 ctggagccta ccgaccagga cttctaccag aagtttgcca caggcagtgt gcccatcccc
     1801 tggcagaacg cgatggtgga gaccgagtgc ttccaagagc tgaatgtctt tgggctggat
     1861 ggctcagttc ccccagacct ggactggaag ggccagccac ctgcacctcc taaaaaggga
     1921 ctgctgcaga gactcttcag tcgccaagat tgctgtggaa actgcagcga cagcgaggaa
     1981 gagctcccca cccgcctcgg accggtcgcc accatggtga gcaagggcga ggagctgttc
     2041 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     2101 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gctgatctgc
     2161 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctggg ctacggcctg
     2221 cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     2281 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     2341 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     2401 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     2461 aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc
     2521 cacaacatcg aggacggcgg cgtgcagctc gccgaccact accagcagaa cacccccatc
     2581 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc caagctgagc
     2641 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     2701 atcactctcg gcatggacga gctgtacaag taaagcggcc gcgactctag atcataatca
     2761 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     2821 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     2881 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     2941 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat
     3001 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     3061 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     3121 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     3181 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     3241 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     3301 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     3361 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     3421 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3481 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3541 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     3601 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3661 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3721 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3781 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3841 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     3901 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     3961 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     4021 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     4081 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     4141 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     4201 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     4261 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     4321 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     4381 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     4441 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     4501 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     4561 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     4621 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     4681 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     4741 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     4801 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     4861 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     4921 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     4981 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     5041 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     5101 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     5161 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     5221 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     5281 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     5341 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     5401 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     5461 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     5521 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     5581 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     5641 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     5701 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     5761 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     5821 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     5881 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5941 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     6001 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     6061 ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     6121 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     6181 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     6241 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     6301 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     6361 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     6421 gctattacca tg
//
