LOCUS       sequence-266023-        6398 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 6398)
  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 6398)
  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..6398
                     /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      2338..2347
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2344..2346,2347..2349,2350..3063)
                     /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(2389..2410)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2650..2669)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2997..3018
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3186..3307
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3223..3242)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3277..3296
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3314..3769)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3451..3472)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3663..3682
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3796..3900
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3902..4259
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3902..4093
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3925..3945)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4110..4245
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4294..5088
                     /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             4294..5088
                     /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(4348..4367)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4958..4977
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5276..5295)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5320..5367
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5696..6284
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6185..6204
                     /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 atggagctcg
      661 agaacatcgt agcgaacacg gtgctactca aggcccggga aggtggcggt ggaaatcgca
      721 aaggcaaaag caagaaatgg cggcagatgc tccagttccc tcacatcagc cagtgcgaag
      781 agctgcggct cagcctcgag cgtgactatc acagcctgtg cgagcggcag cccattgggc
      841 gcctggcgtt ccgagagttc tgtgccacga ggccggagct gagccgctgc gtcgccttcc
      901 tggatggggt ggccgagtat gaagtgaccc cggatgacaa gcggaaggca tgtgggcggc
      961 agctaacgca gaattttctg agccacacgg gtcctgacct catccctgag gtcccccggc
     1021 agctggtgac gaactgcacc cagcggctgg agcagggtcc ctgcaaagac cttttccagg
     1081 aactcacccg gctgacccac gagtacctga gcgtggcccc ttttgccgac tacctcgaca
     1141 gcatctactt caaccgtttc ctgcagtgga agtggctgga aaggcagcca gtgaccaaaa
     1201 acaccttcag gcaataccga gtcctgggca aaggtggctt tggggaggtg tgcgcctgcc
     1261 aggtgcgggc cacaggtaag atgtatgcct gcaagaagct agagaaaaag cggatcaaga
     1321 agcggaaagg ggaggccatg gcgctgaacg agaagcagat cctggagaaa gtgaacagta
     1381 ggtttgtagt gagcttggcc tacgcctatg agaccaagga cgcgctgtgc ctggtgctga
     1441 cactgatgaa cgggggcgac ctcaagttcc acatctacca catgggccag gctggcttcc
     1501 ccgaagcgcg ggccgtcttc tacgccgccg agatctgctg tggcctggag gacctgcacc
     1561 gggagcgcat cgtgtacagg gacctgaagc ccgagaacat cttgctggat gaccacggcc
     1621 acatccgcat ctctgacctg ggactagctg tgcatgtgcc cgagggccag accatcaaag
     1681 ggcgtgtggg caccgtgggt tacatggctc cggaggtggt gaagaatgaa cggtacacgt
     1741 tcagccctga ctggtgggcg ctcggctgcc tcctgtacga gatgatcgca ggccagtcgc
     1801 ccttccagca gaggaagaag aagatcaagc gggaggaggt ggagcggctg gtgaaggagg
     1861 tccccgagga gtattccgag cgcttttccc cgcaggcccg ctcactttgc tcacagctcc
     1921 tctgcaagga ccctgccgaa cgcctggggt gtcgtggggg cagtgcccgc gaggtgaagg
     1981 agcaccccct ctttaagaag ctgaacttca agcggctggg agctggcatg ctggagccgc
     2041 cgttcaagcc tgacccccag gccatttact gcaaggatgt tctggacatt gaacagttct
     2101 ctacggtcaa gggcgtggag ctggagccta ccgaccagga cttctaccag aagtttgcca
     2161 caggcagtgt gcccatcccc tggcagaacg cgatggtgga gaccgagtgc ttccaagagc
     2221 tgaatgtctt tgggctggat ggctcagttc ccccagacct ggactggaag ggccagccac
     2281 ctgcacctcc taaaaaggga ctgctgcaga gactcttcag tcgccaaagg accggtcgcc
     2341 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     2401 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     2461 tacggcaagc tgaccctgaa gctgatctgc accaccggca agctgcccgt gccctggccc
     2521 accctcgtga ccaccctggg ctacggcctg cagtgcttcg cccgctaccc cgaccacatg
     2581 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     2641 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     2701 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     2761 cacaagctgg agtacaacta caacagccac aacgtctata tcaccgccga caagcagaag
     2821 aacggcatca aggccaactt caagatccgc cacaacatcg aggacggcgg cgtgcagctc
     2881 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     2941 cactacctga gctaccagtc caagctgagc aaagacccca acgagaagcg cgatcacatg
     3001 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     3061 taaagcggcc gcgactctag atcataatca gccataccac atttgtagag gttttacttg
     3121 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     3181 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     3241 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     3301 tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     3361 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     3421 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     3481 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     3541 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     3601 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     3661 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     3721 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact
     3781 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     3841 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     3901 cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     3961 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     4021 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4081 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     4141 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     4201 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag
     4261 atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     4321 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     4381 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     4441 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg
     4501 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     4561 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     4621 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     4681 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     4741 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     4801 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc
     4861 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     4921 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     4981 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     5041 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     5101 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     5161 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     5221 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa
     5281 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     5341 aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     5401 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     5461 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     5521 gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt
     5581 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     5641 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     5701 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     5761 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     5821 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     5881 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     5941 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     6001 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     6061 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     6121 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     6181 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     6241 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     6301 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     6361 cccctgattc tgtggataac cgtattaccg ccatgcat
//
