LOCUS       sequence-273233-        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-mVenus
FEATURES             Location/Qualifiers
     source          1..6432
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2312..2321
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2318..2320,2321..2323,2324..3034)
                     /codon_start=1
                     /product="monomeric optimized variant of EYFP, identical to
                     mVenus but with L68V (Kremers et al., 2006)"
                     /label=SYFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2363..2384)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2624..2643)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2971..2992
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3160..3281
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3197..3216)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3251..3270
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3288..3743)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3425..3446)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3637..3656
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3770..3874
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3876..4233
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3876..4067
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3899..3919)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4084..4219
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4268..5062
                     /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             4268..5062
                     /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(4322..4341)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4932..4951
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5250..5269)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5294..5341
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5670..6258
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6159..6178
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagct caagcttcga attcatggag ctcgagaaca tcgtagcgaa
      601 cacggtgcta ctcaaggccc gggaaggtgg cggtggaaat cgcaaaggca aaagcaagaa
      661 atggcggcag atgctccagt tccctcacat cagccagtgc gaagagctgc ggctcagcct
      721 cgagcgtgac tatcacagcc tgtgcgagcg gcagcccatt gggcgcctgc tgttccgaga
      781 gttctgtgcc acgaggccgg agctgagccg ctgcgtcgcc ttcctggatg gggtggccga
      841 gtatgaagtg accccggatg acaagcggaa ggcatgtggg cggcagctaa cgcagaattt
      901 tctgagccac acgggtcctg acctcatccc tgaggtcccc cggcagctgg tgacgaactg
      961 cacccagcgg ctggagcagg gtccctgcaa agaccttttc caggaactca cccggctgac
     1021 ccacgagtac ctgagcgtgg ccccttttgc cgactacctc gacagcatct acttcaaccg
     1081 tttcctgcag tggaagtggc tggaaaggca gccagtgacc aaaaacacct tcaggcaata
     1141 ccgagtcctg ggcaaaggtg gctttgggga ggtgtgcgcc tgccaggtgc gggccacagg
     1201 taagatgtat gcctgcaaga agctagagaa aaagcggatc aagaagcgga aaggggaggc
     1261 catggcgctg aacgagaagc agatcctgga gaaagtgaac agtaggtttg tagtgagctt
     1321 ggcctacgcc tatgagacca aggacgcgct gtgcctggtg ctgacactga tgaacggggg
     1381 cgacctcaag ttccacatct accacatggg ccaggctggc ttccccgaag cgcgggccgt
     1441 cttctacgcc gccgagatct gctgtggcct ggaggacctg caccgggagc gcatcgtgta
     1501 cagggacctg aagcccgaga acatcttgct ggatgaccac ggccacatcc gcatctctga
     1561 cctgggacta gctgtgcatg tgcccgaggg ccagaccatc aaagggcgtg tgggcaccgt
     1621 gggttacatg gctccggagg tggtgaagaa tgaacggtac acgttcagcc ctgactggtg
     1681 ggcgctcggc tgcctcctgt acgagatgat cgcaggccag tcgcccttcc agcagaggaa
     1741 gaagaagatc aagcgggagg aggtggagcg gctggtgaag gaggtccccg aggagtattc
     1801 cgagcgcttt tccccgcagg cccgctcact ttgctcacag ctcctctgca aggaccctgc
     1861 cgaacgcctg gggtgtcgtg ggggcagtgc ccgcgaggtg aaggagcacc ccctctttaa
     1921 gaagctgaac ttcaagcggc tgggagctgg catgctggag ccgccgttca agcctgaccc
     1981 ccaggccatt tactgcaagg atgttctgga cattgaacag ttctctacgg tcaagggcgt
     2041 ggagctggag cctaccgacc aggacttcta ccagaagttt gccacaggca gtgtgcccat
     2101 cccctggcag aacgagatgg tggagaccga gtgcttccaa gagctgaatg tctttgggct
     2161 ggatggctca gttcccccag acctggactg gaagggccag ccacctgcac ctcctaaaaa
     2221 gggactgctg cagagactct tcagtcgcca agattgctgt ggaaactgca gcgacagcga
     2281 ggaagagctc cccacccgcc tcggaccggt cgccaccatg gtgagcaagg gcgaggagct
     2341 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     2401 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagctgat
     2461 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgggctacgg
     2521 cgtgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     2581 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     2641 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     2701 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     2761 ccacaacgtc tatatcaccg ccgacaagca gaagaacggc atcaaggcca acttcaagat
     2821 ccgccacaac atcgaggacg gcggcgtgca gctcgccgac cactaccagc agaacacccc
     2881 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccaagct
     2941 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     3001 cgggatcact ctcggcatgg acgagctgta caagtaaagc ggccgcgact ctagatcata
     3061 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     3121 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     3181 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     3241 cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt
     3301 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     3361 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     3421 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     3481 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     3541 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     3601 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     3661 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     3721 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     3781 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3841 ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg
     3901 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3961 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     4021 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     4081 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     4141 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     4201 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc
     4261 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     4321 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     4381 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     4441 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     4501 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     4561 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     4621 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     4681 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     4741 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
     4801 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     4861 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     4921 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     4981 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     5041 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     5101 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     5161 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     5221 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga
     5281 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt
     5341 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     5401 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     5461 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
     5521 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     5581 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5641 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5701 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5761 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     5821 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     5881 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     5941 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     6001 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     6061 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     6121 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     6181 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6241 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6301 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     6361 accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     6421 tatggagttc cg
//
