LOCUS       sequence-273229-        6797 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 6797)
  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 6797)
  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..6797
                     /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      2677..2686
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2683..2685,2686..2688,2689..3399)
                     /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(2728..2749)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2989..3008)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3336..3357
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3525..3646
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3562..3581)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3616..3635
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3653..4108)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3790..3811)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4002..4021
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4135..4239
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4241..4598
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4241..4432
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4264..4284)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4449..4584
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4633..5427
                     /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             4633..5427
                     /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(4687..4706)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5297..5316
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5615..5634)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5659..5706
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6035..6623
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6524..6543
                     /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 caagcttgcc aagatggccg acctggaggc ggtgctggcc
      601 gacgtgagct acctgatggc catggagaag agcaaggcca cgccggcggc gcgcgccagc
      661 aagaagatcc tgctgcccga gcccagcatc cgcagcgtca tgcagaagta cctggaggac
      721 cggggcgagg tgacttttga gaagatcttc tcccagaagc tggggtacct gcttttccga
      781 gacttctgcc tgaagcacct ggaggaggcc aagcccttgg tagagttcta cgaggagatc
      841 aagaaatacg agaagctgga gacagaggag gagcgcctgg tctgcagccg agagatcttc
      901 gccacgtaca tcatgaagga gctgctggcc tgctcacatc ctttctcgaa gagcgccatt
      961 gagcacgtcc agggccatct ggtgaagaag caggtgcctc cggatctctt ccagccatat
     1021 attgaagaaa tttgccagaa cctccgagga gacgtgttcc agaaattcat cgagagcgat
     1081 aaattcacac gattttgcca gtggaagaat gtagagctca acatccacct gaccatgaac
     1141 gacttcagtg tgcaccgcat catcgggcga ggcggcttcg gtgaggtcta cggctgccgg
     1201 aaggccgaca cgggcaagat gtacgccatg aggtgtctgg acaagaagcg catcaagatg
     1261 aagcaagggg agactctggc cctgaatgag cgcatcatgc tgtcgctcgt cagcaccggg
     1321 gactgcccgt tcatcgtctg catgtcatac gccttccaca caccggacaa gctcagcttc
     1381 atcctggatc tcatgaacgg cggggacctg cactaccacc tgtcccagca cggggtcttc
     1441 tccgaggccg acatgcgttt ctacgccgcc gagatcatcc tgggcctgga gcacatgcac
     1501 aaccgcttcg tggtctaccg ggacctgaag ccggccaaca tcctgctgga cgagcacggc
     1561 cacgtgcgca tctcagacct gggcctggcc tgtgacttct ccaagaagaa gcctcacgcc
     1621 agtgtgggca cccacgggta catggctccc gaggttctac agaagggtgt ggcctacgac
     1681 agcagcgccg actggttctc cctgggctgc atgctcttca agctgctgcg agggcatagc
     1741 cctttccggc agcacaagac caaagacaag catgagatcg acagaatgac attgacaatg
     1801 gctgtggagc tgcctgactc cttctcccct gagctccgct ccttgctgga ggggctgctg
     1861 cagagggatg tcaatcggag gctaggctgc ctgggccgag gggcccagga ggtgaaggag
     1921 agccccttct tccgttccct ggactggcag atggtctttt tacaaaagta ccctcccccg
     1981 ttgatccccc cacgagggga ggtgaatgca gccgacgcct ttgacattgg ctccttcgat
     2041 gaggaggaca caaaaggaat caagctactg gacagtgacc aggagctcta ccgcaacttc
     2101 cccctgacca tctcggagcg gtggcagcag gaggtagcag agactgtctt tgacaccatc
     2161 aatgctgaga cggaccggct ggaggcccgc aagaaaacca aaaacaagca gttgggccac
     2221 gaggaagact acgccctggg caaggactgc atcatgcatg gctacatgtc caagatgggc
     2281 aaccccttcc tgacccagtg gcagcggcgg tacttctacc tgttccctaa ccggctcgag
     2341 tggcggggcg agggcgaggc cccgcagagc ctgctgacca tggaggagat ccagtcggtg
     2401 gaggagacgc agatcaagga gcgaaagtgc ctcctcctca agatccgagg tggcaagcag
     2461 tttgtcctgc agtgcgatag tgacccagag ctggtgcagt ggaagaagga gcttcgagac
     2521 gcctaccgcg aggcccagca gctagtgcag cgggtgccca agatgaagaa caagccgcgc
     2581 tcgcccgtcg tggagctgag caaggtgcca ctgatccagc gtggcagtgc caacggcctc
     2641 tggtcgacgg taccgcgggc ccgggatcca ccggtcgcca ccatggtgag caagggcgag
     2701 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     2761 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     2821 ctgatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgggc
     2881 tacggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag
     2941 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     3001 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     3061 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     3121 aacagccaca acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc
     3181 aagatccgcc acaacatcga ggacggcggc gtgcagctcg ccgaccacta ccagcagaac
     3241 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc
     3301 aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     3361 gccgccggga tcactctcgg catggacgag ctgtacaagt aaagcggccg cgactctaga
     3421 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     3481 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     3541 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     3601 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta
     3661 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     3721 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     3781 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     3841 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     3901 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     3961 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     4021 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     4081 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4141 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4201 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     4261 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4321 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4381 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4441 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4501 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     4561 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
     4621 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     4681 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     4741 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4801 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4861 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4921 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4981 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     5041 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     5101 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     5161 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     5221 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     5281 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     5341 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     5401 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     5461 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     5521 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     5581 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
     5641 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
     5701 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     5761 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     5821 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
     5881 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5941 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     6001 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     6061 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     6121 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6181 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6241 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6301 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6361 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6421 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6481 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6541 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6601 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6661 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     6721 gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc
     6781 ccatatatgg agttccg
//
