LOCUS       sequence-273118-        5832 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Visualization of the At1aR DRY mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5832)
  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 5832)
  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: RnAT1aR DRY->AAY-p2A-mCherry
FEATURES             Location/Qualifiers
     source          1..5832
                     /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"
     CDS             1360..1416
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1423..2133
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1568..1586)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1836..1855
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2256..2377
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2293..2312)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2347..2366
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2384..2839)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2521..2542)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2733..2752
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2866..2970
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2972..3329
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2972..3163
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2995..3015)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3180..3315
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3364..4158
                     /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             3364..4158
                     /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(3418..3437)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4028..4047
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4346..4365)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4390..4437
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4766..5354
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5255..5274
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5529..5832
                     /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 cttcaaaatg gcccttaact cttctgctga agatggtatc
      301 aaaagaatcc aagatgactg ccccaaggct ggcaggcaca gttacatatt tgtcatgatc
      361 cctaccctct acagcatcat ctttgtggtg ggaatatttg gaaacagctt ggtggtgatt
      421 gtcatttact tttacatgaa gctgaagact gtggccagcg tctttcttct caatctcgcc
      481 ttggctgact tatgcttttt gctgactttg cccctgtggg cagtctatac cgctatggag
      541 taccgctggc ccttcggcaa tcacctatgt aagatcgctt cggccagcgt gagcttcaac
      601 ctctacgcca gtgtgttcct tctcacgtgt ctcagcatcg ccgcctacct ggccatcgtc
      661 cacccaatga agtctcgcct tcgccgcacg atgctggtgg ccaaagtcac ctgcatcatc
      721 atctggctga tggctggctt ggccagtttg ccagctgtca tccaccgaaa tgtatacttc
      781 atcgagaaca ccaatatcac agtgtgcgcg tttcattatg agtctcggaa ttcgacgctc
      841 cccatagggc tgggccttac caagaatatt ctgggcttct tgttcccttt ccttatcatt
      901 ctcaccagct atacccttat ttggaaagct ctaaagaagg cttatgaaat tcaaaagaac
      961 aaaccaagaa acgatgacat ctttaggata attatggcga ttgtgctttt cttcttcttt
     1021 tcctgggtcc cccaccaaat attcactttc ctggatgtgc tgattcagct gggcgtcatc
     1081 catgactgta aaatttctga catcgtggac actgccatgc ccatcaccat ctgcatagcg
     1141 tattttaaca actgcctgaa ccctctgttc tacggctttc tggggaagaa atttaaaaag
     1201 tatttcctcc agctcctgaa atatattccc ccaaaggcca agtcccactc aagcctgtct
     1261 acgaaaatga gcacgctttc ttaccggcct tcggataaca tgagctcatc ggccaaaaag
     1321 cctgcgtctt gttttgaggt ggaaccggtg ggatcgggtg ctactaactt cagcctgctg
     1381 aagcaggctg gagacgtgga ggagaaccct ggacctgggt ccatggtgag caagggcgag
     1441 gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc
     1501 gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc
     1561 cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg
     1621 tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac
     1681 tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac
     1741 ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag
     1801 gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg
     1861 ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc
     1921 aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac
     1981 aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gttggacatc
     2041 acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac
     2101 tccaccggcg gcatggacga gctgtacaag taaagcggcc gcgactctag atcataatca
     2161 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     2221 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     2281 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     2341 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat
     2401 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     2461 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     2521 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     2581 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     2641 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     2701 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     2761 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     2821 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     2881 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     2941 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     3001 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3061 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3121 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3181 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3241 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     3301 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     3361 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     3421 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     3481 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     3541 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     3601 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     3661 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     3721 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     3781 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     3841 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     3901 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     3961 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     4021 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     4081 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     4141 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     4201 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     4261 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     4321 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     4381 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     4441 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     4501 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     4561 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     4621 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4681 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4741 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4801 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4861 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     4921 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4981 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     5041 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     5101 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     5161 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     5221 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     5281 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5341 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     5401 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     5461 ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     5521 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     5581 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     5641 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     5701 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     5761 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     5821 gctattacca tg
//
