LOCUS       sequence-273171-        6164 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Visualization of the Histamine-1 receptor DRY mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6164)
  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 6164)
  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: HsH1R DRY->AAY-mCherry
FEATURES             Location/Qualifiers
     source          1..6164
                     /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      1749..1758
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1755..2465
                     /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(1900..1918)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2168..2187
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2588..2709
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2625..2644)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2679..2698
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2716..3171)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2853..2874)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3065..3084
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3198..3302
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3304..3661
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3304..3495
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3327..3347)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3512..3647
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3696..4490
                     /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             3696..4490
                     /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(3750..3769)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4360..4379
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4678..4697)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4722..4769
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5098..5686
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5587..5606
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5861..6164
                     /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 cttatgagcc tccccaattc ctcctgcctc ttagaagaca
      301 agatgtgtga gggcaacaag accactatgg ccagccccca gctgatgccc ctggtggtgg
      361 tcctgagcac tatctgcttg gtcacagtag ggctcaacct gctggtgctg tatgccgtac
      421 ggagtgagcg gaagctccac actgtgggga acctgtacat cgtcagcctc tcggtggcgg
      481 acttgatcgt gggtgccgtc gtcatgccta tgaacatcct ctacctgctc atgtccaagt
      541 ggtcactggg ccgtcctctc tgcctctttt ggctttccat ggactatgtg gccagcacag
      601 cgtccatttt cagtgtcttc atcctgtgca ttgctgccta ccgctctgtc cagcagcccc
      661 tcaggtacct taagtatcgt accaagaccc gagcctcggc caccattctg ggggcctggt
      721 ttctctcttt tctgtgggtt attcccattc taggctggaa tcacttcatg cagcagacct
      781 cggtgcgccg agaggacaag tgtgagacag acttctatga tgtcacctgg ttcaaggtca
      841 tgactgccat catcaacttc tacctgccca ccttgctcat gctctggttc tatgccaaga
      901 tctacaaggc cgtacgacaa cactgccagc accgggagct catcaatagg tccctccctt
      961 ccttctcaga aattaagctg aggccagaga accccaaggg ggatgccaag aaaccaggga
     1021 aggagtctcc ctgggaggtt ctgaaaagga agccaaaaga tgctggtggt ggatctgtct
     1081 tgaagtcacc atcccaaacc cccaaggaga tgaaatcccc agttgtcttc agccaagagg
     1141 atgatagaga agtagacaaa ctctactgct ttccacttga tattgtgcac atgcaggctg
     1201 cggcagaggg gagtagcagg gactatgtag ccgtcaaccg gagccatggc cagctcaaga
     1261 cagatgagca gggcctgaac acacatgggg ccagcgagat atcagaggat cagatgttag
     1321 gtgatagcca gtccttctct cgaacggact cagataccac cacagagaca gcaccaggca
     1381 aaggcaaatt gaggagtggg tctaacacag gcctggatta catcaagttt acttggaaga
     1441 ggctccgctc gcattcaaga cagtatgtat ctgggttgca catgaaccgc gaaaggaagg
     1501 ccgccaaaca gttgggtttt atcatggcag ccttcatcct ctgctggatc ccttatttca
     1561 tcttcttcat ggtcattgcc ttctgcaaga actgttgcaa tgaacatttg cacatgttca
     1621 ccatctggct gggctacatc aactccacac tgaaccccct catctacccc ttgtgcaatg
     1681 agaacttcaa gaagacattc aagagaattc tgcatattcg ctccccgcgg gcccgggatc
     1741 caccggtcgc caccatggtg agcaagggcg aggaggataa catggccatc atcaaggagt
     1801 tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg
     1861 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg
     1921 gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg
     1981 cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct
     2041 tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact
     2101 cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct
     2161 ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt
     2221 accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg
     2281 gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg
     2341 gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg
     2401 tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca
     2461 agtaaagcgg ccgcgactct agatcataat cagccatacc acatttgtag aggttttact
     2521 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     2581 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     2641 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     2701 tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     2761 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     2821 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     2881 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     2941 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     3001 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     3061 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     3121 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     3181 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     3241 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     3301 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3361 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3421 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3481 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3541 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3601 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3661 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     3721 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     3781 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     3841 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     3901 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     3961 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     4021 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     4081 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     4141 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     4201 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     4261 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     4321 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     4381 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     4441 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     4501 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     4561 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     4621 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga
     4681 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     4741 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     4801 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     4861 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     4921 aggccctgcc atagcctcag gttactcata tatactttag attgatttaa aacttcattt
     4981 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     5041 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     5101 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     5161 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     5221 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     5281 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     5341 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     5401 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     5461 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     5521 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     5581 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     5641 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     5701 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     5761 atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa tagtaatcaa
     5821 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     5881 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     5941 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     6001 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     6061 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     6121 ctacttggca gtacatctac gtattagtca tcgctattac catg
//
