LOCUS       sequence-273122-        6230 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 6230)
  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 6230)
  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-p2A-mCherry
FEATURES             Location/Qualifiers
     source          1..6230
                     /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"
     CDS             2062..2118
                     /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             2125..2835
                     /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(2270..2288)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2538..2557
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2958..3079
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2995..3014)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3049..3068
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3086..3541)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3223..3244)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3435..3454
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3568..3672
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3674..4031
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3674..3865
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3697..3717)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3882..4017
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4066..4860
                     /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             4066..4860
                     /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(4120..4139)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4730..4749
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5048..5067)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5092..5139
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5468..6056
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5957..5976
                     /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 caagcttatg agcctcccca attcctcctg cctcttagaa
      601 gacaagatgt gtgagggcaa caagaccact atggccagcc cccagctgat gcccctggtg
      661 gtggtcctga gcactatctg cttggtcaca gtagggctca acctgctggt gctgtatgcc
      721 gtacggagtg agcggaagct ccacactgtg gggaacctgt acatcgtcag cctctcggtg
      781 gcggacttga tcgtgggtgc cgtcgtcatg cctatgaaca tcctctacct gctcatgtcc
      841 aagtggtcac tgggccgtcc tctctgcctc ttttggcttt ccatggacta tgtggccagc
      901 acagcgtcca ttttcagtgt cttcatcctg tgcattgctg cctaccgctc tgtccagcag
      961 cccctcaggt accttaagta tcgtaccaag acccgagcct cggccaccat tctgggggcc
     1021 tggtttctct cttttctgtg ggttattccc attctaggct ggaatcactt catgcagcag
     1081 acctcggtgc gccgagagga caagtgtgag acagacttct atgatgtcac ctggttcaag
     1141 gtcatgactg ccatcatcaa cttctacctg cccaccttgc tcatgctctg gttctatgcc
     1201 aagatctaca aggccgtacg acaacactgc cagcaccggg agctcatcaa taggtccctc
     1261 ccttccttct cagaaattaa gctgaggcca gagaacccca agggggatgc caagaaacca
     1321 gggaaggagt ctccctggga ggttctgaaa aggaagccaa aagatgctgg tggtggatct
     1381 gtcttgaagt caccatccca aacccccaag gagatgaaat ccccagttgt cttcagccaa
     1441 gaggatgata gagaagtaga caaactctac tgctttccac ttgatattgt gcacatgcag
     1501 gctgcggcag aggggagtag cagggactat gtagccgtca accggagcca tggccagctc
     1561 aagacagatg agcagggcct gaacacacat ggggccagcg agatatcaga ggatcagatg
     1621 ttaggtgata gccagtcctt ctctcgaacg gactcagata ccaccacaga gacagcacca
     1681 ggcaaaggca aattgaggag tgggtctaac acaggcctgg attacatcaa gtttacttgg
     1741 aagaggctcc gctcgcattc aagacagtat gtatctgggt tgcacatgaa ccgcgaaagg
     1801 aaggccgcca aacagttggg ttttatcatg gcagccttca tcctctgctg gatcccttat
     1861 ttcatcttct tcatggtcat tgccttctgc aagaactgtt gcaatgaaca tttgcacatg
     1921 ttcaccatct ggctgggcta catcaactcc acactgaacc ccctcatcta ccccttgtgc
     1981 aatgagaact tcaagaagac attcaagaga attctgcata ttcgctcccc gcgggcccgg
     2041 gatccaccgg tgggatcggg tgctactaac ttcagcctgc tgaagcaggc tggagacgtg
     2101 gaggagaacc ctggacctgg gtccatggtg agcaagggcg aggaggataa catggccatc
     2161 atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc
     2221 gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag
     2281 gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac
     2341 ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc
     2401 cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg
     2461 acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc
     2521 aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc
     2581 gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg
     2641 aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg
     2701 cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac
     2761 tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac
     2821 gagctgtaca agtaaagcgg ccgcgactct agatcataat cagccatacc acatttgtag
     2881 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     2941 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     3001 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     3061 aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     3121 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     3181 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     3241 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     3301 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     3361 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     3421 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     3481 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     3541 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     3601 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     3661 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     3721 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3781 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3841 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     3901 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3961 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4021 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     4081 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4141 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4201 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     4261 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4321 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4381 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4441 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4501 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4561 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     4621 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4681 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4741 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4801 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4861 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     4921 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     4981 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     5041 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     5101 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     5161 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     5221 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     5281 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     5341 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     5401 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     5461 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     5521 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     5581 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     5641 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     5701 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     5761 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     5821 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     5881 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     5941 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     6001 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     6061 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     6121 ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa
     6181 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
//
