LOCUS       sequence-160921-        6067 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Histamine-3 receptor tagged with mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6067)
  AUTHORS   van Unen J, Rashidfarrokhi A, Hoogendoorn E, Postma M, Gadella TW 
            Jr, Goedhart J
  TITLE     Quantitative Single-Cell Analysis of Signaling Pathways Activated 
            Immediately Downstream of Histamine Receptor Subtypes.
  JOURNAL   Mol Pharmacol. 2016 Sep;90(3):162-76. doi: 10.1124/mol.116.104505. 
            Epub 2016 Jun 29.
  PUBMED    27358232
REFERENCE   2  (bases 1 to 6067)
  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: 
            "10.1124/mol.116.104505"; journalName: "Mol Pharmacol"; date: 
            "2016-09"; volume: "90"; issue: "3"; pages: "162-76"
COMMENT     Sequence Label: pH3R-mCherry-N1
FEATURES             Location/Qualifiers
     source          1..6067
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2016..2025
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2022..2732
                     /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(2167..2185)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2435..2454
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2855..2976
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2892..2911)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2946..2965
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2983..3438)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3120..3141)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3332..3351
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3465..3569
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3571..3928
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3571..3762
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3594..3614)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3779..3914
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3963..4757
                     /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             3963..4757
                     /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(4017..4036)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4627..4646
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4945..4964)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4989..5036
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5365..5953
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5854..5873
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttggt accgagctcg gatccactag tccagtgtgg
      661 tggaattcac catggagcgc gcgccgcccg acgggccgct gaacgcttcg ggggcgctgg
      721 cgggcgatgc ggcggcggcg ggcggggcgc gcggcttctc ggcagcctgg accgcggtgc
      781 tggccgcgct catggcgctg ctcatcgtgg ccacggtgct gggcaacgcg ctggtcatgc
      841 tcgccttcgt ggccgactcg agcctccgca cccagaacaa cttcttcctg ctcaacctcg
      901 ccatctccga cttcctcgtc ggcgccttct gcatcccact gtatgtaccc tacgtgctga
      961 caggccgctg gaccttcggc cggggcctct gcaagctgtg gctggtagtg gactacctgc
     1021 tgtgcacctc ctctgccttc aacatcgtgc tcatcagcta cgaccgcttc ctgtcggtca
     1081 cccgagcggt ctcataccgg gcccagcagg gtgacacgcg gcgggcagtg cggaagatgc
     1141 tgctggtgtg ggtgctggcc ttcctgctgt acggaccagc catcctgagc tgggagtacc
     1201 tgtccggggg cagctccatc cccgagggcc actgctatgc cgagttcttc tacaactggt
     1261 acttcctcat cacggcttcc accctggagt tctttacgcc cttcctcagc gtcaccttct
     1321 ttaacctcag catctacctg aacatccaga ggcgcacccg cctccggctg gatggggctc
     1381 gagaggcagc cggccccgag ccccctcccg aggcccagcc ctcaccaccc ccaccgcctg
     1441 gctgctgggg ctgctggcag aaggggcacg gggaggccat gccgctgcac aggtatgggg
     1501 tgggtgaggc ggccgtaggc gctgaggccg gggaggcgac cctcgggggt ggcggtgggg
     1561 gcggctccgt ggcttcaccc acctccagct ccggcagctc ctcgaggggc actgagaggc
     1621 cgcgctcact caagaggggc tccaagccgt cggcgtcctc ggcctcgctg gagaagcgca
     1681 tgaagatggt gtcccagagc ttcacccagc gctttcggct gtctcgggac aggaaagtgg
     1741 ccaagtcgct ggccgtcatc gtgagcatct ttgggctctg ctgggcccca tacacgctgc
     1801 tgatgatcat ccgggccgcc tgccatggcc actgcgtccc tgactactgg tacgaaacct
     1861 ccttctggct cctgtgggcc aactcggctg tcaaccctgt cctctaccct ctgtgccacc
     1921 acagcttccg ccgggccttc accaagctgc tctgccccca gaagctcaaa atccagcccc
     1981 acagctccct ggagcactgc tggaagggac cggtcgccac catggtgagc aagggcgagg
     2041 aggataacat ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg
     2101 tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc
     2161 agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt
     2221 cccctcagtt catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact
     2281 acttgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg
     2341 gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg
     2401 tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag aagaccatgg
     2461 gctgggaggc ctcctccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca
     2521 agcagaggct gaagctgaag gacggcggcc actacgacgc tgaggtcaag accacctaca
     2581 aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt caacatcaag ttggacatca
     2641 cctcccacaa cgaggactac accatcgtgg aacagtacga acgcgccgag ggccgccact
     2701 ccaccggcgg catggacgag ctgtacaagt aaagcggccg cgactctaga tcataatcag
     2761 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     2821 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     2881 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     2941 tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata
     3001 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     3061 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     3121 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     3181 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     3241 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     3301 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     3361 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     3421 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     3481 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     3541 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
     3601 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3661 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     3721 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     3781 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     3841 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     3901 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
     3961 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4021 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4081 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4141 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4201 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     4261 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     4321 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     4381 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     4441 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     4501 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     4561 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     4621 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     4681 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     4741 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     4801 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     4861 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     4921 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     4981 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
     5041 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     5101 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     5161 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
     5221 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     5281 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     5341 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     5401 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     5461 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     5521 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     5581 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     5641 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     5701 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     5761 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     5821 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     5881 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     5941 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     6001 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     6061 catgcat
//
