LOCUS       sequence-160923-        6164 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Histamine-1 receptor tagged with mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6164)
  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 6164)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: pH1R-mCherry-N1
FEATURES             Location/Qualifiers
     source          1..6164
                     /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      2113..2122
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2119..2829
                     /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(2264..2282)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2532..2551
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2952..3073
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2989..3008)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3043..3062
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3080..3535)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3217..3238)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3429..3448
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3562..3666
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3668..4025
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3668..3859
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3691..3711)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3876..4011
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4060..4854
                     /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             4060..4854
                     /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(4114..4133)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4724..4743
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5042..5061)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5086..5133
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5462..6050
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5951..5970
                     /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 caagcttatg agcctcccca attcctcctg cctcttagaa
      661 gacaagatgt gtgagggcaa caagaccact atggccagcc cccagctgat gcccctggtg
      721 gtggtcctga gcactatctg cttggtcaca gtagggctca acctgctggt gctgtatgcc
      781 gtacggagtg agcggaagct ccacactgtg gggaacctgt acatcgtcag cctctcggtg
      841 gcggacttga tcgtgggtgc cgtcgtcatg cctatgaaca tcctctacct gctcatgtcc
      901 aagtggtcac tgggccgtcc tctctgcctc ttttggcttt ccatggacta tgtggccagc
      961 acagcgtcca ttttcagtgt cttcatcctg tgcattgatc gctaccgctc tgtccagcag
     1021 cccctcaggt accttaagta tcgtaccaag acccgagcct cggccaccat tctgggggcc
     1081 tggtttctct cttttctgtg ggttattccc attctaggct ggaatcactt catgcagcag
     1141 acctcggtgc gccgagagga caagtgtgag acagacttct atgatgtcac ctggttcaag
     1201 gtcatgactg ccatcatcaa cttctacctg cccaccttgc tcatgctctg gttctatgcc
     1261 aagatctaca aggccgtacg acaacactgc cagcaccggg agctcatcaa taggtccctc
     1321 ccttccttct cagaaattaa gctgaggcca gagaacccca agggggatgc caagaaacca
     1381 gggaaggagt ctccctggga ggttctgaaa aggaagccaa aagatgctgg tggtggatct
     1441 gtcttgaagt caccatccca aacccccaag gagatgaaat ccccagttgt cttcagccaa
     1501 gaggatgata gagaagtaga caaactctac tgctttccac ttgatattgt gcacatgcag
     1561 gctgcggcag aggggagtag cagggactat gtagccgtca accggagcca tggccagctc
     1621 aagacagatg agcagggcct gaacacacat ggggccagcg agatatcaga ggatcagatg
     1681 ttaggtgata gccagtcctt ctctcgaacg gactcagata ccaccacaga gacagcacca
     1741 ggcaaaggca aattgaggag tgggtctaac acaggcctgg attacatcaa gtttacttgg
     1801 aagaggctcc gctcgcattc aagacagtat gtatctgggt tgcacatgaa ccgcgaaagg
     1861 aaggccgcca aacagttggg ttttatcatg gcagccttca tcctctgctg gatcccttat
     1921 ttcatcttct tcatggtcat tgccttctgc aagaactgtt gcaatgaaca tttgcacatg
     1981 ttcaccatct ggctgggcta catcaactcc acactgaacc ccctcatcta ccccttgtgc
     2041 aatgagaact tcaagaagac attcaagaga attctgcata ttcgctcccc gcgggcccgg
     2101 gatccaccgg tcgccaccat ggtgagcaag ggcgaggagg ataacatggc catcatcaag
     2161 gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc
     2221 gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc
     2281 aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc
     2341 aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag
     2401 ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag
     2461 gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc
     2521 ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg
     2581 atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac
     2641 ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg
     2701 cccggcgcct acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc
     2761 atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg
     2821 tacaagtaaa gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt
     2881 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     2941 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     3001 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     3061 tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     3121 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     3181 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     3241 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     3301 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     3361 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     3421 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     3481 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt
     3541 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     3601 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     3661 aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     3721 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3781 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3841 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3901 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3961 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     4021 gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     4081 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     4141 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     4201 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta
     4261 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     4321 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     4381 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     4441 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     4501 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     4561 gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc
     4621 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     4681 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     4741 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     4801 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     4861 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     4921 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     4981 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa
     5041 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     5101 agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct
     5161 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
     5221 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
     5281 cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc
     5341 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     5401 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     5461 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     5521 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     5581 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     5641 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     5701 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     5761 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     5821 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     5881 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     5941 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     6001 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     6061 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     6121 cgttatcccc tgattctgtg gataaccgta ttaccgccat gcat
//
