LOCUS       sequence-160917-        5962 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Co-expresses (untagged) Histamine-4 receptor and mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5962)
  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 5962)
  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: pH4R-P2A-mCherry-N1
FEATURES             Location/Qualifiers
     source          1..5962
                     /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"
     CDS             1854..1910
                     /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             1917..2627
                     /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(2062..2080)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2330..2349
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2750..2871
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2787..2806)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2841..2860
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2878..3333)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3015..3036)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3227..3246
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3360..3464
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3466..3823
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3466..3657
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3489..3509)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3674..3809
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3858..4652
                     /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             3858..4652
                     /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(3912..3931)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4522..4541
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4840..4859)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4884..4931
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5260..5848
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5749..5768
                     /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 caagcttcga attctgcagt cgacggtacc gagctcggat
      661 ccaccatgcc agatactaat agcacaatca atttatcact aagcactcgt gttactttag
      721 cattttttat gtccttagta gcttttgcta taatgctagg aaatgctttg gtcattttag
      781 cttttgtggt ggacaaaaac cttagacatc gaagtagtta tttttttctt aacttggcca
      841 tctctgactt ctttgtgggt gtgatctcca ttcctttgta catccctcac acgctgttcg
      901 aatgggattt tggaaaggaa atctgtgtat tttggctcac tactgactat ctgttatgta
      961 cagcatctgt atataacatt gtcctcatca gctatgatcg atacctgtca gtctcaaatg
     1021 ctgtgtctta tagaactcaa catactgggg tcttgaagat tgttactctg atggtggccg
     1081 tttgggtgct ggccttctta gtgaatgggc caatgattct agtttcagag tcttggaagg
     1141 atgaaggtag tgaatgtgaa cctggatttt tttcggaatg gtacatcctt gccatcacat
     1201 cattcttgga attcgtgatc ccagtcatct tagtcgctta tttcaacatg aatatttatt
     1261 ggagcctgtg gaagcgtgat catctcagta ggtgccaaag ccatcctgga ctgactgctg
     1321 tctcttccaa catctgtgga cactcattca gaggtagact atcttcaagg agatctcttt
     1381 ctgcatcgac agaagttcct gcatcctttc attcagagag acagaggaga aagagtagtc
     1441 tcatgttttc ctcaagaacc aagatgaata gcaatacaat tgcttccaaa atgggttcct
     1501 tctcccaatc agattctgta gctcttcacc aaagggaaca tgttgaactg cttagagcca
     1561 ggagattagc caagtcactg gccattctct taggggtttt tgctgtttgc tgggctccat
     1621 attctctgtt cacaattgtc ctttcatttt attcctcagc aacaggtcct aaatcagttt
     1681 ggtatagaat tgcattttgg cttcagtggt tcaattcctt tgtcaatcct cttttgtatc
     1741 cattgtgtca caagcgcttt caaaaggctt tcttgaaaat attttgtata aaaaagcaac
     1801 ctctaccatc acaacacagt cggtcagtat cttctacacc ggtgggatcg ggtgctacta
     1861 acttcagcct gctgaagcag gctggagacg tggaggagaa ccctggacct gggtccatgg
     1921 tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc
     1981 acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
     2041 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg
     2101 cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg
     2161 ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga
     2221 tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg
     2281 agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc
     2341 agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc
     2401 tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg
     2461 tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca
     2521 tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg
     2581 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtaaagc ggccgcgact
     2641 ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     2701 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     2761 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     2821 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa
     2881 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     2941 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     3001 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     3061 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     3121 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     3181 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     3241 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     3301 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc
     3361 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     3421 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa
     3481 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     3541 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     3601 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     3661 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     3721 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     3781 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag
     3841 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     3901 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     3961 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     4021 gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     4081 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     4141 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     4201 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     4261 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     4321 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     4381 aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     4441 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     4501 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     4561 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     4621 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     4681 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     4741 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     4801 catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat
     4861 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg
     4921 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     4981 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     5041 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc
     5101 aggttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     5161 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     5221 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     5281 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     5341 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     5401 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     5461 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     5521 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     5581 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     5641 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     5701 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     5761 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     5821 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     5881 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     5941 taaccgtatt accgccatgc at
//
