LOCUS       sequence-160919-        6133 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Co-expresses (untagged) Histamine-3 receptor and mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6133)
  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 6133)
  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-P2A-mCherry-N1
FEATURES             Location/Qualifiers
     source          1..6133
                     /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             2025..2081
                     /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             2088..2798
                     /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(2233..2251)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2501..2520
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2921..3042
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2958..2977)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3012..3031
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3049..3504)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3186..3207)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3398..3417
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3531..3635
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3637..3994
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3637..3828
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3660..3680)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3845..3980
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4029..4823
                     /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             4029..4823
                     /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(4083..4102)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4693..4712
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5011..5030)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5055..5102
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5431..6019
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5920..5939
                     /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 cggtgggatc gggtgctact aacttcagcc
     2041 tgctgaagca ggctggagac gtggaggaga accctggacc tgggtccatg gtgagcaagg
     2101 gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg
     2161 gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg
     2221 gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca
     2281 tcctgtcccc tcagttcatg tacggctcca aggcctacgt gaagcacccc gccgacatcc
     2341 ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg
     2401 aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct
     2461 acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga
     2521 ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg
     2581 agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgctgag gtcaagacca
     2641 cctacaaggc caagaagccc gtgcagctgc ccggcgccta caacgtcaac atcaagttgg
     2701 acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc
     2761 gccactccac cggcggcatg gacgagctgt acaagtaaag cggccgcgac tctagatcat
     2821 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     2881 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     2941 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     3001 gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg
     3061 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     3121 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     3181 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     3241 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     3301 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     3361 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     3421 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     3481 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3541 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     3601 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
     3661 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3721 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     3781 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3841 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3901 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     3961 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
     4021 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     4081 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     4141 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     4201 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     4261 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     4321 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     4381 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     4441 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     4501 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     4561 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     4621 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     4681 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     4741 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     4801 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     4861 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     4921 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     4981 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     5041 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
     5101 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     5161 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     5221 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
     5281 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     5341 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     5401 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5461 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5521 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     5581 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5641 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5701 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     5761 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     5821 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     5881 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     5941 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6001 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6061 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     6121 taccgccatg cat
//
