LOCUS       sequence-273185-        6173 bp    DNA     circular SYN 15-APR-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 6173)
  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 6173)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-mTurquoise2
FEATURES             Location/Qualifiers
     source          1..6173
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1749..1758
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1755..1757,1758..1760,1761..2474)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1800..1821)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     2408..2429
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2597..2718
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2634..2653)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2688..2707
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2725..3180)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2862..2883)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3074..3093
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3207..3311
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3313..3670
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3313..3504
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3336..3356)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3521..3656
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3705..4499
                     /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             3705..4499
                     /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(3759..3778)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4369..4388
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4687..4706)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4731..4778
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5107..5695
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5596..5615
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5870..6173
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
      241 actcagatct cgagctcaag cttatgagcc tccccaattc ctcctgcctc ttagaagaca
      301 agatgtgtga gggcaacaag accactatgg ccagccccca gctgatgccc ctggtggtgg
      361 tcctgagcac tatctgcttg gtcacagtag ggctcaacct gctggtgctg tatgccgtac
      421 ggagtgagcg gaagctccac actgtgggga acctgtacat cgtcagcctc tcggtggcgg
      481 acttgatcgt gggtgccgtc gtcatgccta tgaacatcct ctacctgctc atgtccaagt
      541 ggtcactggg ccgtcctctc tgcctctttt ggctttccat ggactatgtg gccagcacag
      601 cgtccatttt cagtgtcttc atcctgtgca ttgctgccta ccgctctgtc cagcagcccc
      661 tcaggtacct taagtatcgt accaagaccc gagcctcggc caccattctg ggggcctggt
      721 ttctctcttt tctgtgggtt attcccattc taggctggaa tcacttcatg cagcagacct
      781 cggtgcgccg agaggacaag tgtgagacag acttctatga tgtcacctgg ttcaaggtca
      841 tgactgccat catcaacttc tacctgccca ccttgctcat gctctggttc tatgccaaga
      901 tctacaaggc cgtacgacaa cactgccagc accgggagct catcaatagg tccctccctt
      961 ccttctcaga aattaagctg aggccagaga accccaaggg ggatgccaag aaaccaggga
     1021 aggagtctcc ctgggaggtt ctgaaaagga agccaaaaga tgctggtggt ggatctgtct
     1081 tgaagtcacc atcccaaacc cccaaggaga tgaaatcccc agttgtcttc agccaagagg
     1141 atgatagaga agtagacaaa ctctactgct ttccacttga tattgtgcac atgcaggctg
     1201 cggcagaggg gagtagcagg gactatgtag ccgtcaaccg gagccatggc cagctcaaga
     1261 cagatgagca gggcctgaac acacatgggg ccagcgagat atcagaggat cagatgttag
     1321 gtgatagcca gtccttctct cgaacggact cagataccac cacagagaca gcaccaggca
     1381 aaggcaaatt gaggagtggg tctaacacag gcctggatta catcaagttt acttggaaga
     1441 ggctccgctc gcattcaaga cagtatgtat ctgggttgca catgaaccgc gaaaggaagg
     1501 ccgccaaaca gttgggtttt atcatggcag ccttcatcct ctgctggatc ccttatttca
     1561 tcttcttcat ggtcattgcc ttctgcaaga actgttgcaa tgaacatttg cacatgttca
     1621 ccatctggct gggctacatc aactccacac tgaaccccct catctacccc ttgtgcaatg
     1681 agaacttcaa gaagacattc aagagaattc tgcatattcg ctccccgcgg gcccgggatc
     1741 caccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     1801 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     1861 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     1921 tgccctggcc caccctcgtg accaccctgt cctggggcgt gcagtgcttc gcccgctacc
     1981 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     2041 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     2101 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     2161 acatcctggg gcacaagctg gagtacaact actttagcga caacgtctat atcaccgccg
     2221 acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacatc gaggacggcg
     2281 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     2341 tgcccgacaa ccactacctg agcacccagt ccaagctgag caaagacccc aacgagaagc
     2401 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     2461 agctgtacaa gtaaagcggc cgcgactcta gatcataatc agccatacca catttgtaga
     2521 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     2581 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     2641 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     2701 actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
     2761 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     2821 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     2881 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     2941 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     3001 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     3061 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     3121 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     3181 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     3241 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3301 aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     3361 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3421 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3481 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     3541 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     3601 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     3661 cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     3721 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     3781 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     3841 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     3901 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     3961 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     4021 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     4081 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     4141 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     4201 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
     4261 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     4321 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     4381 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     4441 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     4501 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     4561 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     4621 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag
     4681 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     4741 aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     4801 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     4861 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     4921 cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa
     4981 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     5041 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5101 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5161 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5221 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     5281 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5341 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5401 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5461 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5521 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5581 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5641 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5701 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     5761 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcatt agttattaat
     5821 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     5881 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     5941 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     6001 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     6061 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     6121 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atg
//
