LOCUS       sequence-456026-        6928 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  DRD2 receptor for split TEV assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6928)
  AUTHORS   Popovic L, Brankatschk B, Palladino G, Rossner MJ, Wehr MC
  TITLE     Polypharmacological profiling across protein target families and 
            cellular pathways using the multiplexed cell-based assay platform 
            safetyProfiler reveals efficacy, potency and side effects of drugs.
  JOURNAL   Biomed Pharmacother. 2024 Nov;180:117523. doi: 
            10.1016/j.biopha.2024.117523. Epub 2024 Oct 14.
  PUBMED    39405910
REFERENCE   2  (bases 1 to 6928)
  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"; journalName: "Biomed 
            Pharmacother."; date: "2024-11"; volume: "180:117523. doi"; pages: "
            10.1016/j.biopha.2024.117523. Epub 2024 Oct 14"
COMMENT     Sequence Label: pTag4C_SP-DRD2-V2R-NTEV-tcs-GV-2xHA
FEATURES             Location/Qualifiers
     source          1..6928
                     /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"
     protein_bind    632..656
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1986..2010)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2501..2521
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2528..2557
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2567..3007
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     CDS             3050..3070
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3221..3454
                     /codon_start=1
                     /gene="UL48"
                     /product="transcriptional activation domain of herpes 
                     simplex virus protein VP16 (Triezenberg et al., 1988; 
                     Cousens et al., 1989)"
                     /label=VP16 AD
                     /translation="APPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGF
                     TPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     regulatory      3461..3470
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3473..3499
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    3716..3837
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3753..3772)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3807..3826
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3844..4299)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3981..4002)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4193..4212
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4326..4430
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4432..4789
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4432..4623
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4455..4475)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4640..4775
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4824..5618
                     /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             4824..5618
                     /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(4878..4897)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5488..5507
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5806..5825)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5850..5897
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6226..6814
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6715..6734
                     /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 caagcttatc aacaagtttg tacaaaaaag caggctccac
      661 catgaagacg atcatcgccc tgagctacat cttctgcctg gtattcgccg actacaagga
      721 cgatgatgac gccagcatcg atatggatcc actgaatctg tcctggtatg atgatgatct
      781 ggagaggcag aactggagcc ggcccttcaa cgggtcagac gggaaggcgg acagacccca
      841 ctacaactac tatgccacac tgctcaccct gctcatcgct gtcatcgtct tcggcaacgt
      901 gctggtgtgc atggctgtgt ctagagagaa ggcgctgcag accaccacca actacctgat
      961 cgtcagcctc gcagtggccg acctcctcgt cgccacactg gtcatgccct gggttgtcta
     1021 cctggaggtg gtaggtgagt ggaaattcag caggattcac tgtgacatct tcgtcactct
     1081 ggacgtcatg atgtgcacgg cgagcatcct gaacttgtgt gccatcagca tcgacaggta
     1141 cacagctgtg gccatgccca tgctgtacaa tacgcgctac agctccaagc gccgggtcac
     1201 cgtcatgatc tccatcgtct gggtcctgtc cttcaccatc tcctgcccac tcctcttcgg
     1261 actcaataac gcagaccaga acgagtgcat cattgccaac ccggccttcg tggtctactc
     1321 ctccatcgtc tccttctacg tgcccttcat tgtcaccctg ctggtctaca tcaagatcta
     1381 cattgtcctc cgcagacgcc gcaagcgagt caacaccaaa cgcagcagcc gagctttcag
     1441 ggcccacctg agggctccac taaaggaggc tgcccggcga gcccaggagc tggagatgga
     1501 gatgctctcc agcaccagcc cacccgagag gacccggtac agccccatcc cacccagcca
     1561 ccaccagctg actctccccg acccgtccca ccacggtctc cacagcactc ctgacagccc
     1621 cgccaaacca gagaagaatg ggcatgccaa agaccacccc aagattgcca agatctttga
     1681 gatccagacc atgcccaatg gcaaaacccg gacctccctc aagaccatga gccgtaggaa
     1741 gctctcccag cagaaggaga agaaagccac tcagatgctc gccattgttc tcggcgtgtt
     1801 catcatctgc tggctgccct tcttcatcac acacatcctg aacatacact gtgactgcaa
     1861 catcccgcct gtcctgtaca gcgccttcac gtggctgggc tatgtcaaca gcgccgtgaa
     1921 ccccatcatc tacaccacct tcaacattga gttccgcaag gccttcctga agatcctcca
     1981 ctgcgaccca gctttcttgt acaaagtggt tgatggtacc gcccggggac gcaccccacc
     2041 cagcctgggt ccccaagatg agtcctgcac caccgccagc tcctccctgg ccaaggacac
     2101 ttcatcggcc agccctagca accctggcgc cagcaacggc agcggagaga gcttgtttaa
     2161 gggaccacgt gattacaacc cgatatcgag caccatttgt catttgacga atgaatctga
     2221 tgggcacaca acatcgttgt atggtattgg atttggtccc ttcatcatta caaacaagca
     2281 cttgtttcgt cgcaataatg gaacactgtt ggtccaatca ctacatggtg tattcaaggt
     2341 caaggacacc acgactttgc aacaacacct cattgatggg agggacatga tgattattcg
     2401 catgcctaag gatttcccac catttcctca aaagctgaaa tttagagagc cacaaaggga
     2461 agagcgcata tgtcttgtga caaccaactt ccaagctccc gagaacctgt acttccaggg
     2521 ttctagagaa cagaagctga tctcagaaga ggatctggcc cgggccatga agctactgtc
     2581 ttctatcgaa caagcatgcg atatttgccg acttaaaaag ctcaagtgct ccaaagaaaa
     2641 accgaagtgc gccaagtgtc tgaagaacaa ctgggagtgt cgctactctc ccaaaaccaa
     2701 aaggtctccg ctgactaggg cacatctgac agaagtggaa tcaaggctag aaagactgga
     2761 acagctattt ctactgattt ttcctcgaga agaccttgac atgattttga aaatggattc
     2821 tttacaggat ataaaagcat tgttaacagg attatttgta caagataatg tgaataaaga
     2881 tgccgtcaca gatagattgg cttcagtgga gactgatatg cctctaacat tgagacagca
     2941 tagaataagt gcgacatcat catcggaaga gagtagtaac aaaggtcaaa gacagttgac
     3001 tgtatcgccg gaattcccgg ggatcagctt gataattcga ttcaccatgc ccaagaagaa
     3061 gaggaaggtg gcgtacagcc gcgcgcgtac gaaaaacaat tacgggtcta ccatcgaggg
     3121 cctgctcgat ctcccggacg acgacgcccc cgaagaggcg gggctggcgg ctccgcgcct
     3181 gtcctttctc cccgcgggac acacgcgcag actgtcgacg gcccccccga ccgatgtcag
     3241 cctgggggac gagctccact tagacggcga ggacgtcgcg atggcgcatg ccgacgcgct
     3301 agacgatttc gatctggaca tgttggggga cggggattcc ccgggtccgg gatttacccc
     3361 ccacgactcc gccccctacg gcgctctgga tatggccgac ttcgagtttg agcagatgtt
     3421 taccgatgcc cttggaattg acgagtacgg tggggcggcc gccaccatgg ggtatccata
     3481 tgatgttcca gattatgcta gatctggaat cactagtgaa ttatcgaatt ctgcagccaa
     3541 gctaattccg ggcgaatgtc ttatgattta tgatttttat tattaacctg cagggcggcc
     3601 gcgactctag atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     3661 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     3721 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     3781 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg
     3841 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     3901 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     3961 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     4021 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     4081 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     4141 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     4201 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     4261 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     4321 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     4381 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg
     4441 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     4501 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     4561 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     4621 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     4681 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     4741 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag
     4801 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     4861 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     4921 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     4981 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
     5041 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     5101 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     5161 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     5221 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     5281 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     5341 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     5401 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     5461 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     5521 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     5581 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     5641 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     5701 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     5761 ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga
     5821 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     5881 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     5941 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     6001 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     6061 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     6121 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     6181 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     6241 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     6301 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     6361 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     6421 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     6481 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     6541 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     6601 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     6661 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6721 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     6781 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     6841 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     6901 tgtggataac cgtattaccg ccatgcat
//
