LOCUS       sequence-422940-        7018 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  HTR2A receptor for split TEV assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7018)
  AUTHORS   Popovic L, Rossner MJ, Wehr MC
  TITLE     Protocol for identifying properties of ERBB receptor antagonists 
            using the barcoded ERBBprofiler assay.
  JOURNAL   STAR Protoc. 2024 Apr 16;5(2):102987. doi: 
            10.1016/j.xpro.2024.102987.
  PUBMED    38635397
REFERENCE   2  (bases 1 to 7018)
  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: "STAR 
            Protoc."; date: "2024-04-16"; volume: "5(2):102987. doi"; pages: " 
            10.1016/j.xpro.2024.102987"
COMMENT     Sequence Label: pTag4C_SP-HTR2A-V2R-NTEV-tcs-GV-2xHA
FEATURES             Location/Qualifiers
     source          1..7018
                     /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(2076..2100)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2591..2611
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2618..2647
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2657..3097
                     /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             3140..3160
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3311..3544
                     /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      3551..3560
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3563..3589
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    3806..3927
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3843..3862)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3897..3916
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3934..4389)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4071..4092)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4283..4302
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4416..4520
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4522..4879
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4522..4713
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4545..4565)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4730..4865
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4914..5708
                     /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             4914..5708
                     /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(4968..4987)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5578..5597
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5896..5915)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5940..5987
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6316..6904
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6805..6824
                     /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 caggctctac
      661 catggatatt ctttgtgaag aaaatacttc tttgagctca actacgaact ccctaatgca
      721 attaaatgat gacaccaggc tctacagtaa tgactttaac tctggagaag ctaacacttc
      781 tgatgcattt aactggacag tcgactctga aaatcgaacc aacctttcct gtgaagggtg
      841 cctctcaccg tcgtgtctct ccttacttca tctccaggaa aaaaactggt ctgctttact
      901 gacagccgta gtgattattc taactattgc tggaaacata ctcgtcatca tggcagtgtc
      961 cctagagaaa aagctgcaga atgccaccaa ctatttcctg atgtcacttg ccatagctga
     1021 tatgctgctg ggtttccttg tcatgcccgt gtccatgtta accatcctgt atgggtaccg
     1081 gtggcctctg ccgagcaagc tttgtgcagt ctggatttac ctggacgtgc tcttctccac
     1141 ggcctccatc atgcacctct gcgccatctc gctggatcgc tacgtcgcca tccagaatcc
     1201 catccaccac agccgcttca actccagaac taaggcattt ctgaaaatca ttgctgtttg
     1261 gaccatatca gtaggtatat ccatgccaat accagtcttt gggctacagg acgattcgaa
     1321 ggtctttaag gaggggagtt gcttactcgc cgatgataac tttgtcctga tcggctcttt
     1381 tgtgtcattt ttcattccct taaccatcat ggtgatcacc tactttctaa ctatcaagtc
     1441 actccagaaa gaagctactt tgtgtgtaag tgatcttggc acacgggcca aattagcttc
     1501 tttcagcttc ctccctcaga gttctttgtc ttcagaaaag ctcttccagc ggtcgatcca
     1561 tagggagcca gggtcctaca caggcaggag gactatgcag tccatcagca atgagcaaaa
     1621 ggcatgcaag gtgctgggca tcgtcttctt cctgtttgtg gtgatgtggt gccctttctt
     1681 catcacaaac atcatggccg tcatctgcaa agagtcctgc aatgaggatg tcattggggc
     1741 cctgctcaat gtgtttgttt ggatcggtta tctctcttca gcagtcaacc cactagtcta
     1801 cacactgttc aacaagacct ataggtcagc cttttcacgg tatattcagt gtcagtacaa
     1861 ggaaaacaaa aaaccattgc agttaatttt agtgaacaca ataccggctt tggcctacaa
     1921 gtctagccaa cttcaaatgg gacaaaaaaa gaattcaaag caagatgcca agacaacaga
     1981 taatgactgc tcaatggttg ctctaggaaa gcagcattct gaagaggctt ctaaagacaa
     2041 tagcgacgga gtgaatgaaa aggtgagctg tgtggaccca gctttcttgt acaaagtggt
     2101 tgatggtacc gcccggggac gcaccccacc cagcctgggt ccccaagatg agtcctgcac
     2161 caccgccagc tcctccctgg ccaaggacac ttcatcggcc agccctagca accctggcgc
     2221 cagcaacggc agcggagaga gcttgtttaa gggaccacgt gattacaacc cgatatcgag
     2281 caccatttgt catttgacga atgaatctga tgggcacaca acatcgttgt atggtattgg
     2341 atttggtccc ttcatcatta caaacaagca cttgtttcgt cgcaataatg gaacactgtt
     2401 ggtccaatca ctacatggtg tattcaaggt caaggacacc acgactttgc aacaacacct
     2461 cattgatggg agggacatga tgattattcg catgcctaag gatttcccac catttcctca
     2521 aaagctgaaa tttagagagc cacaaaggga agagcgcata tgtcttgtga caaccaactt
     2581 ccaagctccc gagaacctgt acttccaggg ttctagagaa cagaagctga tctcagaaga
     2641 ggatctggcc cgggccatga agctactgtc ttctatcgaa caagcatgcg atatttgccg
     2701 acttaaaaag ctcaagtgct ccaaagaaaa accgaagtgc gccaagtgtc tgaagaacaa
     2761 ctgggagtgt cgctactctc ccaaaaccaa aaggtctccg ctgactaggg cacatctgac
     2821 agaagtggaa tcaaggctag aaagactgga acagctattt ctactgattt ttcctcgaga
     2881 agaccttgac atgattttga aaatggattc tttacaggat ataaaagcat tgttaacagg
     2941 attatttgta caagataatg tgaataaaga tgccgtcaca gatagattgg cttcagtgga
     3001 gactgatatg cctctaacat tgagacagca tagaataagt gcgacatcat catcggaaga
     3061 gagtagtaac aaaggtcaaa gacagttgac tgtatcgccg gaattcccgg ggatcagctt
     3121 gataattcga ttcaccatgc ccaagaagaa gaggaaggtg gcgtacagcc gcgcgcgtac
     3181 gaaaaacaat tacgggtcta ccatcgaggg cctgctcgat ctcccggacg acgacgcccc
     3241 cgaagaggcg gggctggcgg ctccgcgcct gtcctttctc cccgcgggac acacgcgcag
     3301 actgtcgacg gcccccccga ccgatgtcag cctgggggac gagctccact tagacggcga
     3361 ggacgtcgcg atggcgcatg ccgacgcgct agacgatttc gatctggaca tgttggggga
     3421 cggggattcc ccgggtccgg gatttacccc ccacgactcc gccccctacg gcgctctgga
     3481 tatggccgac ttcgagtttg agcagatgtt taccgatgcc cttggaattg acgagtacgg
     3541 tggggcggcc gccaccatgg ggtatccata tgatgttcca gattatgcta gatctggaat
     3601 cactagtgaa ttatcgaatt ctgcagccaa gctaattccg ggcgaatgtc ttatgattta
     3661 tgatttttat tattaacctg cagggcggcc gcgactctag atcataatca gccataccac
     3721 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     3781 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     3841 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     3901 tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat attttgttaa
     3961 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     4021 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     4081 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     4141 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     4201 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     4261 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     4321 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     4381 agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     4441 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     4501 aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     4561 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4621 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4681 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     4741 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     4801 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     4861 aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg
     4921 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     4981 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     5041 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg
     5101 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     5161 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     5221 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     5281 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     5341 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     5401 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg
     5461 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     5521 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     5581 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     5641 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     5701 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     5761 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     5821 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     5881 tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     5941 ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt
     6001 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     6061 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     6121 agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat
     6181 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     6241 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     6301 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     6361 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     6421 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     6481 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6541 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6601 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6661 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6721 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6781 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6841 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6901 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6961 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcat
//
