LOCUS       sequence-423236-        7428 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  barcoded assay, MAPK sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7428)
  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 7428)
  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: pGL4_EGR1p-BC1489-luc2
FEATURES             Location/Qualifiers
     source          1..7428
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    47..71
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(117..138)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    complement(272..292)
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    1249..1269
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        1296..1314
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(1378..1396)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    complement(1420..1444)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1502..1511
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1508..3160
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     CDS             3164..3211
                     /codon_start=1
                     /product="non-ORF yeast peptide conferring 
                     ubiquitin-dependent degradation"
                     /label=hCL1
                     /note="human codon-optimized"
                     /translation="ACKNWFSSLSHFVIHL"
     CDS             3215..3334
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    3383..3504
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3394..3413)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3448..3467
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3698..4055
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3698..3889
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3721..3741)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3906..4041
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3968..3987
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4086..5123
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             5102..5110
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    5147..5195
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5351..5368)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5522..6110)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5602..5621)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6310..7101,7102..7170))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    7275..7323
                     /note="synthetic polyadenylation signal"
     misc_feature    7337..7428
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     7377..7396
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagctcgcta gcctcgagga tatcaaacaa gtttgtacaa
       61 aaaagcaggc tatggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
      121 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
      181 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
      241 cctggcccac cctcgtgacc accctgacct acaacttttc tatacaaagt tgagacaccg
      301 tgccatagat cgaggcccgg ggtcaaggcc ccgcctctcc tgggcggccc ctgcccaggc
      361 gggcccagcc gctcctcccc cgcactcccg gttcgctctc acggtccctg aggtgggcgg
      421 gcgggccctg gatgacagcg atagaacccc ggcccgactc gccctcgccc ccgctctggg
      481 tctgggcttc cccagcctag ttcacgccta ggagccgcct gagcagccgc gcgcccagcg
      541 ccacacgcca cgagccctcc ccgcctgggc gtccccggat cccgcgagcg ctcgggctcc
      601 cggcttggaa ccagggagga gggagggagc gagggagcaa ccagctgcga cccggaaatg
      661 ccatataagg agcaggaagg atcccccgcc ggaacaaccc ttatttgggc agcaccttat
      721 ttggagtggc ccgatatggc ccggccgctt ccggctctgg gaggagggaa gaaggcggag
      781 ggaggggcaa cgcgggaact ccggagctgc gcgggtcccg gaggccccgg cggcggctag
      841 agctctaggc ttccccgaag cctgggcgcc tgggatgcgg gcgcgggcgc gggccctagg
      901 gtgcaggatg gaggtgccgg gcgctgtcgg atggggggct tcacgtcgct ccgggtcctc
      961 ccggccggtc ctgccatatt agggcttcct gcttcccata tatggccatg tacgtcacga
     1021 cggaggcgga cccgtgccgt tccagaccct tcaaatagag gcggatccgg ggagtcgcga
     1081 gagatcccag cgcgcagaac ttggggagcc gccgccgcca tccgccgccg cagccagctt
     1141 ccgccgccgc aggaccggcc cctgccccag cctccgcagc cgcggcgcgt ccacgcccgc
     1201 ccgcgcccag ggcgagtcgg ggtcgccgcc tgcacgcttc tcagtgttca actttgtata
     1261 ataaagttga gctagttgct aagtctgccg agtagaatta accctcacta aagggtaggt
     1321 gacactatac atctttaatc caaactttac accgagactt tcttttactt tactactcct
     1381 atagtgagtc gtattagtag acgcgagtca atgcatgtac gacagctttc ttgtacaaag
     1441 tggttcgata tcaagatctg gcctcggcgg ccaagcttgg caatccggta ctgttggtaa
     1501 agccaccatg gaagatgcca aaaacattaa gaagggccca gcgccattct acccactcga
     1561 agacgggacc gccggcgagc agctgcacaa agccatgaag cgctacgccc tggtgcccgg
     1621 caccatcgcc tttaccgacg cacatatcga ggtggacatt acctacgccg agtacttcga
     1681 gatgagcgtt cggctggcag aagctatgaa gcgctatggg ctgaatacaa accatcggat
     1741 cgtggtgtgc agcgagaata gcttgcagtt cttcatgccc gtgttgggtg ccctgttcat
     1801 cggtgtggct gtggccccag ctaacgacat ctacaacgag cgcgagctgc tgaacagcat
     1861 gggcatcagc cagcccaccg tcgtattcgt gagcaagaaa gggctgcaaa agatcctcaa
     1921 cgtgcaaaag aagctaccga tcatacaaaa gatcatcatc atggatagca agaccgacta
     1981 ccagggcttc caaagcatgt acaccttcgt gacttcccat ttgccacccg gcttcaacga
     2041 gtacgacttc gtgcccgaga gcttcgaccg ggacaaaacc atcgccctga tcatgaacag
     2101 tagtggcagt accggattgc ccaagggcgt agccctaccg caccgcaccg cttgtgtccg
     2161 attcagtcat gcccgcgacc ccatcttcgg caaccagatc atccccgaca ccgctatcct
     2221 cagcgtggtg ccatttcacc acggcttcgg catgttcacc acgctgggct acttgatctg
     2281 cggctttcgg gtcgtgctca tgtaccgctt cgaggaggag ctattcttgc gcagcttgca
     2341 agactataag attcaatctg ccctgctggt gcccacacta tttagcttct tcgctaagag
     2401 cactctcatc gacaagtacg acctaagcaa cttgcacgag atcgccagcg gcggggcgcc
     2461 gctcagcaag gaggtaggtg aggccgtggc caaacgcttc cacctaccag gcatccgcca
     2521 gggctacggc ctgacagaaa caaccagcgc cattctgatc acccccgaag gggacgacaa
     2581 gcctggcgca gtaggcaagg tggtgccctt cttcgaggct aaggtggtgg acttggacac
     2641 cggtaagaca ctgggtgtga accagcgcgg cgagctgtgc gtccgtggcc ccatgatcat
     2701 gagcggctac gttaacaacc ccgaggctac aaacgctctc atcgacaagg acggctggct
     2761 gcacagcggc gacatcgcct actgggacga ggacgagcac ttcttcatcg tggaccggct
     2821 gaagagcctg atcaaataca agggctacca ggtagcccca gccgaactgg agagcatcct
     2881 gctgcaacac cccaacatct tcgacgccgg ggtcgccggc ctgcccgacg acgatgccgg
     2941 cgagctgccc gccgcagtcg tcgtgctgga acacggtaaa accatgaccg agaaggagat
     3001 cgtggactat gtggccagcc aggttacaac cgccaagaag ctgcgcggtg gtgttgtgtt
     3061 cgtggacgag gtgcctaaag gactgaccgg caagttggac gcccgcaaga tccgcgagat
     3121 tctcattaag gccaagaagg gcggcaagat cgccgtgtaa tctgcttgca agaactggtt
     3181 cagtagctta agccactttg tgatccacct taacagccac ggcttccctc ccgaggtgga
     3241 ggagcaggcc gccggcaccc tgcccatgag ctgcgcccag gagagcggca tggatagaca
     3301 ccctgctgct tgcgccagcg ccaggatcaa cgtctaactg cagtctagag tcggggcggc
     3361 cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga
     3421 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     3481 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     3541 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc
     3601 gataaggatc cgtttgcgta ttgggcgctc ttccgctgat ctgcgcagca ccatggcctg
     3661 aaataacctc tgaaagagga acttggttag ctaccttctg aggcggaaag aaccagctgt
     3721 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3781 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3841 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3901 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3961 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     4021 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cgattcttct gacactagcg
     4081 ccaccatgaa gaagcccgaa ctcaccgcta ccagcgttga aaaatttctc atcgagaagt
     4141 tcgacagtgt gagcgacctg atgcagttgt cggagggcga agagagccga gccttcagct
     4201 tcgatgtcgg cggacgcggc tatgtactgc gggtgaatag ctgcgctgat ggcttctaca
     4261 aagaccgcta cgtgtaccgc cacttcgcca gcgctgcact acccatcccc gaagtgttgg
     4321 acatcggcga gttcagcgag agcctgacat actgcatcag tagacgcgcc caaggcgtta
     4381 ctctccaaga cctccccgaa acagagctgc ctgctgtgtt acagcctgtc gccgaagcta
     4441 tggatgctat tgccgccgcc gacctcagtc aaaccagcgg cttcggccca ttcgggcccc
     4501 aaggcatcgg ccagtacaca acctggcggg atttcatttg cgccattgct gatccccatg
     4561 tctaccactg gcagaccgtg atggacgaca ccgtgtccgc cagcgtagct caagccctgg
     4621 acgaactgat gctgtgggcc gaagactgtc ccgaggtgcg ccacctcgtc catgccgact
     4681 tcggcagcaa caacgtcctg accgacaacg gccgcatcac cgccgtaatc gactggtccg
     4741 aagctatgtt cggggacagt cagtacgagg tggccaacat cttcttctgg cggccctggc
     4801 tggcttgcat ggagcagcag actcgctact tcgagcgccg gcatcccgag ctggccggca
     4861 gccctcgtct gcgagcctac atgctgcgca tcggcctgga tcagctctac cagagcctcg
     4921 tggacggcaa cttcgacgat gctgcctggg ctcaaggccg ctgcgatgcc atcgtccgca
     4981 gcggggccgg caccgtcggt cgcacacaaa tcgctcgccg gagcgcagcc gtatggaccg
     5041 acggctgcgt cgaggtgctg gccgacagcg gcaaccgccg gcccagtaca cgaccgcgcg
     5101 ctaaggaggt aggtcgagtt taaactctag aaccggtcat ggccgcaata aaatatcttt
     5161 attttcatta catctgtgtg ttggtttttt gtgtgttcga actagatgct gtcgaccgat
     5221 gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt
     5281 cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct
     5341 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     5401 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     5461 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     5521 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     5581 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     5641 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     5701 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     5761 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     5821 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     5881 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     5941 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     6001 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     6061 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     6121 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     6181 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     6241 aatcaatcta aagtatatat gagtaaactt ggtctgacag cggccgcaaa tgctaaacca
     6301 ctgcagtggt taccagtgct tgatcagtga ggcaccgatc tcagcgatct gcctatttcg
     6361 ttcgtccata gtggcctgac tccccgtcgt gtagatcact acgattcgtg agggcttacc
     6421 atcaggcccc agcgcagcaa tgatgccgcg agagccgcgt tcaccggccc ccgatttgtc
     6481 agcaatgaac cagccagcag ggagggccga gcgaagaagt ggtcctgcta ctttgtccgc
     6541 ctccatccag tctatgagct gctgtcgtga tgctagagta agaagttcgc cagtgagtag
     6601 tttccgaaga gttgtggcca ttgctactgg catcgtggta tcacgctcgt cgttcggtat
     6661 ggcttcgttc aactctggtt cccagcggtc aagccgggtc acatgatcac ccatattatg
     6721 aagaaatgca gtcagctcct tagggcctcc gatcgttgtc agaagtaagt tggccgcggt
     6781 gttgtcgctc atggtaatgg cagcactaca caattctctt accgtcatgc catccgtaag
     6841 atgcttttcc gtgaccggcg agtactcaac caagtcgttt tgtgagtagt gtatacggcg
     6901 accaagctgc tcttgcccgg cgtctatacg ggacaacacc gcgccacata gcagtacttt
     6961 gaaagtgctc atcatcggga atcgttcttc ggggcggaaa gactcaagga tcttgccgct
     7021 attgagatcc agttcgatat agcccactct tgcacccagt tgatcttcag catcttttac
     7081 tttcaccagc gtttcggggt gtgcaaaaac aggcaagcaa aatgccgcaa agaagggaat
     7141 gagtgcgaca cgaaaatgtt ggatgctcat actcgtcctt tttcaatatt attgaagcat
     7201 ttatcagggt tactagtacg tctctcaagg ataagtaagt aatattaagg tacgggaggt
     7261 attggacagg ccgcaataaa atatctttat tttcattaca tctgtgtgtt ggttttttgt
     7321 gtgaatcgat agtactaaca tacgctctcc atcaaaacaa aacgaaacaa aacaaactag
     7381 caaaataggc tgtccccagt gcaagtgcag gtgccagaac atttctct
//
