LOCUS       sequence-198412-        8539 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of EPHA3 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8539)
  AUTHORS   Zhuang G, Song W, Amato K, Hwang Y, Lee K, Boothby M, Ye F, Guo Y, 
            Shyr Y, Lin L, Carbone DP, Brantley-Sieders DM, Chen J
  TITLE     Effects of cancer-associated EPHA3 mutations on lung cancer.
  JOURNAL   J Natl Cancer Inst. 2012 Aug 8;104(15):1182-97. doi: 
            10.1093/jnci/djs297. Epub 2012 Jul 24.
  PUBMED    22829656
REFERENCE   2  (bases 1 to 8539)
  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: 
            "10.1093/jnci/djs297"; journalName: "J Natl Cancer Inst"; date: 
            "2012-08-8- 8"; volume: "104"; issue: "15"; pages: "1182-97"
COMMENT     Sequence Label: T166N EPHA3 pcDNA3.1
FEATURES             Location/Qualifiers
     source          1..8539
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     148..168
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(177..605)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(287..308)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     499..518
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    651..875
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     864..881
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(904..921)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(937..966)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(4127..4146)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4128..4146)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4191..4394)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4220..4240)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4395..4774
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     4946..4965
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5040..5144
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5145..5213,5214..6005)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5363..5382)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6176..6764
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6665..6684
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6918..6935
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7052..7073
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7088..7093,7094..7111,7112..7118)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7126..7142
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     7131..7153
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7150..7166
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7150..7166
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    7215..7336
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7226..7245)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7280..7299
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(7512..8306)
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(7623..7642)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     8233..8252
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(8373..8539,1..163))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8387..8522
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8441..8460)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 ggactatggt tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc
       61 ctggggactt tccacacctg gttgctgact aattgagatg catgctttgc atacttctgc
      121 ctgctgggga gcctggggac tttccacacc ctaactgaca cacattccac agaattaatt
      181 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
      241 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
      301 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
      361 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
      421 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
      481 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
      541 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
      601 cgcgtgggga taccccctag agccccagct ggttctttcc gcctcagaag ccatagagcc
      661 caccgcatcc ccagcatgcc tgctattgtc ttcccaatcc tcccccttgc tgtcctgccc
      721 caccccaccc cccagaatag aatgacacct actcagacaa tgcgatgcaa tttcctcatt
      781 ttattaggaa aggacagtgg gagtggcacc ttccagggtc aaggaaggca cgggggaggg
      841 gcaaacaaca gatggctggc aactagaagg cacagtcgag gctgatcagc gggtttaaac
      901 tcaatggtga tggtgatgat gaccggtatg catattcaga tcctcttctg agatgagttt
      961 ttgttcgaac cgcgggccct ctagagcttt aatgctactg atgatcttct tctgtggccc
     1021 aaccacggtg acaccaacct ttttcatgtc atctgtggaa atcttggcta ttgtgtcaca
     1081 agaactgtac tccacacccg tgaagatttc cttgcagtgt gctgtccaga caccattaag
     1141 ccagtcacct gttgtgcgga aggtagtgat atccacattg ctttggtcca gaagaaggtt
     1201 tgatggcctt gcggctgcac tggtgatgat cttcaggctg ccgggattcc ggataagctt
     1261 gtccagaata ctaacaatct gctcaaactt gggtctgttg ttcctgtctt tctgccagca
     1321 gtccagcatc agctgataca aggcagctgg gcagtccatg gggggtggca gtcgatagcc
     1381 ctcatctaca gctttaatta catcctgatt ggacatctcc cagtatggtc tctctccata
     1441 agacatcacc tcccagagaa caatcccata actccataca tcgctggctg acgtgaactt
     1501 gcggtaggct atagcttctg gtgatgtcca cctgattggg atcttccctc ctcttgttgt
     1561 ataagcagct tctgggtcat cctccaggac acgcgaaagt ccgaaatcag aaaccttaca
     1621 caccaagtta ctgttgatca agatgttccg agcagcgagg tctcggtgaa catagcccat
     1681 gtctgacagg tacttcatgc cagatgctat ccctcgaagc atccccacta gctgaatgac
     1741 agtaaactgg gcatcgtgtt tacgtaggaa actatccaag gaaccattct ccatgtattc
     1801 tgtgacaatc ataactggct tacttttggt aacaactcct tccagtcgaa tgatattggg
     1861 gtggtcaaac tgtcccataa tgcttgcttc tcccaggaag tctctcctct gcttttctgt
     1921 gtagccaact ttcagggtct taatggccac tgaaatctct ttttttgaag gaagttttaa
     1981 gcgaccactg cacacctctc caaattcacc tgctccaaca actttatcaa tggatatgtt
     2041 ggtggcatcc aattccttgg caaactcatg aacagcttgg gtagggtctt catatgtatg
     2101 tgggtcaaca taagtcctga gacctggaag ttttaaatgc ccattgccaa aatgaagtct
     2161 tttttcatct gccccatgtt ttgacttata gccacagaac ctcccaatca aaacatagat
     2221 gacaacagtg aggagaataa ttgctactgc cgctgaaatg gcgatcatga ccacttggct
     2281 actttcacca gagatggaga aagagtctgg actagtttca aactcaaact tgcggctgtt
     2341 cgtcccatat ccagcggctg ttcgggctcg gatttggaat acgtatatag tgtcaggctt
     2401 gaggctactg atggtaacat ttgtgcctct tgccctcaga atggtataac ttgtttcttg
     2461 ttcctgcttt tcatagtatt tgacctcgta gtccaatatg atcccattag gatgttcagg
     2521 ttcttgccag gacaaagaga tgctatttct ggaggtccga tctttcttaa tcgtcaggac
     2581 aggtgatgga gcagcctgat tagttgtgat gctgaccgca gcaaactgtc ttggtgggga
     2641 gctcagctct gacaccccat taacggcatc aatctcaaag gtgtagttag tatgtgccag
     2701 aaggtctgtc actgtcaccg tggtgttggt gagtccaaac tgtcgaggga ggaagcggac
     2761 atttgggctg catggctcac actgttttat attccaccca cattttttac atatgatgtt
     2821 gaaggtaaca tctttccggc ctcctgtgtc caggggccaa ctccagtcca ggataactga
     2881 ggtctcgttt atattagaga taacatttct tggtgaagat ggaggtcggg tacaagccat
     2941 ggatggaggg tctttgtctg cccggaagta attattctca cacctgcagt tcattgaacc
     3001 atcttcctga gtagaactgt gaggcgggca cttagcacac ttcatattac catccaatgc
     3061 cttgtagaaa cctggtcgac aagcttggca cataaaacct ctttcttcat agccagcatt
     3121 gcaggaacac ttgccaatgg gtacaagcca ttcgccttct gtactgcagt acatccttgg
     3181 aggatcttcc tccttagaat tgttgacaca agaccctcta acctccacca gggactggga
     3241 gtccatgggt accgtgtctg gaaacatagc cagattcttc actgtaaatg ggcacttttt
     3301 gaagtatact ctcacagaca ccaaggcaac acaagcacca acatcttgaa atgccaaata
     3361 aaatcccttc ttgttgacag gacctacttc tctaatctca ttgttgagct tcagaatacg
     3421 gtccccaaga tccatttgag tgaaactttc atcagctgca atggtgtcaa tctttgtaaa
     3481 ctgatgctct cgaaatttca ccccatgatc atcatcagac tccatgtagt acaggttgaa
     3541 tgtctccttg caagttccta aaaccaatgg aatgctattg cagtctcgta gagtgaactt
     3601 gagctccaca taaatcttct gagctgagtt cctggggacc cagtttgttc tcagccaatt
     3661 gttttgactg tggtccatga cattgcacac ctggtaagtc ctgatgggtg tgtaatgttc
     3721 atccacacca ctgatctctt cccacccatg tgatggataa gagatccagc ccagctcccc
     3781 ttgaattgtt tttgaatcca gtagattgac ttcattggaa ggctgcggaa tcagttcccc
     3841 gaagctgtcg agaacagagc agctgagaag gaggaggatg gagagctgac aatccatgtt
     3901 gctggtgcag agggcagtga gaggagcata tctccatgaa gcatgccact gatgtgaggg
     3961 ggagcgctct gattgctgga gaagttacca tgctccgctc gcaggctgat gtcaagtttg
     4021 acactcctgg ttctacgctg tcacctcgct agagctcgac gaattccacc acactggact
     4081 agtggatccg agctcggtac caagcttaac tagccagctt gggtctccct atagtgagtc
     4141 gtattaattt cgataagcca gtaagcagtg ggttctctag ttagccagag agctctgctt
     4201 atatagacct cccaccgtac acgcctaccg cccatttgcg tcaatggggc ggagttgtta
     4261 cgacattttg gaaagtcccg ttgattttgg tgccaaaaca aactcccatt gacgtcaatg
     4321 gggtggagac ttggaaatcc ccgtgagtca aaccgctatc cacgcccatt gatgtactgc
     4381 caaaaccgca tcaccatggt aatagcgatg actaatacgt agatgtactg ccaagtagga
     4441 aagtcccata aggtcatgta ctgggcataa tgccaggcgg gccatttacc gtcattgacg
     4501 tcaatagggg gcgtacttgg catatgatac acttgatgta ctgccaagtg ggcagtttac
     4561 cgtaaatact ccacccattg acgtcaatgg aaagtcccta ttggcgttac tatgggaaca
     4621 tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg cgggccattt
     4681 accgtaagtt atgtaacgcg gaactccata tatgggctat gaactaatga ccccgtaatt
     4741 gattactatt aataactagt caataatcaa tgtcaacgcg tatatctggc ccgtacatcg
     4801 cgaagcagcg caaaacgcct aaccctaagc agattcttca tgcaattgtc ggtcaagcct
     4861 tgccttgttg tagcttaaat tttgctcgcg cactactcag cgacctccaa cacacaagca
     4921 gggagcagat actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     4981 aggggatcgg gagatctccc gatccgtcga cgtcaggtgg cacttttcgg ggaaatgtgc
     5041 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     5101 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     5161 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     5221 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     5281 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     5341 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     5401 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     5461 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     5521 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     5581 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     5641 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     5701 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     5761 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     5821 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     5881 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     5941 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     6001 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     6061 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     6121 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     6181 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     6241 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     6301 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     6361 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     6421 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     6481 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     6541 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     6601 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     6661 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     6721 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     6781 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     6841 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     6901 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     6961 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     7021 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
     7081 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     7141 aatttcacac aggaaacagc tatgaccatg attacgccaa gctctagcta gaggtcgacg
     7201 gtatacagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     7261 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     7321 ctgcaataaa caagttgggg tgggcgaaga actccagcat gagatccccg cgctggagga
     7381 tcatccagcc ggcgtcccgg aaaacgattc cgaagcccaa cctttcatag aaggcggcgg
     7441 tggaatcgaa atctcgtgat ggcaggttgg gcgtcgcttg gtcggtcatt tcgcgaaccc
     7501 cagagtcccg ctcagaagaa ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg
     7561 ggagcggcga taccgtaaag cacgaggaag cggtcagccc attcgccgcc aagctcttca
     7621 gcaatatcac gggtagccaa cgctatgtcc tgatagcggt ccgccacacc cagccggcca
     7681 cagtcgatga atccagaaaa gcggccattt tccaccatga tattcggcaa gcaggcatcg
     7741 ccatgggtca cgacgagatc ctcgccgtcg ggcatgcgcg ccttgagcct ggcgaacagt
     7801 tcggctggcg cgagcccctg atgctcttcg tccagatcat cctgatcgac aagaccggct
     7861 tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta
     7921 gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca tgatggatac tttctcggca
     7981 ggagcaaggt gagatgacag gagatcctgc cccggcactt cgcccaatag cagccagtcc
     8041 cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc
     8101 cacgatagcc gcgctgcctc gtcctgcagt tcattcaggg caccggacag gtcggtcttg
     8161 acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca cggcggcatc agagcagccg
     8221 attgtctgtt gtgcccagtc atagccgaat agcctctcca cccaagcggc cggagaacct
     8281 gcgtgcaatc catcttgttc aatcatgcga aacgatcctc atcctgtctc ttgatcagat
     8341 ccgaaaatgg atatacaagc tcccgggagc tttttgcaaa agcctaggcc tccaaaaaag
     8401 cctcctcact acttctggaa tagctcagag gcagaggcgg cctcggcctc tgcataaata
     8461 aaaaaaatta gtcagccatg gggcggagaa tgggcggaac tgggcggagt taggggcggg
     8521 atgggcggag ttaggggcg
//
