LOCUS       sequence-194260-        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: EPHA3_1-972 pcDNA3.1
FEATURES             Location/Qualifiers
     source          1..8539
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4043..4072
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4088..4105
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4128..4145)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4134..4358
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4404..4832
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4491..4510)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4701..4722
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4841..4861)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4846..5175
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5026..5161
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5088..5107
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5242..6036
                     /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(5296..5315)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5906..5925
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6212..6333
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6249..6268)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6303..6322
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6382..6398)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6382..6398)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6395..6417)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6406..6422
                     /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)."
     promoter        complement(join(6430..6436,6437..6454,6455..6460))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6475..6496
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6613..6630)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6784..7372)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6864..6883)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7543..8334,8335..8403))
                     /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     8166..8185
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8404..8508)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 taagcttggt accgagctcg gatccactag tccagtgtgg tggaattcgt cgagctctag
      961 cgaggtgaca gcgtagaacc aggagtgtca aacttgacat cagcctgcga gcggagcatg
     1021 gtaacttctc cagcaatcag agcgctcccc ctcacatcag tggcatgctt catggagata
     1081 tgctcctctc actgccctct gcaccagcaa catggattgt cagctctcca tcctcctcct
     1141 tctcagctgc tctgttctcg acagcttcgg ggaactgatt ccgcagcctt ccaatgaagt
     1201 caatctactg gattcaaaaa caattcaagg ggagctgggc tggatctctt atccatcaca
     1261 tgggtgggaa gagatcagtg gtgtggatga acattacaca cccatcagga cttaccaggt
     1321 gtgcaatgtc atggaccaca gtcaaaacaa ttggctgaga acaaactggg tccccaggaa
     1381 ctcagctcag aagatttatg tggagctcaa gttcactcta cgagactgca atagcattcc
     1441 attggtttta ggaacttgca aggagacatt caacctgtac tacatggagt ctgatgatga
     1501 tcatggggtg aaatttcgag agcatcagtt tacaaagatt gacaccattg cagctgatga
     1561 aagtttcact caaatggatc ttggggaccg tattctgaag ctcaacactg agattagaga
     1621 agtaggtcct gtcaacaaga agggatttta tttggcattt caagatgttg gtgcttgtgt
     1681 tgccttggtg tctgtgagag tatacttcaa aaagtgccca tttacagtga agaatctggc
     1741 tatgtttcca gacacggtac ccatggactc ccagtccctg gtggaggtta gagggtcttg
     1801 tgtcaacaat tctaaggagg aagatcctcc aaggatgtac tgcagtacag aaggcgaatg
     1861 gcttgtaccc attggcaagt gttcctgcaa tgctggctat gaagaaagag gttttatgtg
     1921 ccaagcttgt cgaccaggtt tctacaaggc attggatggt aatatgaagt gtgctaagtg
     1981 cccgcctcac agttctactc aggaagatgg ttcaatgaac tgcaggtgtg agaataatta
     2041 cttccgggca gacaaagacc ctccatccat ggcttgtacc cgacctccat cttcaccaag
     2101 aaatgttatc tctaatataa acgagacctc agttatcctg gactggagtt ggcccctgga
     2161 cacaggaggc cggaaagatg ttaccttcaa catcatatgt aaaaaatgtg ggtggaatat
     2221 aaaacagtgt gagccatgca gcccaaatgt ccgcttcctc cctcgacagt ttggactcac
     2281 caacaccacg gtgacagtga cagaccttct ggcacatact aactacacct ttgagattga
     2341 tgccgttaat ggggtgtcag agctgagctc cccaccaaga cagtttgctg cggtcagcat
     2401 cacaactaat caggctgctc catcacctgt cctgacgatt aagaaagatc ggacctccag
     2461 aaatagcatc tctttgtcct ggcaagaacc tgaacatcct aatgggatca tattggacta
     2521 cgaggtcaaa tactatgaaa agcaggaaca agaaacaagt tataccattc tgagggcaag
     2581 aggcacaaat gttaccatca gtagcctcaa gcctgacact atatacgtat tccaaatccg
     2641 agcccgaaca gccgctggat atgggacgaa cagccgcaag tttgagtttg aaactagtcc
     2701 agactctttc tccatctctg gtgaaagtag ccaagtggtc atgatcgcca tttcagcggc
     2761 agtagcaatt attctcctca ctgttgtcat ctatgttttg attgggaggt tctgtggcta
     2821 taagtcaaaa catggggcag atgaaaaaag acttcatttt ggcaatgggc atttaaaact
     2881 tccaggtctc aggacttatg ttgacccaca tacatatgaa gaccctaccc aagctgttca
     2941 tgagtttgcc aaggaattgg atgccaccaa catatccatt gataaagttg ttggagcagg
     3001 tgaatttgga gaggtgtgca gtggtcgctt aaaacttcct tcaaaaaaag agatttcagt
     3061 ggccattaag accctgaaag ttggctacac agaaaagcag aggagagact tcctgggaga
     3121 agcaagcatt atgggacagt ttgaccaccc caatatcatt cgactggaag gagttgttac
     3181 caaaagtaag ccagttatga ttgtcacaga atacatggag aatggttcct tggatagttt
     3241 cctacgtaaa cacgatgccc agtttactgt cattcagcta gtggggatgc ttcgagggat
     3301 agcatctggc atgaagtacc tgtcagacat gggctatgtt caccgagacc tcgctgctcg
     3361 gaacatcttg atcaacagta acttggtgtg taaggtttct gatttcggac tttcgcgtgt
     3421 cctggaggat gacccagaag ctgcttatac aacaagagga gggaagatcc caatcaggtg
     3481 gacatcacca gaagctatag cctaccgcaa gttcacgtca gccagcgatg tatggagtta
     3541 tgggattgtt ctctgggagg tgatgtctta tggagagaga ccatactggg agatgtccaa
     3601 tcaggatgta attaaagctg tagatgaggg ctatcgactg ccacccccca tggactgccc
     3661 agctgccttg tatcagctga tgctggactg ctggcagaaa gacaggaaca acagacccaa
     3721 gtttgagcag attgttagta ttctggacaa gcttatccgg aatcccggca gcctgaagat
     3781 catcaccagt gcagccgcaa ggccatcaaa ccttcttctg gaccaaagca atgtggatat
     3841 cactaccttc cgcacaacag gtgactggct taatggtgtc tggacagcac actgcaagga
     3901 aatcttcacg ggtgtggagt acagttcttg tgacacaata gccaagattt ccacagatga
     3961 catgaaaaag gttggtgtca ccgtggttgg gccacagaag aagatcatca gtagcattaa
     4021 agctctagag ggcccgcggt tcgaacaaaa actcatctca gaagaggatc tgaatatgca
     4081 taccggtcat catcaccatc accattgagt ttaaacccgc tgatcagcct cgactgtgcc
     4141 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     4201 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     4261 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     4321 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     4381 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     4441 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     4501 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     4561 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     4621 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4681 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     4741 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     4801 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     4861 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4921 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4981 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     5041 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     5101 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     5161 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     5221 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     5281 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     5341 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     5401 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     5461 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     5521 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     5581 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     5641 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     5701 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     5761 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     5821 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     5881 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     5941 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     6001 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat
     6061 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     6121 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     6181 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     6241 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     6301 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     6361 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     6421 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     6481 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     6541 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     6601 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     6661 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     6721 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     6781 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     6841 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     6901 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     6961 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     7021 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     7081 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     7141 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     7201 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     7261 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     7321 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     7381 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     7441 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     7501 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     7561 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     7621 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     7681 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     7741 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     7801 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     7861 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     7921 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     7981 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     8041 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     8101 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     8161 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     8221 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     8281 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     8341 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     8401 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     8461 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     8521 aaaagtgcca cctgacgtc
//
