LOCUS       sequence-198418-        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: T660K EPHA3 pcDNA3.1
FEATURES             Location/Qualifiers
     source          1..8539
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      96..231
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     158..177
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             312..1106
                     /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(366..385)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     976..995
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1282..1403
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1319..1338)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1373..1392
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1452..1468)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1452..1468)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1465..1487)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1476..1492
                     /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(1500..1506,1507..1524,1525..1530))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1545..1566
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1683..1700)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1854..2442)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1934..1953)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2613..3404,3405..3473))
                     /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     3236..3255
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3474..3578)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3653..3672)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3844..4223
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4224..4427
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4378..4398
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4472..4491
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4472..4490
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             7652..7681
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             7697..7714
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7737..7754)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7743..7967
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8013..8441
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8100..8119)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8310..8331
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8450..8470)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(8455..8539,1..245)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
ORIGIN
        1 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
       61 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
      121 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
      181 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
      241 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
      301 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
      361 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
      421 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
      481 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
      541 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
      601 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
      661 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
      721 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
      781 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
      841 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
      901 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
      961 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     1021 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     1081 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa
     1141 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     1201 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     1261 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     1321 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     1381 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     1441 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     1501 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     1561 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     1621 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     1681 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     1741 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     1801 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     1861 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     1921 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     1981 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2041 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2101 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2161 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     2221 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     2281 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     2341 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     2401 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     2461 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     2521 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     2581 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     2641 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     2701 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     2761 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     2821 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     2881 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     2941 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     3001 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     3061 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     3121 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     3181 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     3241 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     3301 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     3361 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     3421 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     3481 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     3541 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     3601 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
     3661 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     3721 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
     3781 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc
     3841 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
     3901 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     3961 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     4021 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     4081 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     4141 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     4201 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
     4261 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
     4321 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
     4381 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta
     4441 gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagacccaag
     4501 ctggctagtt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgtc
     4561 gagctctagc gaggtgacag cgtagaacca ggagtgtcaa acttgacatc agcctgcgag
     4621 cggagcatgg taacttctcc agcaatcaga gcgctccccc tcacatcagt ggcatgcttc
     4681 atggagatat gctcctctca ctgccctctg caccagcaac atggattgtc agctctccat
     4741 cctcctcctt ctcagctgct ctgttctcga cagcttcggg gaactgattc cgcagccttc
     4801 caatgaagtc aatctactgg attcaaaaac aattcaaggg gagctgggct ggatctctta
     4861 tccatcacat gggtgggaag agatcagtgg tgtggatgaa cattacacac ccatcaggac
     4921 ttaccaggtg tgcaatgtca tggaccacag tcaaaacaat tggctgagaa caaactgggt
     4981 ccccaggaac tcagctcaga agatttatgt ggagctcaag ttcactctac gagactgcaa
     5041 tagcattcca ttggttttag gaacttgcaa ggagacattc aacctgtact acatggagtc
     5101 tgatgatgat catggggtga aatttcgaga gcatcagttt acaaagattg acaccattgc
     5161 agctgatgaa agtttcactc aaatggatct tggggaccgt attctgaagc tcaacactga
     5221 gattagagaa gtaggtcctg tcaacaagaa gggattttat ttggcatttc aagatgttgg
     5281 tgcttgtgtt gccttggtgt ctgtgagagt atacttcaaa aagtgcccat ttacagtgaa
     5341 gaatctggct atgtttccag acacggtacc catggactcc cagtccctgg tggaggttag
     5401 agggtcttgt gtcaacaatt ctaaggagga agatcctcca aggatgtact gcagtacaga
     5461 aggcgaatgg cttgtaccca ttggcaagtg ttcctgcaat gctggctatg aagaaagagg
     5521 ttttatgtgc caagcttgtc gaccaggttt ctacaaggca ttggatggta atatgaagtg
     5581 tgctaagtgc ccgcctcaca gttctactca ggaagatggt tcaatgaact gcaggtgtga
     5641 gaataattac ttccgggcag acaaagaccc tccatccatg gcttgtaccc gacctccatc
     5701 ttcaccaaga aatgttatct ctaatataaa cgagacctca gttatcctgg actggagttg
     5761 gcccctggac acaggaggcc ggaaagatgt taccttcaac atcatatgta aaaaatgtgg
     5821 gtggaatata aaacagtgtg agccatgcag cccaaatgtc cgcttcctcc ctcgacagtt
     5881 tggactcacc aacaccacgg tgacagtgac agaccttctg gcacatacta actacacctt
     5941 tgagattgat gccgttaatg gggtgtcaga gctgagctcc ccaccaagac agtttgctgc
     6001 ggtcagcatc acaactaatc aggctgctcc atcacctgtc ctgacgatta agaaagatcg
     6061 gacctccaga aatagcatct ctttgtcctg gcaagaacct gaacatccta atgggatcat
     6121 attggactac gaggtcaaat actatgaaaa gcaggaacaa gaaacaagtt ataccattct
     6181 gagggcaaga ggcacaaatg ttaccatcag tagcctcaag cctgacacta tatacgtatt
     6241 ccaaatccga gcccgaacag ccgctggata tgggacgaac agccgcaagt ttgagtttga
     6301 aactagtcca gactctttct ccatctctgg tgaaagtagc caagtggtca tgatcgccat
     6361 ttcagcggca gtagcaatta ttctcctcac tgttgtcatc tatgttttga ttgggaggtt
     6421 ctgtggctat aagtcaaaac atggggcaga tgaaaaaaga cttcattttg gcaatgggca
     6481 tttaaaactt ccaggtctca ggacttatgt tgacccacat acatatgaag accctaccca
     6541 agctgttcat gagtttgcca aggaattgga tgccaccaac atatccattg ataaagttgt
     6601 tggagcaggt gaatttggag aggtgtgcag tggtcgctta aaacttcctt caaaaaaaga
     6661 gatttcagtg gccattaaga ccctgaaagt tggctacaaa gaaaagcaga ggagagactt
     6721 cctgggagaa gcaagcatta tgggacagtt tgaccacccc aatatcattc gactggaagg
     6781 agttgttacc aaaagtaagc cagttatgat tgtcacagaa tacatggaga atggttcctt
     6841 ggatagtttc ctacgtaaac acgatgccca gtttactgtc attcagctag tggggatgct
     6901 tcgagggata gcatctggca tgaagtacct gtcagacatg ggctatgttc accgagacct
     6961 cgctgctcgg aacatcttga tcaacagtaa cttggtgtgt aaggtttctg atttcggact
     7021 ttcgcgtgtc ctggaggatg acccagaagc tgcttataca acaagaggag ggaagatccc
     7081 aatcaggtgg acatcaccag aagctatagc ctaccgcaag ttcacgtcag ccagcgatgt
     7141 atggagttat gggattgttc tctgggaggt gatgtcttat ggagagagac catactggga
     7201 gatgtccaat caggatgtaa ttaaagctgt agatgagggc tatcgactgc caccccccat
     7261 ggactgccca gctgccttgt atcagctgat gctggactgc tggcagaaag acaggaacaa
     7321 cagacccaag tttgagcaga ttgttagtat tctggacaag cttatccgga atcccggcag
     7381 cctgaagatc atcaccagtg cagccgcaag gccatcaaac cttcttctgg accaaagcaa
     7441 tgtggatatc actaccttcc gcacaacagg tgactggctt aatggtgtct ggacagcaca
     7501 ctgcaaggaa atcttcacgg gtgtggagta cagttcttgt gacacaatag ccaagatttc
     7561 cacagatgac atgaaaaagg ttggtgtcac cgtggttggg ccacagaaga agatcatcag
     7621 tagcattaaa gctctagagg gcccgcggtt cgaacaaaaa ctcatctcag aagaggatct
     7681 gaatatgcat accggtcatc atcaccatca ccattgagtt taaacccgct gatcagcctc
     7741 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     7801 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     7861 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     7921 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     7981 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     8041 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     8101 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     8161 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     8221 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     8281 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     8341 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     8401 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     8461 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     8521 ctcaattagt cagcaacca
//
