LOCUS       sequence-198411-        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: D806N EPHA3 pcDNA3.1
FEATURES             Location/Qualifiers
     source          1..8539
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     73..94
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(213..233)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        218..547
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      398..533
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     460..479
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             614..1408
                     /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(668..687)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1278..1297
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1584..1705
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1621..1640)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1675..1694
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1754..1770)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1754..1770)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1767..1789)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1778..1794
                     /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(1802..1808,1809..1826,1827..1832))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1847..1868
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1985..2002)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2156..2744)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2236..2255)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2915..3706,3707..3775))
                     /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     3538..3557
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3776..3880)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3955..3974)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4146..4525
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4526..4729
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4680..4700
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4774..4793
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4774..4792
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             7954..7983
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             7999..8016
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(8039..8056)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8045..8269
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(8315..8539,1..204)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8402..8421)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
       61 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
      121 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
      181 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
      241 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
      301 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
      361 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
      421 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
      481 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
      541 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg
      601 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
      661 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
      721 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
      781 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
      841 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
      901 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
      961 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     1021 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     1081 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     1141 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     1201 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     1261 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     1321 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     1381 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc
     1441 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     1501 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     1561 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     1621 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     1681 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     1741 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     1801 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     1861 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     1921 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     1981 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     2041 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     2101 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     2161 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     2221 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     2281 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     2341 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     2401 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     2461 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     2521 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     2581 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     2641 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     2701 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     2761 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     2821 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     2881 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     2941 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     3001 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     3061 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     3121 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     3181 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     3241 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     3301 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     3361 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     3421 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     3481 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     3541 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     3601 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     3661 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     3721 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     3781 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     3841 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     3901 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat
     3961 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
     4021 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
     4081 gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac
     4141 gcgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
     4201 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     4261 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     4321 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     4381 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     4441 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     4501 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
     4561 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     4621 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     4681 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac
     4741 tagagaaccc actgcttact ggcttatcga aattaatacg actcactata gggagaccca
     4801 agctggctag ttaagcttgg taccgagctc ggatccacta gtccagtgtg gtggaattcg
     4861 tcgagctcta gcgaggtgac agcgtagaac caggagtgtc aaacttgaca tcagcctgcg
     4921 agcggagcat ggtaacttct ccagcaatca gagcgctccc cctcacatca gtggcatgct
     4981 tcatggagat atgctcctct cactgccctc tgcaccagca acatggattg tcagctctcc
     5041 atcctcctcc ttctcagctg ctctgttctc gacagcttcg gggaactgat tccgcagcct
     5101 tccaatgaag tcaatctact ggattcaaaa acaattcaag gggagctggg ctggatctct
     5161 tatccatcac atgggtggga agagatcagt ggtgtggatg aacattacac acccatcagg
     5221 acttaccagg tgtgcaatgt catggaccac agtcaaaaca attggctgag aacaaactgg
     5281 gtccccagga actcagctca gaagatttat gtggagctca agttcactct acgagactgc
     5341 aatagcattc cattggtttt aggaacttgc aaggagacat tcaacctgta ctacatggag
     5401 tctgatgatg atcatggggt gaaatttcga gagcatcagt ttacaaagat tgacaccatt
     5461 gcagctgatg aaagtttcac tcaaatggat cttggggacc gtattctgaa gctcaacact
     5521 gagattagag aagtaggtcc tgtcaacaag aagggatttt atttggcatt tcaagatgtt
     5581 ggtgcttgtg ttgccttggt gtctgtgaga gtatacttca aaaagtgccc atttacagtg
     5641 aagaatctgg ctatgtttcc agacacggta cccatggact cccagtccct ggtggaggtt
     5701 agagggtctt gtgtcaacaa ttctaaggag gaagatcctc caaggatgta ctgcagtaca
     5761 gaaggcgaat ggcttgtacc cattggcaag tgttcctgca atgctggcta tgaagaaaga
     5821 ggttttatgt gccaagcttg tcgaccaggt ttctacaagg cattggatgg taatatgaag
     5881 tgtgctaagt gcccgcctca cagttctact caggaagatg gttcaatgaa ctgcaggtgt
     5941 gagaataatt acttccgggc agacaaagac cctccatcca tggcttgtac ccgacctcca
     6001 tcttcaccaa gaaatgttat ctctaatata aacgagacct cagttatcct ggactggagt
     6061 tggcccctgg acacaggagg ccggaaagat gttaccttca acatcatatg taaaaaatgt
     6121 gggtggaata taaaacagtg tgagccatgc agcccaaatg tccgcttcct ccctcgacag
     6181 tttggactca ccaacaccac ggtgacagtg acagaccttc tggcacatac taactacacc
     6241 tttgagattg atgccgttaa tggggtgtca gagctgagct ccccaccaag acagtttgct
     6301 gcggtcagca tcacaactaa tcaggctgct ccatcacctg tcctgacgat taagaaagat
     6361 cggacctcca gaaatagcat ctctttgtcc tggcaagaac ctgaacatcc taatgggatc
     6421 atattggact acgaggtcaa atactatgaa aagcaggaac aagaaacaag ttataccatt
     6481 ctgagggcaa gaggcacaaa tgttaccatc agtagcctca agcctgacac tatatacgta
     6541 ttccaaatcc gagcccgaac agccgctgga tatgggacga acagccgcaa gtttgagttt
     6601 gaaactagtc cagactcttt ctccatctct ggtgaaagta gccaagtggt catgatcgcc
     6661 atttcagcgg cagtagcaat tattctcctc actgttgtca tctatgtttt gattgggagg
     6721 ttctgtggct ataagtcaaa acatggggca gatgaaaaaa gacttcattt tggcaatggg
     6781 catttaaaac ttccaggtct caggacttat gttgacccac atacatatga agaccctacc
     6841 caagctgttc atgagtttgc caaggaattg gatgccacca acatatccat tgataaagtt
     6901 gttggagcag gtgaatttgg agaggtgtgc agtggtcgct taaaacttcc ttcaaaaaaa
     6961 gagatttcag tggccattaa gaccctgaaa gttggctaca cagaaaagca gaggagagac
     7021 ttcctgggag aagcaagcat tatgggacag tttgaccacc ccaatatcat tcgactggaa
     7081 ggagttgtta ccaaaagtaa gccagttatg attgtcacag aatacatgga gaatggttcc
     7141 ttggatagtt tcctacgtaa acacgatgcc cagtttactg tcattcagct agtggggatg
     7201 cttcgaggga tagcatctgg catgaagtac ctgtcagaca tgggctatgt tcaccgagac
     7261 ctcgctgctc ggaacatctt gatcaacagt aacttggtgt gtaaggtttc tgatttcgga
     7321 ctttcgcgtg tcctggagga tgacccagaa gctgcttata caacaagagg agggaagatc
     7381 ccaatcaggt ggacatcacc agaagctata gcctaccgca agttcacgtc agccagcaat
     7441 gtatggagtt atgggattgt tctctgggag gtgatgtctt atggagagag accatactgg
     7501 gagatgtcca atcaggatgt aattaaagct gtagatgagg gctatcgact gccacccccc
     7561 atggactgcc cagctgcctt gtatcagctg atgctggact gctggcagaa agacaggaac
     7621 aacagaccca agtttgagca gattgttagt attctggaca agcttatccg gaatcccggc
     7681 agcctgaaga tcatcaccag tgcagccgca aggccatcaa accttcttct ggaccaaagc
     7741 aatgtggata tcactacctt ccgcacaaca ggtgactggc ttaatggtgt ctggacagca
     7801 cactgcaagg aaatcttcac gggtgtggag tacagttctt gtgacacaat agccaagatt
     7861 tccacagatg acatgaaaaa ggttggtgtc accgtggttg ggccacagaa gaagatcatc
     7921 agtagcatta aagctctaga gggcccgcgg ttcgaacaaa aactcatctc agaagaggat
     7981 ctgaatatgc ataccggtca tcatcaccat caccattgag tttaaacccg ctgatcagcc
     8041 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     8101 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     8161 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     8221 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     8281 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     8341 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     8401 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     8461 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     8521 aaacttgatt agggtgatg
//
