LOCUS       sequence-198417-        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: S449F EPHA3 pcDNA3.1
FEATURES             Location/Qualifiers
     source          1..8539
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     148..167
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    454..575
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(491..510)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     545..564
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(624..640)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(624..640)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(637..659)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    648..664
                     /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(672..678,679..696,697..702))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    717..738
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(855..872)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1026..1614)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1106..1125)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1785..2576,2577..2645))
                     /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     2408..2427
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2646..2750)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2825..2844)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3016..3395
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3396..3599
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3550..3570
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3644..3663
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3644..3662
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6824..6853
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             6869..6886
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6909..6926)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6915..7139
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7185..7613
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7272..7291)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7482..7503
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7622..7642)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7627..7956
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7807..7942
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7869..7888
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             join(8023..8539,1..278)
                     /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(8077..8096)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
       61 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
      121 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
      181 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
      241 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgcgaa
      301 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
      361 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
      421 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
      481 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
      541 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
      601 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
      661 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
      721 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
      781 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
      841 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
      901 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
      961 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     1021 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     1081 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     1141 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     1201 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     1261 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     1321 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     1381 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     1441 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     1501 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     1561 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     1621 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     1681 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     1741 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     1801 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     1861 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     1921 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     1981 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     2041 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     2101 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     2161 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     2221 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     2281 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     2341 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     2401 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     2461 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     2521 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     2581 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     2641 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     2701 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     2761 cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc
     2821 tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt
     2881 tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg
     2941 acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg
     3001 ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca attacggggt
     3061 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     3121 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     3181 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     3241 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     3301 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     3361 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
     3421 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
     3481 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
     3541 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc
     3601 tctggctaac tagagaaccc actgcttact ggcttatcga aattaatacg actcactata
     3661 gggagaccca agctggctag ttaagcttgg taccgagctc ggatccacta gtccagtgtg
     3721 gtggaattcg tcgagctcta gcgaggtgac agcgtagaac caggagtgtc aaacttgaca
     3781 tcagcctgcg agcggagcat ggtaacttct ccagcaatca gagcgctccc cctcacatca
     3841 gtggcatgct tcatggagat atgctcctct cactgccctc tgcaccagca acatggattg
     3901 tcagctctcc atcctcctcc ttctcagctg ctctgttctc gacagcttcg gggaactgat
     3961 tccgcagcct tccaatgaag tcaatctact ggattcaaaa acaattcaag gggagctggg
     4021 ctggatctct tatccatcac atgggtggga agagatcagt ggtgtggatg aacattacac
     4081 acccatcagg acttaccagg tgtgcaatgt catggaccac agtcaaaaca attggctgag
     4141 aacaaactgg gtccccagga actcagctca gaagatttat gtggagctca agttcactct
     4201 acgagactgc aatagcattc cattggtttt aggaacttgc aaggagacat tcaacctgta
     4261 ctacatggag tctgatgatg atcatggggt gaaatttcga gagcatcagt ttacaaagat
     4321 tgacaccatt gcagctgatg aaagtttcac tcaaatggat cttggggacc gtattctgaa
     4381 gctcaacact gagattagag aagtaggtcc tgtcaacaag aagggatttt atttggcatt
     4441 tcaagatgtt ggtgcttgtg ttgccttggt gtctgtgaga gtatacttca aaaagtgccc
     4501 atttacagtg aagaatctgg ctatgtttcc agacacggta cccatggact cccagtccct
     4561 ggtggaggtt agagggtctt gtgtcaacaa ttctaaggag gaagatcctc caaggatgta
     4621 ctgcagtaca gaaggcgaat ggcttgtacc cattggcaag tgttcctgca atgctggcta
     4681 tgaagaaaga ggttttatgt gccaagcttg tcgaccaggt ttctacaagg cattggatgg
     4741 taatatgaag tgtgctaagt gcccgcctca cagttctact caggaagatg gttcaatgaa
     4801 ctgcaggtgt gagaataatt acttccgggc agacaaagac cctccatcca tggcttgtac
     4861 ccgacctcca tcttcaccaa gaaatgttat ctctaatata aacgagacct cagttatcct
     4921 ggactggagt tggcccctgg acacaggagg ccggaaagat gttaccttca acatcatatg
     4981 taaaaaatgt gggtggaata taaaacagtg tgagccatgc agcccaaatg tccgcttcct
     5041 ccctcgacag tttggactca ccaacaccac ggtgacagtg acagaccttc tggcacatac
     5101 taactacacc tttgagattg atgccgttaa tggggtgtca gagctgagct ccccaccaag
     5161 acagtttgct gcggtcagca tcacaactaa tcaggctgct ccatcacctg tcctgacgat
     5221 taagaaagat cggaccttca gaaatagcat ctctttgtcc tggcaagaac ctgaacatcc
     5281 taatgggatc atattggact acgaggtcaa atactatgaa aagcaggaac aagaaacaag
     5341 ttataccatt ctgagggcaa gaggcacaaa tgttaccatc agtagcctca agcctgacac
     5401 tatatacgta ttccaaatcc gagcccgaac agccgctgga tatgggacga acagccgcaa
     5461 gtttgagttt gaaactagtc cagactcttt ctccatctct ggtgaaagta gccaagtggt
     5521 catgatcgcc atttcagcgg cagtagcaat tattctcctc actgttgtca tctatgtttt
     5581 gattgggagg ttctgtggct ataagtcaaa acatggggca gatgaaaaaa gacttcattt
     5641 tggcaatggg catttaaaac ttccaggtct caggacttat gttgacccac atacatatga
     5701 agaccctacc caagctgttc atgagtttgc caaggaattg gatgccacca acatatccat
     5761 tgataaagtt gttggagcag gtgaatttgg agaggtgtgc agtggtcgct taaaacttcc
     5821 ttcaaaaaaa gagatttcag tggccattaa gaccctgaaa gttggctaca cagaaaagca
     5881 gaggagagac ttcctgggag aagcaagcat tatgggacag tttgaccacc ccaatatcat
     5941 tcgactggaa ggagttgtta ccaaaagtaa gccagttatg attgtcacag aatacatgga
     6001 gaatggttcc ttggatagtt tcctacgtaa acacgatgcc cagtttactg tcattcagct
     6061 agtggggatg cttcgaggga tagcatctgg catgaagtac ctgtcagaca tgggctatgt
     6121 tcaccgagac ctcgctgctc ggaacatctt gatcaacagt aacttggtgt gtaaggtttc
     6181 tgatttcgga ctttcgcgtg tcctggagga tgacccagaa gctgcttata caacaagagg
     6241 agggaagatc ccaatcaggt ggacatcacc agaagctata gcctaccgca agttcacgtc
     6301 agccagcgat gtatggagtt atgggattgt tctctgggag gtgatgtctt atggagagag
     6361 accatactgg gagatgtcca atcaggatgt aattaaagct gtagatgagg gctatcgact
     6421 gccacccccc atggactgcc cagctgcctt gtatcagctg atgctggact gctggcagaa
     6481 agacaggaac aacagaccca agtttgagca gattgttagt attctggaca agcttatccg
     6541 gaatcccggc agcctgaaga tcatcaccag tgcagccgca aggccatcaa accttcttct
     6601 ggaccaaagc aatgtggata tcactacctt ccgcacaaca ggtgactggc ttaatggtgt
     6661 ctggacagca cactgcaagg aaatcttcac gggtgtggag tacagttctt gtgacacaat
     6721 agccaagatt tccacagatg acatgaaaaa ggttggtgtc accgtggttg ggccacagaa
     6781 gaagatcatc agtagcatta aagctctaga gggcccgcgg ttcgaacaaa aactcatctc
     6841 agaagaggat ctgaatatgc ataccggtca tcatcaccat caccattgag tttaaacccg
     6901 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     6961 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     7021 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     7081 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     7141 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     7201 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     7261 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     7321 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     7381 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     7441 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     7501 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     7561 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     7621 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     7681 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     7741 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     7801 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     7861 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     7921 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     7981 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     8041 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     8101 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     8161 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     8221 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     8281 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     8341 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     8401 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     8461 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     8521 ccagccgaac tgttcgcca
//
