LOCUS       sequence-199335-        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: D678E EPHA3 pcDNA3.1
FEATURES             Location/Qualifiers
     source          1..8539
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      237..665
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(324..343)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     534..555
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(674..694)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        679..1008
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      859..994
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     921..940
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1075..1869
                     /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(1129..1148)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1739..1758
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2045..2166
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2082..2101)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2136..2155
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2215..2231)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2215..2231)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2228..2250)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2239..2255
                     /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(2263..2269,2270..2287,2288..2293))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2308..2329
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2446..2463)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2617..3205)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2697..2716)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3376..4167,4168..4236))
                     /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     3999..4018
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4237..4341)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4416..4435)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4607..4986
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4987..5190
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5141..5161
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5235..5254
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5235..5253
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             8415..8444
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             8460..8477
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(8500..8517)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    join(8506..8539,1..191)
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
       61 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
      121 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
      181 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
      241 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
      301 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
      361 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
      421 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
      481 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
      541 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
      601 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
      661 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
      721 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
      781 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
      841 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
      901 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
      961 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     1021 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     1081 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     1141 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     1201 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     1261 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     1321 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     1381 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     1441 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     1501 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     1561 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     1621 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     1681 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     1741 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     1801 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     1861 ttcttctgag cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc
     1921 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     1981 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     2041 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     2101 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     2161 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     2221 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     2281 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     2341 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     2401 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     2461 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     2521 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     2581 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2641 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2701 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2761 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2821 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2881 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2941 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3001 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     3061 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3121 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3181 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3241 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3301 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3361 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     3421 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     3481 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     3541 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     3601 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     3661 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     3721 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     3781 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     3841 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     3901 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     3961 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     4021 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     4081 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     4141 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     4201 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     4261 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     4321 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     4381 gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag
     4441 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
     4501 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt
     4561 aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca ttgattattg
     4621 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
     4681 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
     4741 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
     4801 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
     4861 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
     4921 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
     4981 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
     5041 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
     5101 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
     5161 gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc cactgcttac
     5221 tggcttatcg aaattaatac gactcactat agggagaccc aagctggcta gttaagcttg
     5281 gtaccgagct cggatccact agtccagtgt ggtggaattc gtcgagctct agcgaggtga
     5341 cagcgtagaa ccaggagtgt caaacttgac atcagcctgc gagcggagca tggtaacttc
     5401 tccagcaatc agagcgctcc ccctcacatc agtggcatgc ttcatggaga tatgctcctc
     5461 tcactgccct ctgcaccagc aacatggatt gtcagctctc catcctcctc cttctcagct
     5521 gctctgttct cgacagcttc ggggaactga ttccgcagcc ttccaatgaa gtcaatctac
     5581 tggattcaaa aacaattcaa ggggagctgg gctggatctc ttatccatca catgggtggg
     5641 aagagatcag tggtgtggat gaacattaca cacccatcag gacttaccag gtgtgcaatg
     5701 tcatggacca cagtcaaaac aattggctga gaacaaactg ggtccccagg aactcagctc
     5761 agaagattta tgtggagctc aagttcactc tacgagactg caatagcatt ccattggttt
     5821 taggaacttg caaggagaca ttcaacctgt actacatgga gtctgatgat gatcatgggg
     5881 tgaaatttcg agagcatcag tttacaaaga ttgacaccat tgcagctgat gaaagtttca
     5941 ctcaaatgga tcttggggac cgtattctga agctcaacac tgagattaga gaagtaggtc
     6001 ctgtcaacaa gaagggattt tatttggcat ttcaagatgt tggtgcttgt gttgccttgg
     6061 tgtctgtgag agtatacttc aaaaagtgcc catttacagt gaagaatctg gctatgtttc
     6121 cagacacggt acccatggac tcccagtccc tggtggaggt tagagggtct tgtgtcaaca
     6181 attctaagga ggaagatcct ccaaggatgt actgcagtac agaaggcgaa tggcttgtac
     6241 ccattggcaa gtgttcctgc aatgctggct atgaagaaag aggttttatg tgccaagctt
     6301 gtcgaccagg tttctacaag gcattggatg gtaatatgaa gtgtgctaag tgcccgcctc
     6361 acagttctac tcaggaagat ggttcaatga actgcaggtg tgagaataat tacttccggg
     6421 cagacaaaga ccctccatcc atggcttgta cccgacctcc atcttcacca agaaatgtta
     6481 tctctaatat aaacgagacc tcagttatcc tggactggag ttggcccctg gacacaggag
     6541 gccggaaaga tgttaccttc aacatcatat gtaaaaaatg tgggtggaat ataaaacagt
     6601 gtgagccatg cagcccaaat gtccgcttcc tccctcgaca gtttggactc accaacacca
     6661 cggtgacagt gacagacctt ctggcacata ctaactacac ctttgagatt gatgccgtta
     6721 atggggtgtc agagctgagc tccccaccaa gacagtttgc tgcggtcagc atcacaacta
     6781 atcaggctgc tccatcacct gtcctgacga ttaagaaaga tcggacctcc agaaatagca
     6841 tctctttgtc ctggcaagaa cctgaacatc ctaatgggat catattggac tacgaggtca
     6901 aatactatga aaagcaggaa caagaaacaa gttataccat tctgagggca agaggcacaa
     6961 atgttaccat cagtagcctc aagcctgaca ctatatacgt attccaaatc cgagcccgaa
     7021 cagccgctgg atatgggacg aacagccgca agtttgagtt tgaaactagt ccagactctt
     7081 tctccatctc tggtgaaagt agccaagtgg tcatgatcgc catttcagcg gcagtagcaa
     7141 ttattctcct cactgttgtc atctatgttt tgattgggag gttctgtggc tataagtcaa
     7201 aacatggggc agatgaaaaa agacttcatt ttggcaatgg gcatttaaaa cttccaggtc
     7261 tcaggactta tgttgaccca catacatatg aagaccctac ccaagctgtt catgagtttg
     7321 ccaaggaatt ggatgccacc aacatatcca ttgataaagt tgttggagca ggtgaatttg
     7381 gagaggtgtg cagtggtcgc ttaaaacttc cttcaaaaaa agagatttca gtggccatta
     7441 agaccctgaa agttggctac acagaaaagc agaggagaga cttcctggga gaagcaagca
     7501 ttatgggaca gtttgaacac cccaatatca ttcgactgga aggagttgtt accaaaagta
     7561 agccagttat gattgtcaca gaatacatgg agaatggttc cttggatagt ttcctacgta
     7621 aacacgatgc ccagtttact gtcattcagc tagtggggat gcttcgaggg atagcatctg
     7681 gcatgaagta cctgtcagac atgggctatg ttcaccgaga cctcgctgct cggaacatct
     7741 tgatcaacag taacttggtg tgtaaggttt ctgatttcgg actttcgcgt gtcctggagg
     7801 atgacccaga agctgcttat acaacaagag gagggaagat cccaatcagg tggacatcac
     7861 cagaagctat agcctaccgc aagttcacgt cagccagcga tgtatggagt tatgggattg
     7921 ttctctggga ggtgatgtct tatggagaga gaccatactg ggagatgtcc aatcaggatg
     7981 taattaaagc tgtagatgag ggctatcgac tgccaccccc catggactgc ccagctgcct
     8041 tgtatcagct gatgctggac tgctggcaga aagacaggaa caacagaccc aagtttgagc
     8101 agattgttag tattctggac aagcttatcc ggaatcccgg cagcctgaag atcatcacca
     8161 gtgcagccgc aaggccatca aaccttcttc tggaccaaag caatgtggat atcactacct
     8221 tccgcacaac aggtgactgg cttaatggtg tctggacagc acactgcaag gaaatcttca
     8281 cgggtgtgga gtacagttct tgtgacacaa tagccaagat ttccacagat gacatgaaaa
     8341 aggttggtgt caccgtggtt gggccacaga agaagatcat cagtagcatt aaagctctag
     8401 agggcccgcg gttcgaacaa aaactcatct cagaagagga tctgaatatg cataccggtc
     8461 atcatcacca tcaccattga gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
     8521 gccagccatc tgttgtttg
//
