LOCUS       sequence-198406-        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: N85S EPHA3 pcDNA3.1
FEATURES             Location/Qualifiers
     source          1..8539
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      113..248
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     175..194
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             329..1123
                     /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(383..402)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     993..1012
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1299..1420
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1336..1355)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1390..1409
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1469..1485)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1469..1485)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1482..1504)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1493..1509
                     /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(1517..1523,1524..1541,1542..1547))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1562..1583
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1700..1717)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1871..2459)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1951..1970)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2630..3421,3422..3490))
                     /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     3253..3272
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3491..3595)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3670..3689)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3861..4240
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4241..4444
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4395..4415
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4489..4508
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4489..4507
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             7669..7698
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             7714..7731
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7754..7771)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7760..7984
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8030..8458
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8117..8136)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8327..8348
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8467..8487)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(8472..8539,1..262)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
ORIGIN
        1 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
       61 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
      121 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
      181 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
      241 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
      301 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
      361 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
      421 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
      481 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
      541 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
      601 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
      661 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
      721 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
      781 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
      841 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
      901 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
      961 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     1021 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     1081 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     1141 gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     1201 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     1261 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     1321 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     1381 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     1441 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     1501 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     1561 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     1621 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     1681 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     1741 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     1801 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     1861 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     1921 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     1981 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     2041 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     2101 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     2161 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     2221 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     2281 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     2341 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     2401 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     2461 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     2521 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     2581 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     2641 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     2701 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     2761 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     2821 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     2881 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     2941 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     3001 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     3061 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     3121 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     3181 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     3241 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     3301 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     3361 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     3421 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     3481 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     3541 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     3601 ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg
     3661 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg
     3721 tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt
     3781 gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt
     3841 acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac
     3901 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     3961 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     4021 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     4081 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     4141 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     4201 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
     4261 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     4321 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
     4381 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
     4441 agctctctgg ctaactagag aacccactgc ttactggctt atcgaaatta atacgactca
     4501 ctatagggag acccaagctg gctagttaag cttggtaccg agctcggatc cactagtcca
     4561 gtgtggtgga attcgtcgag ctctagcgag gtgacagcgt agaaccagga gtgtcaaact
     4621 tgacatcagc ctgcgagcgg agcatggtaa cttctccagc aatcagagcg ctccccctca
     4681 catcagtggc atgcttcatg gagatatgct cctctcactg ccctctgcac cagcaacatg
     4741 gattgtcagc tctccatcct cctccttctc agctgctctg ttctcgacag cttcggggaa
     4801 ctgattccgc agccttccaa tgaagtcaat ctactggatt caaaaacaat tcaaggggag
     4861 ctgggctgga tctcttatcc atcacatggg tgggaagaga tcagtggtgt ggatgaacat
     4921 tacacaccca tcaggactta ccaggtgtgc aatgtcatgg accacagtca aaacaattgg
     4981 ctgagaacaa gctgggtccc caggaactca gctcagaaga tttatgtgga gctcaagttc
     5041 actctacgag actgcaatag cattccattg gttttaggaa cttgcaagga gacattcaac
     5101 ctgtactaca tggagtctga tgatgatcat ggggtgaaat ttcgagagca tcagtttaca
     5161 aagattgaca ccattgcagc tgatgaaagt ttcactcaaa tggatcttgg ggaccgtatt
     5221 ctgaagctca acactgagat tagagaagta ggtcctgtca acaagaaggg attttatttg
     5281 gcatttcaag atgttggtgc ttgtgttgcc ttggtgtctg tgagagtata cttcaaaaag
     5341 tgcccattta cagtgaagaa tctggctatg tttccagaca cggtacccat ggactcccag
     5401 tccctggtgg aggttagagg gtcttgtgtc aacaattcta aggaggaaga tcctccaagg
     5461 atgtactgca gtacagaagg cgaatggctt gtacccattg gcaagtgttc ctgcaatgct
     5521 ggctatgaag aaagaggttt tatgtgccaa gcttgtcgac caggtttcta caaggcattg
     5581 gatggtaata tgaagtgtgc taagtgcccg cctcacagtt ctactcagga agatggttca
     5641 atgaactgca ggtgtgagaa taattacttc cgggcagaca aagaccctcc atccatggct
     5701 tgtacccgac ctccatcttc accaagaaat gttatctcta atataaacga gacctcagtt
     5761 atcctggact ggagttggcc cctggacaca ggaggccgga aagatgttac cttcaacatc
     5821 atatgtaaaa aatgtgggtg gaatataaaa cagtgtgagc catgcagccc aaatgtccgc
     5881 ttcctccctc gacagtttgg actcaccaac accacggtga cagtgacaga ccttctggca
     5941 catactaact acacctttga gattgatgcc gttaatgggg tgtcagagct gagctcccca
     6001 ccaagacagt ttgctgcggt cagcatcaca actaatcagg ctgctccatc acctgtcctg
     6061 acgattaaga aagatcggac ctccagaaat agcatctctt tgtcctggca agaacctgaa
     6121 catcctaatg ggatcatatt ggactacgag gtcaaatact atgaaaagca ggaacaagaa
     6181 acaagttata ccattctgag ggcaagaggc acaaatgtta ccatcagtag cctcaagcct
     6241 gacactatat acgtattcca aatccgagcc cgaacagccg ctggatatgg gacgaacagc
     6301 cgcaagtttg agtttgaaac tagtccagac tctttctcca tctctggtga aagtagccaa
     6361 gtggtcatga tcgccatttc agcggcagta gcaattattc tcctcactgt tgtcatctat
     6421 gttttgattg ggaggttctg tggctataag tcaaaacatg gggcagatga aaaaagactt
     6481 cattttggca atgggcattt aaaacttcca ggtctcagga cttatgttga cccacataca
     6541 tatgaagacc ctacccaagc tgttcatgag tttgccaagg aattggatgc caccaacata
     6601 tccattgata aagttgttgg agcaggtgaa tttggagagg tgtgcagtgg tcgcttaaaa
     6661 cttccttcaa aaaaagagat ttcagtggcc attaagaccc tgaaagttgg ctacacagaa
     6721 aagcagagga gagacttcct gggagaagca agcattatgg gacagtttga ccaccccaat
     6781 atcattcgac tggaaggagt tgttaccaaa agtaagccag ttatgattgt cacagaatac
     6841 atggagaatg gttccttgga tagtttccta cgtaaacacg atgcccagtt tactgtcatt
     6901 cagctagtgg ggatgcttcg agggatagca tctggcatga agtacctgtc agacatgggc
     6961 tatgttcacc gagacctcgc tgctcggaac atcttgatca acagtaactt ggtgtgtaag
     7021 gtttctgatt tcggactttc gcgtgtcctg gaggatgacc cagaagctgc ttatacaaca
     7081 agaggaggga agatcccaat caggtggaca tcaccagaag ctatagccta ccgcaagttc
     7141 acgtcagcca gcgatgtatg gagttatggg attgttctct gggaggtgat gtcttatgga
     7201 gagagaccat actgggagat gtccaatcag gatgtaatta aagctgtaga tgagggctat
     7261 cgactgccac cccccatgga ctgcccagct gccttgtatc agctgatgct ggactgctgg
     7321 cagaaagaca ggaacaacag acccaagttt gagcagattg ttagtattct ggacaagctt
     7381 atccggaatc ccggcagcct gaagatcatc accagtgcag ccgcaaggcc atcaaacctt
     7441 cttctggacc aaagcaatgt ggatatcact accttccgca caacaggtga ctggcttaat
     7501 ggtgtctgga cagcacactg caaggaaatc ttcacgggtg tggagtacag ttcttgtgac
     7561 acaatagcca agatttccac agatgacatg aaaaaggttg gtgtcaccgt ggttgggcca
     7621 cagaagaaga tcatcagtag cattaaagct ctagagggcc cgcggttcga acaaaaactc
     7681 atctcagaag aggatctgaa tatgcatacc ggtcatcatc accatcacca ttgagtttaa
     7741 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     7801 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     7861 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     7921 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     7981 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
     8041 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     8101 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     8161 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     8221 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     8281 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     8341 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     8401 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     8461 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     8521 gtatgcaaag catgcatct
//
