LOCUS       sequence-198409-        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: K761N EPHA3 pcDNA3.1
FEATURES             Location/Qualifiers
     source          1..8539
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     55..74
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             209..1003
                     /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(263..282)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     873..892
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1179..1300
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1216..1235)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1270..1289
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1349..1365)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1349..1365)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1362..1384)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1373..1389
                     /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(1397..1403,1404..1421,1422..1427))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1442..1463
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1580..1597)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1751..2339)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1831..1850)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2510..3301,3302..3370))
                     /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     3133..3152
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3371..3475)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3550..3569)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3741..4120
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4121..4324
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4275..4295
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4369..4388
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4369..4387
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             7549..7578
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             7594..7611
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7634..7651)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7640..7864
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7910..8338
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7997..8016)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8207..8228
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8347..8367)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(8352..8539,1..142)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(8532..8539,1..128)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
       61 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
      121 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
      181 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
      241 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
      301 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
      361 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
      421 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
      481 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
      541 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
      601 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
      661 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
      721 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
      781 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
      841 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
      901 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
      961 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     1021 gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     1081 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     1141 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     1201 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     1261 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     1321 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     1381 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     1441 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     1501 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     1561 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     1621 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     1681 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     1741 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     1801 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     1861 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     1921 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     1981 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     2041 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     2101 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     2161 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     2221 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     2281 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     2341 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     2401 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     2461 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     2521 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     2581 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     2641 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     2701 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     2761 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     2821 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     2881 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     2941 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     3001 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     3061 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     3121 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     3181 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     3241 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     3301 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     3361 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     3421 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     3481 ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg
     3541 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg
     3601 tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt
     3661 gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt
     3721 acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac
     3781 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     3841 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     3901 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     3961 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     4021 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     4081 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
     4141 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     4201 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
     4261 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
     4321 agctctctgg ctaactagag aacccactgc ttactggctt atcgaaatta atacgactca
     4381 ctatagggag acccaagctg gctagttaag cttggtaccg agctcggatc cactagtcca
     4441 gtgtggtgga attcgtcgag ctctagcgag gtgacagcgt agaaccagga gtgtcaaact
     4501 tgacatcagc ctgcgagcgg agcatggtaa cttctccagc aatcagagcg ctccccctca
     4561 catcagtggc atgcttcatg gagatatgct cctctcactg ccctctgcac cagcaacatg
     4621 gattgtcagc tctccatcct cctccttctc agctgctctg ttctcgacag cttcggggaa
     4681 ctgattccgc agccttccaa tgaagtcaat ctactggatt caaaaacaat tcaaggggag
     4741 ctgggctgga tctcttatcc atcacatggg tgggaagaga tcagtggtgt ggatgaacat
     4801 tacacaccca tcaggactta ccaggtgtgc aatgtcatgg accacagtca aaacaattgg
     4861 ctgagaacaa actgggtccc caggaactca gctcagaaga tttatgtgga gctcaagttc
     4921 actctacgag actgcaatag cattccattg gttttaggaa cttgcaagga gacattcaac
     4981 ctgtactaca tggagtctga tgatgatcat ggggtgaaat ttcgagagca tcagtttaca
     5041 aagattgaca ccattgcagc tgatgaaagt ttcactcaaa tggatcttgg ggaccgtatt
     5101 ctgaagctca acactgagat tagagaagta ggtcctgtca acaagaaggg attttatttg
     5161 gcatttcaag atgttggtgc ttgtgttgcc ttggtgtctg tgagagtata cttcaaaaag
     5221 tgcccattta cagtgaagaa tctggctatg tttccagaca cggtacccat ggactcccag
     5281 tccctggtgg aggttagagg gtcttgtgtc aacaattcta aggaggaaga tcctccaagg
     5341 atgtactgca gtacagaagg cgaatggctt gtacccattg gcaagtgttc ctgcaatgct
     5401 ggctatgaag aaagaggttt tatgtgccaa gcttgtcgac caggtttcta caaggcattg
     5461 gatggtaata tgaagtgtgc taagtgcccg cctcacagtt ctactcagga agatggttca
     5521 atgaactgca ggtgtgagaa taattacttc cgggcagaca aagaccctcc atccatggct
     5581 tgtacccgac ctccatcttc accaagaaat gttatctcta atataaacga gacctcagtt
     5641 atcctggact ggagttggcc cctggacaca ggaggccgga aagatgttac cttcaacatc
     5701 atatgtaaaa aatgtgggtg gaatataaaa cagtgtgagc catgcagccc aaatgtccgc
     5761 ttcctccctc gacagtttgg actcaccaac accacggtga cagtgacaga ccttctggca
     5821 catactaact acacctttga gattgatgcc gttaatgggg tgtcagagct gagctcccca
     5881 ccaagacagt ttgctgcggt cagcatcaca actaatcagg ctgctccatc acctgtcctg
     5941 acgattaaga aagatcggac ctccagaaat agcatctctt tgtcctggca agaacctgaa
     6001 catcctaatg ggatcatatt ggactacgag gtcaaatact atgaaaagca ggaacaagaa
     6061 acaagttata ccattctgag ggcaagaggc acaaatgtta ccatcagtag cctcaagcct
     6121 gacactatat acgtattcca aatccgagcc cgaacagccg ctggatatgg gacgaacagc
     6181 cgcaagtttg agtttgaaac tagtccagac tctttctcca tctctggtga aagtagccaa
     6241 gtggtcatga tcgccatttc agcggcagta gcaattattc tcctcactgt tgtcatctat
     6301 gttttgattg ggaggttctg tggctataag tcaaaacatg gggcagatga aaaaagactt
     6361 cattttggca atgggcattt aaaacttcca ggtctcagga cttatgttga cccacataca
     6421 tatgaagacc ctacccaagc tgttcatgag tttgccaagg aattggatgc caccaacata
     6481 tccattgata aagttgttgg agcaggtgaa tttggagagg tgtgcagtgg tcgcttaaaa
     6541 cttccttcaa aaaaagagat ttcagtggcc attaagaccc tgaaagttgg ctacacagaa
     6601 aagcagagga gagacttcct gggagaagca agcattatgg gacagtttga ccaccccaat
     6661 atcattcgac tggaaggagt tgttaccaaa agtaagccag ttatgattgt cacagaatac
     6721 atggagaatg gttccttgga tagtttccta cgtaaacacg atgcccagtt tactgtcatt
     6781 cagctagtgg ggatgcttcg agggatagca tctggcatga agtacctgtc agacatgggc
     6841 tatgttcacc gagacctcgc tgctcggaac atcttgatca acagtaactt ggtgtgtaat
     6901 gtttctgatt tcggactttc gcgtgtcctg gaggatgacc cagaagctgc ttatacaaca
     6961 agaggaggga agatcccaat caggtggaca tcaccagaag ctatagccta ccgcaagttc
     7021 acgtcagcca gcgatgtatg gagttatggg attgttctct gggaggtgat gtcttatgga
     7081 gagagaccat actgggagat gtccaatcag gatgtaatta aagctgtaga tgagggctat
     7141 cgactgccac cccccatgga ctgcccagct gccttgtatc agctgatgct ggactgctgg
     7201 cagaaagaca ggaacaacag acccaagttt gagcagattg ttagtattct ggacaagctt
     7261 atccggaatc ccggcagcct gaagatcatc accagtgcag ccgcaaggcc atcaaacctt
     7321 cttctggacc aaagcaatgt ggatatcact accttccgca caacaggtga ctggcttaat
     7381 ggtgtctgga cagcacactg caaggaaatc ttcacgggtg tggagtacag ttcttgtgac
     7441 acaatagcca agatttccac agatgacatg aaaaaggttg gtgtcaccgt ggttgggcca
     7501 cagaagaaga tcatcagtag cattaaagct ctagagggcc cgcggttcga acaaaaactc
     7561 atctcagaag aggatctgaa tatgcatacc ggtcatcatc accatcacca ttgagtttaa
     7621 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     7681 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     7741 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     7801 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     7861 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
     7921 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     7981 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     8041 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     8101 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     8161 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     8221 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     8281 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     8341 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     8401 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     8461 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     8521 taactccgcc catcccgcc
//
