LOCUS       sequence-203082-        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: EPHA3_1-972 pcDNA3.1
FEATURES             Location/Qualifiers
     source          1..8539
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        168..547
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        548..751
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     702..722
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     796..815
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        796..814
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3976..4005
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4021..4038
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4061..4078)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4067..4291
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4337..4765
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4424..4443)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4634..4655
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4774..4794)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4779..5108
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4959..5094
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5021..5040
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5175..5969
                     /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(5229..5248)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5839..5858
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6145..6266
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6182..6201)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6236..6255
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6315..6331)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6315..6331)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6328..6350)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6339..6355
                     /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(6363..6369,6370..6387,6388..6393))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6408..6429
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6546..6563)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6717..7305)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6797..6816)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7476..8267,8268..8336))
                     /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     8099..8118
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8337..8441)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8516..8535)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa
       61 aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt
      121 taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac attgattatt
      181 gactagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
      241 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
      301 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
      361 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
      421 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
      481 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
      541 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
      601 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
      661 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
      721 tgtacggtgg gaggtctata taagcagagc tctctggcta actagagaac ccactgctta
      781 ctggcttatc gaaattaata cgactcacta tagggagacc caagctggct agttaagctt
      841 ggtaccgagc tcggatccac tagtccagtg tggtggaatt cgtcgagctc tagcgaggtg
      901 acagcgtaga accaggagtg tcaaacttga catcagcctg cgagcggagc atggtaactt
      961 ctccagcaat cagagcgctc cccctcacat cagtggcatg cttcatggag atatgctcct
     1021 ctcactgccc tctgcaccag caacatggat tgtcagctct ccatcctcct ccttctcagc
     1081 tgctctgttc tcgacagctt cggggaactg attccgcagc cttccaatga agtcaatcta
     1141 ctggattcaa aaacaattca aggggagctg ggctggatct cttatccatc acatgggtgg
     1201 gaagagatca gtggtgtgga tgaacattac acacccatca ggacttacca ggtgtgcaat
     1261 gtcatggacc acagtcaaaa caattggctg agaacaaact gggtccccag gaactcagct
     1321 cagaagattt atgtggagct caagttcact ctacgagact gcaatagcat tccattggtt
     1381 ttaggaactt gcaaggagac attcaacctg tactacatgg agtctgatga tgatcatggg
     1441 gtgaaatttc gagagcatca gtttacaaag attgacacca ttgcagctga tgaaagtttc
     1501 actcaaatgg atcttgggga ccgtattctg aagctcaaca ctgagattag agaagtaggt
     1561 cctgtcaaca agaagggatt ttatttggca tttcaagatg ttggtgcttg tgttgccttg
     1621 gtgtctgtga gagtatactt caaaaagtgc ccatttacag tgaagaatct ggctatgttt
     1681 ccagacacgg tacccatgga ctcccagtcc ctggtggagg ttagagggtc ttgtgtcaac
     1741 aattctaagg aggaagatcc tccaaggatg tactgcagta cagaaggcga atggcttgta
     1801 cccattggca agtgttcctg caatgctggc tatgaagaaa gaggttttat gtgccaagct
     1861 tgtcgaccag gtttctacaa ggcattggat ggtaatatga agtgtgctaa gtgcccgcct
     1921 cacagttcta ctcaggaaga tggttcaatg aactgcaggt gtgagaataa ttacttccgg
     1981 gcagacaaag accctccatc catggcttgt acccgacctc catcttcacc aagaaatgtt
     2041 atctctaata taaacgagac ctcagttatc ctggactgga gttggcccct ggacacagga
     2101 ggccggaaag atgttacctt caacatcata tgtaaaaaat gtgggtggaa tataaaacag
     2161 tgtgagccat gcagcccaaa tgtccgcttc ctccctcgac agtttggact caccaacacc
     2221 acggtgacag tgacagacct tctggcacat actaactaca cctttgagat tgatgccgtt
     2281 aatggggtgt cagagctgag ctccccacca agacagtttg ctgcggtcag catcacaact
     2341 aatcaggctg ctccatcacc tgtcctgacg attaagaaag atcggacctc cagaaatagc
     2401 atctctttgt cctggcaaga acctgaacat cctaatggga tcatattgga ctacgaggtc
     2461 aaatactatg aaaagcagga acaagaaaca agttatacca ttctgagggc aagaggcaca
     2521 aatgttacca tcagtagcct caagcctgac actatatacg tattccaaat ccgagcccga
     2581 acagccgctg gatatgggac gaacagccgc aagtttgagt ttgaaactag tccagactct
     2641 ttctccatct ctggtgaaag tagccaagtg gtcatgatcg ccatttcagc ggcagtagca
     2701 attattctcc tcactgttgt catctatgtt ttgattggga ggttctgtgg ctataagtca
     2761 aaacatgggg cagatgaaaa aagacttcat tttggcaatg ggcatttaaa acttccaggt
     2821 ctcaggactt atgttgaccc acatacatat gaagacccta cccaagctgt tcatgagttt
     2881 gccaaggaat tggatgccac caacatatcc attgataaag ttgttggagc aggtgaattt
     2941 ggagaggtgt gcagtggtcg cttaaaactt ccttcaaaaa aagagatttc agtggccatt
     3001 aagaccctga aagttggcta cacagaaaag cagaggagag acttcctggg agaagcaagc
     3061 attatgggac agtttgacca ccccaatatc attcgactgg aaggagttgt taccaaaagt
     3121 aagccagtta tgattgtcac agaatacatg gagaatggtt ccttggatag tttcctacgt
     3181 aaacacgatg cccagtttac tgtcattcag ctagtgggga tgcttcgagg gatagcatct
     3241 ggcatgaagt acctgtcaga catgggctat gttcaccgag acctcgctgc tcggaacatc
     3301 ttgatcaaca gtaacttggt gtgtaaggtt tctgatttcg gactttcgcg tgtcctggag
     3361 gatgacccag aagctgctta tacaacaaga ggagggaaga tcccaatcag gtggacatca
     3421 ccagaagcta tagcctaccg caagttcacg tcagccagcg atgtatggag ttatgggatt
     3481 gttctctggg aggtgatgtc ttatggagag agaccatact gggagatgtc caatcaggat
     3541 gtaattaaag ctgtagatga gggctatcga ctgccacccc ccatggactg cccagctgcc
     3601 ttgtatcagc tgatgctgga ctgctggcag aaagacagga acaacagacc caagtttgag
     3661 cagattgtta gtattctgga caagcttatc cggaatcccg gcagcctgaa gatcatcacc
     3721 agtgcagccg caaggccatc aaaccttctt ctggaccaaa gcaatgtgga tatcactacc
     3781 ttccgcacaa caggtgactg gcttaatggt gtctggacag cacactgcaa ggaaatcttc
     3841 acgggtgtgg agtacagttc ttgtgacaca atagccaaga tttccacaga tgacatgaaa
     3901 aaggttggtg tcaccgtggt tgggccacag aagaagatca tcagtagcat taaagctcta
     3961 gagggcccgc ggttcgaaca aaaactcatc tcagaagagg atctgaatat gcataccggt
     4021 catcatcacc atcaccattg agtttaaacc cgctgatcag cctcgactgt gccttctagt
     4081 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     4141 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     4201 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     4261 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     4321 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     4381 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     4441 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     4501 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     4561 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     4621 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     4681 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     4741 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     4801 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4861 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4921 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4981 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     5041 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     5101 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     5161 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     5221 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     5281 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     5341 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     5401 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     5461 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     5521 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     5581 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     5641 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     5701 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     5761 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     5821 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     5881 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     5941 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcga aatgaccgac
     6001 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     6061 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     6121 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     6181 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     6241 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     6301 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     6361 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     6421 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     6481 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     6541 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     6601 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     6661 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     6721 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     6781 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     6841 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     6901 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     6961 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     7021 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     7081 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     7141 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     7201 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     7261 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     7321 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     7381 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     7441 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     7501 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     7561 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     7621 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     7681 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     7741 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     7801 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     7861 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     7921 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     7981 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     8041 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     8101 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     8161 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     8221 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     8281 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     8341 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     8401 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     8461 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa
     8521 tctgctctga tgccgcata
//
