LOCUS       sequence-198413-        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: N379K EPHA3 pcDNA3.1
FEATURES             Location/Qualifiers
     source          1..8539
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      41..176
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     103..122
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             257..1051
                     /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(311..330)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     921..940
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1227..1348
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1264..1283)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1318..1337
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1397..1413)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1397..1413)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1410..1432)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1421..1437
                     /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(1445..1451,1452..1469,1470..1475))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1490..1511
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1628..1645)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1799..2387)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1879..1898)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2558..3349,3350..3418))
                     /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     3181..3200
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3419..3523)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3598..3617)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3789..4168
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4169..4372
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4323..4343
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4417..4436
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4417..4435
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             7597..7626
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             7642..7659
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7682..7699)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7688..7912
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7958..8386
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8045..8064)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8255..8276
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8395..8415)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(8400..8539,1..190)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
ORIGIN
        1 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
       61 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
      121 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
      181 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg
      241 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
      301 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
      361 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
      421 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
      481 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
      541 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
      601 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
      661 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
      721 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
      781 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
      841 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
      901 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
      961 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     1021 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcgc gaaatgaccg
     1081 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     1141 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     1201 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     1261 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     1321 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     1381 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     1441 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     1501 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     1561 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     1621 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     1681 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     1741 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     1801 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     1861 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     1921 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     1981 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     2041 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2101 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2161 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2221 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2281 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     2341 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2401 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     2461 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     2521 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     2581 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     2641 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     2701 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     2761 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     2821 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     2881 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     2941 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     3001 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     3061 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     3121 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     3181 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     3241 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     3301 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     3361 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     3421 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     3481 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     3541 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac
     3601 aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt
     3661 cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg
     3721 catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata
     3781 tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
     3841 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     3901 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     3961 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     4021 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     4081 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     4141 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
     4201 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
     4261 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
     4321 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctctctggct
     4381 aactagagaa cccactgctt actggcttat cgaaattaat acgactcact atagggagac
     4441 ccaagctggc tagttaagct tggtaccgag ctcggatcca ctagtccagt gtggtggaat
     4501 tcgtcgagct ctagcgaggt gacagcgtag aaccaggagt gtcaaacttg acatcagcct
     4561 gcgagcggag catggtaact tctccagcaa tcagagcgct ccccctcaca tcagtggcat
     4621 gcttcatgga gatatgctcc tctcactgcc ctctgcacca gcaacatgga ttgtcagctc
     4681 tccatcctcc tccttctcag ctgctctgtt ctcgacagct tcggggaact gattccgcag
     4741 ccttccaatg aagtcaatct actggattca aaaacaattc aaggggagct gggctggatc
     4801 tcttatccat cacatgggtg ggaagagatc agtggtgtgg atgaacatta cacacccatc
     4861 aggacttacc aggtgtgcaa tgtcatggac cacagtcaaa acaattggct gagaacaaac
     4921 tgggtcccca ggaactcagc tcagaagatt tatgtggagc tcaagttcac tctacgagac
     4981 tgcaatagca ttccattggt tttaggaact tgcaaggaga cattcaacct gtactacatg
     5041 gagtctgatg atgatcatgg ggtgaaattt cgagagcatc agtttacaaa gattgacacc
     5101 attgcagctg atgaaagttt cactcaaatg gatcttgggg accgtattct gaagctcaac
     5161 actgagatta gagaagtagg tcctgtcaac aagaagggat tttatttggc atttcaagat
     5221 gttggtgctt gtgttgcctt ggtgtctgtg agagtatact tcaaaaagtg cccatttaca
     5281 gtgaagaatc tggctatgtt tccagacacg gtacccatgg actcccagtc cctggtggag
     5341 gttagagggt cttgtgtcaa caattctaag gaggaagatc ctccaaggat gtactgcagt
     5401 acagaaggcg aatggcttgt acccattggc aagtgttcct gcaatgctgg ctatgaagaa
     5461 agaggtttta tgtgccaagc ttgtcgacca ggtttctaca aggcattgga tggtaatatg
     5521 aagtgtgcta agtgcccgcc tcacagttct actcaggaag atggttcaat gaactgcagg
     5581 tgtgagaata attacttccg ggcagacaaa gaccctccat ccatggcttg tacccgacct
     5641 ccatcttcac caagaaatgt tatctctaat ataaacgaga cctcagttat cctggactgg
     5701 agttggcccc tggacacagg aggccggaaa gatgttacct tcaacatcat atgtaaaaaa
     5761 tgtgggtgga atataaaaca gtgtgagcca tgcagcccaa aggtccgctt cctccctcga
     5821 cagtttggac tcaccaacac cacggtgaca gtgacagacc ttctggcaca tactaactac
     5881 acctttgaga ttgatgccgt taatggggtg tcagagctga gctccccacc aagacagttt
     5941 gctgcggtca gcatcacaac taatcaggct gctccatcac ctgtcctgac gattaagaaa
     6001 gatcggacct ccagaaatag catctctttg tcctggcaag aacctgaaca tcctaatggg
     6061 atcatattgg actacgaggt caaatactat gaaaagcagg aacaagaaac aagttatacc
     6121 attctgaggg caagaggcac aaatgttacc atcagtagcc tcaagcctga cactatatac
     6181 gtattccaaa tccgagcccg aacagccgct ggatatggga cgaacagccg caagtttgag
     6241 tttgaaacta gtccagactc tttctccatc tctggtgaaa gtagccaagt ggtcatgatc
     6301 gccatttcag cggcagtagc aattattctc ctcactgttg tcatctatgt tttgattggg
     6361 aggttctgtg gctataagtc aaaacatggg gcagatgaaa aaagacttca ttttggcaat
     6421 gggcatttaa aacttccagg tctcaggact tatgttgacc cacatacata tgaagaccct
     6481 acccaagctg ttcatgagtt tgccaaggaa ttggatgcca ccaacatatc cattgataaa
     6541 gttgttggag caggtgaatt tggagaggtg tgcagtggtc gcttaaaact tccttcaaaa
     6601 aaagagattt cagtggccat taagaccctg aaagttggct acacagaaaa gcagaggaga
     6661 gacttcctgg gagaagcaag cattatggga cagtttgacc accccaatat cattcgactg
     6721 gaaggagttg ttaccaaaag taagccagtt atgattgtca cagaatacat ggagaatggt
     6781 tccttggata gtttcctacg taaacacgat gcccagttta ctgtcattca gctagtgggg
     6841 atgcttcgag ggatagcatc tggcatgaag tacctgtcag acatgggcta tgttcaccga
     6901 gacctcgctg ctcggaacat cttgatcaac agtaacttgg tgtgtaaggt ttctgatttc
     6961 ggactttcgc gtgtcctgga ggatgaccca gaagctgctt atacaacaag aggagggaag
     7021 atcccaatca ggtggacatc accagaagct atagcctacc gcaagttcac gtcagccagc
     7081 gatgtatgga gttatgggat tgttctctgg gaggtgatgt cttatggaga gagaccatac
     7141 tgggagatgt ccaatcagga tgtaattaaa gctgtagatg agggctatcg actgccaccc
     7201 cccatggact gcccagctgc cttgtatcag ctgatgctgg actgctggca gaaagacagg
     7261 aacaacagac ccaagtttga gcagattgtt agtattctgg acaagcttat ccggaatccc
     7321 ggcagcctga agatcatcac cagtgcagcc gcaaggccat caaaccttct tctggaccaa
     7381 agcaatgtgg atatcactac cttccgcaca acaggtgact ggcttaatgg tgtctggaca
     7441 gcacactgca aggaaatctt cacgggtgtg gagtacagtt cttgtgacac aatagccaag
     7501 atttccacag atgacatgaa aaaggttggt gtcaccgtgg ttgggccaca gaagaagatc
     7561 atcagtagca ttaaagctct agagggcccg cggttcgaac aaaaactcat ctcagaagag
     7621 gatctgaata tgcataccgg tcatcatcac catcaccatt gagtttaaac ccgctgatca
     7681 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     7741 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     7801 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     7861 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     7921 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     7981 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     8041 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     8101 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     8161 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     8221 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     8281 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     8341 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     8401 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     8461 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     8521 gtatgcaaag catgcatct
//
