LOCUS       sequence-198416-        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: A435S EPHA3 pcDNA3.1
FEATURES             Location/Qualifiers
     source          1..8539
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(16..345)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      30..165
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(84..103)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     330..350
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(359..787)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(469..490)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     681..700
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    833..1057
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1046..1063
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(1086..1103)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(1119..1148)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(4309..4328)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4310..4328)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4373..4576)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4402..4422)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4577..4956
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     5128..5147
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5222..5326
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5327..5395,5396..6187)
                     /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     complement(5545..5564)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6358..6946
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6847..6866
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7100..7117
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7234..7255
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7270..7275,7276..7293,7294..7300)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7308..7324
                     /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)."
     primer_bind     7313..7335
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7332..7348
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7332..7348
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    7397..7518
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7408..7427)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7462..7481
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(7694..8488)
                     /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(7805..7824)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     8415..8434
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 agctcccggg agctttttgc aaaagcctag gcctccaaaa aagcctcctc actacttctg
       61 gaatagctca gaggcagagg cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc
      121 atggggcgga gaatgggcgg aactgggcgg agttaggggc gggatgggcg gagttagggg
      181 cgggactatg gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga
      241 gcctggggac tttccacacc tggttgctga ctaattgaga tgcatgcttt gcatacttct
      301 gcctgctggg gagcctgggg actttccaca ccctaactga cacacattcc acagaattaa
      361 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
      421 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
      481 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
      541 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
      601 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
      661 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
      721 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
      781 ggcgcgtggg gataccccct agagccccag ctggttcttt ccgcctcaga agccatagag
      841 cccaccgcat ccccagcatg cctgctattg tcttcccaat cctccccctt gctgtcctgc
      901 cccaccccac cccccagaat agaatgacac ctactcagac aatgcgatgc aatttcctca
      961 ttttattagg aaaggacagt gggagtggca ccttccaggg tcaaggaagg cacgggggag
     1021 gggcaaacaa cagatggctg gcaactagaa ggcacagtcg aggctgatca gcgggtttaa
     1081 actcaatggt gatggtgatg atgaccggta tgcatattca gatcctcttc tgagatgagt
     1141 ttttgttcga accgcgggcc ctctagagct ttaatgctac tgatgatctt cttctgtggc
     1201 ccaaccacgg tgacaccaac ctttttcatg tcatctgtgg aaatcttggc tattgtgtca
     1261 caagaactgt actccacacc cgtgaagatt tccttgcagt gtgctgtcca gacaccatta
     1321 agccagtcac ctgttgtgcg gaaggtagtg atatccacat tgctttggtc cagaagaagg
     1381 tttgatggcc ttgcggctgc actggtgatg atcttcaggc tgccgggatt ccggataagc
     1441 ttgtccagaa tactaacaat ctgctcaaac ttgggtctgt tgttcctgtc tttctgccag
     1501 cagtccagca tcagctgata caaggcagct gggcagtcca tggggggtgg cagtcgatag
     1561 ccctcatcta cagctttaat tacatcctga ttggacatct cccagtatgg tctctctcca
     1621 taagacatca cctcccagag aacaatccca taactccata catcgctggc tgacgtgaac
     1681 ttgcggtagg ctatagcttc tggtgatgtc cacctgattg ggatcttccc tcctcttgtt
     1741 gtataagcag cttctgggtc atcctccagg acacgcgaaa gtccgaaatc agaaacctta
     1801 cacaccaagt tactgttgat caagatgttc cgagcagcga ggtctcggtg aacatagccc
     1861 atgtctgaca ggtacttcat gccagatgct atccctcgaa gcatccccac tagctgaatg
     1921 acagtaaact gggcatcgtg tttacgtagg aaactatcca aggaaccatt ctccatgtat
     1981 tctgtgacaa tcataactgg cttacttttg gtaacaactc cttccagtcg aatgatattg
     2041 gggtggtcaa actgtcccat aatgcttgct tctcccagga agtctctcct ctgcttttct
     2101 gtgtagccaa ctttcagggt cttaatggcc actgaaatct ctttttttga aggaagtttt
     2161 aagcgaccac tgcacacctc tccaaattca cctgctccaa caactttatc aatggatatg
     2221 ttggtggcat ccaattcctt ggcaaactca tgaacagctt gggtagggtc ttcatatgta
     2281 tgtgggtcaa cataagtcct gagacctgga agttttaaat gcccattgcc aaaatgaagt
     2341 cttttttcat ctgccccatg ttttgactta tagccacaga acctcccaat caaaacatag
     2401 atgacaacag tgaggagaat aattgctact gccgctgaaa tggcgatcat gaccacttgg
     2461 ctactttcac cagagatgga gaaagagtct ggactagttt caaactcaaa cttgcggctg
     2521 ttcgtcccat atccagcggc tgttcgggct cggatttgga atacgtatat agtgtcaggc
     2581 ttgaggctac tgatggtaac atttgtgcct cttgccctca gaatggtata acttgtttct
     2641 tgttcctgct tttcatagta tttgacctcg tagtccaata tgatcccatt aggatgttca
     2701 ggttcttgcc aggacaaaga gatgctattt ctggaggtcc gatctttctt aatcgtcagg
     2761 acaggtgatg gagcagactg attagttgtg atgctgaccg cagcaaactg tcttggtggg
     2821 gagctcagct ctgacacccc attaacggca tcaatctcaa aggtgtagtt agtatgtgcc
     2881 agaaggtctg tcactgtcac cgtggtgttg gtgagtccaa actgtcgagg gaggaagcgg
     2941 acatttgggc tgcatggctc acactgtttt atattccacc cacatttttt acatatgatg
     3001 ttgaaggtaa catctttccg gcctcctgtg tccaggggcc aactccagtc caggataact
     3061 gaggtctcgt ttatattaga gataacattt cttggtgaag atggaggtcg ggtacaagcc
     3121 atggatggag ggtctttgtc tgcccggaag taattattct cacacctgca gttcattgaa
     3181 ccatcttcct gagtagaact gtgaggcggg cacttagcac acttcatatt accatccaat
     3241 gccttgtaga aacctggtcg acaagcttgg cacataaaac ctctttcttc atagccagca
     3301 ttgcaggaac acttgccaat gggtacaagc cattcgcctt ctgtactgca gtacatcctt
     3361 ggaggatctt cctccttaga attgttgaca caagaccctc taacctccac cagggactgg
     3421 gagtccatgg gtaccgtgtc tggaaacata gccagattct tcactgtaaa tgggcacttt
     3481 ttgaagtata ctctcacaga caccaaggca acacaagcac caacatcttg aaatgccaaa
     3541 taaaatccct tcttgttgac aggacctact tctctaatct cagtgttgag cttcagaata
     3601 cggtccccaa gatccatttg agtgaaactt tcatcagctg caatggtgtc aatctttgta
     3661 aactgatgct ctcgaaattt caccccatga tcatcatcag actccatgta gtacaggttg
     3721 aatgtctcct tgcaagttcc taaaaccaat ggaatgctat tgcagtctcg tagagtgaac
     3781 ttgagctcca cataaatctt ctgagctgag ttcctgggga cccagtttgt tctcagccaa
     3841 ttgttttgac tgtggtccat gacattgcac acctggtaag tcctgatggg tgtgtaatgt
     3901 tcatccacac cactgatctc ttcccaccca tgtgatggat aagagatcca gcccagctcc
     3961 ccttgaattg tttttgaatc cagtagattg acttcattgg aaggctgcgg aatcagttcc
     4021 ccgaagctgt cgagaacaga gcagctgaga aggaggagga tggagagctg acaatccatg
     4081 ttgctggtgc agagggcagt gagaggagca tatctccatg aagcatgcca ctgatgtgag
     4141 ggggagcgct ctgattgctg gagaagttac catgctccgc tcgcaggctg atgtcaagtt
     4201 tgacactcct ggttctacgc tgtcacctcg ctagagctcg acgaattcca ccacactgga
     4261 ctagtggatc cgagctcggt accaagctta actagccagc ttgggtctcc ctatagtgag
     4321 tcgtattaat ttcgataagc cagtaagcag tgggttctct agttagccag agagctctgc
     4381 ttatatagac ctcccaccgt acacgcctac cgcccatttg cgtcaatggg gcggagttgt
     4441 tacgacattt tggaaagtcc cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa
     4501 tggggtggag acttggaaat ccccgtgagt caaaccgcta tccacgccca ttgatgtact
     4561 gccaaaaccg catcaccatg gtaatagcga tgactaatac gtagatgtac tgccaagtag
     4621 gaaagtccca taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga
     4681 cgtcaatagg gggcgtactt ggcatatgat acacttgatg tactgccaag tgggcagttt
     4741 accgtaaata ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa
     4801 catacgtcat tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat
     4861 ttaccgtaag ttatgtaacg cggaactcca tatatgggct atgaactaat gaccccgtaa
     4921 ttgattacta ttaataacta gtcaataatc aatgtcaacg cgtatatctg gcccgtacat
     4981 cgcgaagcag cgcaaaacgc ctaaccctaa gcagattctt catgcaattg tcggtcaagc
     5041 cttgccttgt tgtagcttaa attttgctcg cgcactactc agcgacctcc aacacacaag
     5101 cagggagcag atactggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     5161 ataggggatc gggagatctc ccgatccgtc gacgtcaggt ggcacttttc ggggaaatgt
     5221 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     5281 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     5341 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     5401 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     5461 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     5521 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     5581 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     5641 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     5701 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     5761 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     5821 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     5881 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     5941 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     6001 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     6061 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     6121 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     6181 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     6241 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     6301 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     6361 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     6421 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     6481 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     6541 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     6601 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     6661 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     6721 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     6781 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     6841 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     6901 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     6961 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     7021 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     7081 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     7141 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     7201 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
     7261 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     7321 acaatttcac acaggaaaca gctatgacca tgattacgcc aagctctagc tagaggtcga
     7381 cggtatacag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     7441 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     7501 agctgcaata aacaagttgg ggtgggcgaa gaactccagc atgagatccc cgcgctggag
     7561 gatcatccag ccggcgtccc ggaaaacgat tccgaagccc aacctttcat agaaggcggc
     7621 ggtggaatcg aaatctcgtg atggcaggtt gggcgtcgct tggtcggtca tttcgcgaac
     7681 cccagagtcc cgctcagaag aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat
     7741 cgggagcggc gataccgtaa agcacgagga agcggtcagc ccattcgccg ccaagctctt
     7801 cagcaatatc acgggtagcc aacgctatgt cctgatagcg gtccgccaca cccagccggc
     7861 cacagtcgat gaatccagaa aagcggccat tttccaccat gatattcggc aagcaggcat
     7921 cgccatgggt cacgacgaga tcctcgccgt cgggcatgcg cgccttgagc ctggcgaaca
     7981 gttcggctgg cgcgagcccc tgatgctctt cgtccagatc atcctgatcg acaagaccgg
     8041 cttccatccg agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg
     8101 tagccggatc aagcgtatgc agccgccgca ttgcatcagc catgatggat actttctcgg
     8161 caggagcaag gtgagatgac aggagatcct gccccggcac ttcgcccaat agcagccagt
     8221 cccttcccgc ttcagtgaca acgtcgagca cagctgcgca aggaacgccc gtcgtggcca
     8281 gccacgatag ccgcgctgcc tcgtcctgca gttcattcag ggcaccggac aggtcggtct
     8341 tgacaaaaag aaccgggcgc ccctgcgctg acagccggaa cacggcggca tcagagcagc
     8401 cgattgtctg ttgtgcccag tcatagccga atagcctctc cacccaagcg gccggagaac
     8461 ctgcgtgcaa tccatcttgt tcaatcatgc gaaacgatcc tcatcctgtc tcttgatcag
     8521 atccgaaaat ggatataca
//
