LOCUS       sequence-198408-        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: R728L EPHA3 pcDNA3.1
FEATURES             Location/Qualifiers
     source          1..8539
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..330
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      181..316
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     243..262
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             397..1191
                     /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(451..470)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1061..1080
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1367..1488
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1404..1423)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1458..1477
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1537..1553)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1537..1553)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1550..1572)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1561..1577
                     /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(1585..1591,1592..1609,1610..1615))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1630..1651
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1768..1785)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1939..2527)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2019..2038)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2698..3489,3490..3558))
                     /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     3321..3340
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3559..3663)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3738..3757)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3929..4308
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4309..4512
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4463..4483
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4557..4576
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4557..4575
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             7737..7766
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             7782..7799
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7822..7839)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7828..8052
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8098..8526
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8185..8204)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8395..8416
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(join(8535..8539,1..16))
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
       61 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
      121 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
      181 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
      241 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
      301 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
      361 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
      421 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
      481 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
      541 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
      601 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
      661 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
      721 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
      781 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
      841 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
      901 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
      961 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     1021 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     1081 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     1141 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     1201 tggggttcgc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     1261 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     1321 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     1381 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     1441 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     1501 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     1561 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     1621 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     1681 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     1741 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     1801 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     1861 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     1921 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1981 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     2041 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     2101 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     2161 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     2221 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     2281 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     2341 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     2401 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     2461 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     2521 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     2581 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     2641 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     2701 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     2761 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     2821 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     2881 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     2941 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     3001 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     3061 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     3121 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     3181 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     3241 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     3301 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     3361 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     3421 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     3481 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     3541 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     3601 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     3661 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     3721 ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     3781 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
     3841 caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt
     3901 cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa
     3961 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     4021 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     4081 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     4141 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     4201 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     4261 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
     4321 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
     4381 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     4441 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     4501 ataagcagag ctctctggct aactagagaa cccactgctt actggcttat cgaaattaat
     4561 acgactcact atagggagac ccaagctggc tagttaagct tggtaccgag ctcggatcca
     4621 ctagtccagt gtggtggaat tcgtcgagct ctagcgaggt gacagcgtag aaccaggagt
     4681 gtcaaacttg acatcagcct gcgagcggag catggtaact tctccagcaa tcagagcgct
     4741 ccccctcaca tcagtggcat gcttcatgga gatatgctcc tctcactgcc ctctgcacca
     4801 gcaacatgga ttgtcagctc tccatcctcc tccttctcag ctgctctgtt ctcgacagct
     4861 tcggggaact gattccgcag ccttccaatg aagtcaatct actggattca aaaacaattc
     4921 aaggggagct gggctggatc tcttatccat cacatgggtg ggaagagatc agtggtgtgg
     4981 atgaacatta cacacccatc aggacttacc aggtgtgcaa tgtcatggac cacagtcaaa
     5041 acaattggct gagaacaaac tgggtcccca ggaactcagc tcagaagatt tatgtggagc
     5101 tcaagttcac tctacgagac tgcaatagca ttccattggt tttaggaact tgcaaggaga
     5161 cattcaacct gtactacatg gagtctgatg atgatcatgg ggtgaaattt cgagagcatc
     5221 agtttacaaa gattgacacc attgcagctg atgaaagttt cactcaaatg gatcttgggg
     5281 accgtattct gaagctcaac actgagatta gagaagtagg tcctgtcaac aagaagggat
     5341 tttatttggc atttcaagat gttggtgctt gtgttgcctt ggtgtctgtg agagtatact
     5401 tcaaaaagtg cccatttaca gtgaagaatc tggctatgtt tccagacacg gtacccatgg
     5461 actcccagtc cctggtggag gttagagggt cttgtgtcaa caattctaag gaggaagatc
     5521 ctccaaggat gtactgcagt acagaaggcg aatggcttgt acccattggc aagtgttcct
     5581 gcaatgctgg ctatgaagaa agaggtttta tgtgccaagc ttgtcgacca ggtttctaca
     5641 aggcattgga tggtaatatg aagtgtgcta agtgcccgcc tcacagttct actcaggaag
     5701 atggttcaat gaactgcagg tgtgagaata attacttccg ggcagacaaa gaccctccat
     5761 ccatggcttg tacccgacct ccatcttcac caagaaatgt tatctctaat ataaacgaga
     5821 cctcagttat cctggactgg agttggcccc tggacacagg aggccggaaa gatgttacct
     5881 tcaacatcat atgtaaaaaa tgtgggtgga atataaaaca gtgtgagcca tgcagcccaa
     5941 atgtccgctt cctccctcga cagtttggac tcaccaacac cacggtgaca gtgacagacc
     6001 ttctggcaca tactaactac acctttgaga ttgatgccgt taatggggtg tcagagctga
     6061 gctccccacc aagacagttt gctgcggtca gcatcacaac taatcaggct gctccatcac
     6121 ctgtcctgac gattaagaaa gatcggacct ccagaaatag catctctttg tcctggcaag
     6181 aacctgaaca tcctaatggg atcatattgg actacgaggt caaatactat gaaaagcagg
     6241 aacaagaaac aagttatacc attctgaggg caagaggcac aaatgttacc atcagtagcc
     6301 tcaagcctga cactatatac gtattccaaa tccgagcccg aacagccgct ggatatggga
     6361 cgaacagccg caagtttgag tttgaaacta gtccagactc tttctccatc tctggtgaaa
     6421 gtagccaagt ggtcatgatc gccatttcag cggcagtagc aattattctc ctcactgttg
     6481 tcatctatgt tttgattggg aggttctgtg gctataagtc aaaacatggg gcagatgaaa
     6541 aaagacttca ttttggcaat gggcatttaa aacttccagg tctcaggact tatgttgacc
     6601 cacatacata tgaagaccct acccaagctg ttcatgagtt tgccaaggaa ttggatgcca
     6661 ccaacatatc cattgataaa gttgttggag caggtgaatt tggagaggtg tgcagtggtc
     6721 gcttaaaact tccttcaaaa aaagagattt cagtggccat taagaccctg aaagttggct
     6781 acacagaaaa gcagaggaga gacttcctgg gagaagcaag cattatggga cagtttgacc
     6841 accccaatat cattcgactg gaaggagttg ttaccaaaag taagccagtt atgattgtca
     6901 cagaatacat ggagaatggt tccttggata gtttcctacg taaacacgat gcccagttta
     6961 ctgtcattca gctagtgggg atgcttctag ggatagcatc tggcatgaag tacctgtcag
     7021 acatgggcta tgttcaccga gacctcgctg ctcggaacat cttgatcaac agtaacttgg
     7081 tgtgtaaggt ttctgatttc ggactttcgc gtgtcctgga ggatgaccca gaagctgctt
     7141 atacaacaag aggagggaag atcccaatca ggtggacatc accagaagct atagcctacc
     7201 gcaagttcac gtcagccagc gatgtatgga gttatgggat tgttctctgg gaggtgatgt
     7261 cttatggaga gagaccatac tgggagatgt ccaatcagga tgtaattaaa gctgtagatg
     7321 agggctatcg actgccaccc cccatggact gcccagctgc cttgtatcag ctgatgctgg
     7381 actgctggca gaaagacagg aacaacagac ccaagtttga gcagattgtt agtattctgg
     7441 acaagcttat ccggaatccc ggcagcctga agatcatcac cagtgcagcc gcaaggccat
     7501 caaaccttct tctggaccaa agcaatgtgg atatcactac cttccgcaca acaggtgact
     7561 ggcttaatgg tgtctggaca gcacactgca aggaaatctt cacgggtgtg gagtacagtt
     7621 cttgtgacac aatagccaag atttccacag atgacatgaa aaaggttggt gtcaccgtgg
     7681 ttgggccaca gaagaagatc atcagtagca ttaaagctct agagggcccg cggttcgaac
     7741 aaaaactcat ctcagaagag gatctgaata tgcataccgg tcatcatcac catcaccatt
     7801 gagtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     7861 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     7921 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     7981 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     8041 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     8101 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     8161 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     8221 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     8281 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     8341 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     8401 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     8461 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     8521 cgaattaatt ctgtggaat
//
