LOCUS       sequence-198410-        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: G766E EPHA3 pcDNA3.1
FEATURES             Location/Qualifiers
     source          1..8539
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      175..310
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     237..256
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             391..1185
                     /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(445..464)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1055..1074
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1361..1482
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1398..1417)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1452..1471
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1531..1547)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1531..1547)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1544..1566)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1555..1571
                     /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(1579..1585,1586..1603,1604..1609))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1624..1645
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1762..1779)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1933..2521)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2013..2032)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2692..3483,3484..3552))
                     /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     3315..3334
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3553..3657)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3732..3751)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3923..4302
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4303..4506
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4457..4477
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4551..4570
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4551..4569
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             7731..7760
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             7776..7793
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7816..7833)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7822..8046
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8092..8520
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8179..8198)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8389..8410
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(join(8529..8539,1..10))
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(8534..8539,1..324)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
ORIGIN
        1 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
       61 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
      121 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
      181 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
      241 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
      301 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
      361 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
      421 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
      481 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
      541 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
      601 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
      661 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
      721 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
      781 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
      841 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
      901 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
      961 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     1021 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     1081 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     1141 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     1201 tcgcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     1261 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     1321 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     1381 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     1441 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     1501 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     1561 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     1621 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     1681 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     1741 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     1801 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     1861 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     1921 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     1981 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     2041 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     2101 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     2161 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     2221 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     2281 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     2341 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     2401 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     2461 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     2521 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     2581 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     2641 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     2701 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     2761 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     2821 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     2881 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     2941 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     3001 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     3061 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     3121 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     3181 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     3241 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     3301 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     3361 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     3421 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     3481 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     3541 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     3601 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     3661 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
     3721 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
     3781 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
     3841 ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat
     3901 gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt
     3961 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     4021 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     4081 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     4141 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     4201 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     4261 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
     4321 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
     4381 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
     4441 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
     4501 agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat taatacgact
     4561 cactataggg agacccaagc tggctagtta agcttggtac cgagctcgga tccactagtc
     4621 cagtgtggtg gaattcgtcg agctctagcg aggtgacagc gtagaaccag gagtgtcaaa
     4681 cttgacatca gcctgcgagc ggagcatggt aacttctcca gcaatcagag cgctccccct
     4741 cacatcagtg gcatgcttca tggagatatg ctcctctcac tgccctctgc accagcaaca
     4801 tggattgtca gctctccatc ctcctccttc tcagctgctc tgttctcgac agcttcgggg
     4861 aactgattcc gcagccttcc aatgaagtca atctactgga ttcaaaaaca attcaagggg
     4921 agctgggctg gatctcttat ccatcacatg ggtgggaaga gatcagtggt gtggatgaac
     4981 attacacacc catcaggact taccaggtgt gcaatgtcat ggaccacagt caaaacaatt
     5041 ggctgagaac aaactgggtc cccaggaact cagctcagaa gatttatgtg gagctcaagt
     5101 tcactctacg agactgcaat agcattccat tggttttagg aacttgcaag gagacattca
     5161 acctgtacta catggagtct gatgatgatc atggggtgaa atttcgagag catcagttta
     5221 caaagattga caccattgca gctgatgaaa gtttcactca aatggatctt ggggaccgta
     5281 ttctgaagct caacactgag attagagaag taggtcctgt caacaagaag ggattttatt
     5341 tggcatttca agatgttggt gcttgtgttg ccttggtgtc tgtgagagta tacttcaaaa
     5401 agtgcccatt tacagtgaag aatctggcta tgtttccaga cacggtaccc atggactccc
     5461 agtccctggt ggaggttaga gggtcttgtg tcaacaattc taaggaggaa gatcctccaa
     5521 ggatgtactg cagtacagaa ggcgaatggc ttgtacccat tggcaagtgt tcctgcaatg
     5581 ctggctatga agaaagaggt tttatgtgcc aagcttgtcg accaggtttc tacaaggcat
     5641 tggatggtaa tatgaagtgt gctaagtgcc cgcctcacag ttctactcag gaagatggtt
     5701 caatgaactg caggtgtgag aataattact tccgggcaga caaagaccct ccatccatgg
     5761 cttgtacccg acctccatct tcaccaagaa atgttatctc taatataaac gagacctcag
     5821 ttatcctgga ctggagttgg cccctggaca caggaggccg gaaagatgtt accttcaaca
     5881 tcatatgtaa aaaatgtggg tggaatataa aacagtgtga gccatgcagc ccaaatgtcc
     5941 gcttcctccc tcgacagttt ggactcacca acaccacggt gacagtgaca gaccttctgg
     6001 cacatactaa ctacaccttt gagattgatg ccgttaatgg ggtgtcagag ctgagctccc
     6061 caccaagaca gtttgctgcg gtcagcatca caactaatca ggctgctcca tcacctgtcc
     6121 tgacgattaa gaaagatcgg acctccagaa atagcatctc tttgtcctgg caagaacctg
     6181 aacatcctaa tgggatcata ttggactacg aggtcaaata ctatgaaaag caggaacaag
     6241 aaacaagtta taccattctg agggcaagag gcacaaatgt taccatcagt agcctcaagc
     6301 ctgacactat atacgtattc caaatccgag cccgaacagc cgctggatat gggacgaaca
     6361 gccgcaagtt tgagtttgaa actagtccag actctttctc catctctggt gaaagtagcc
     6421 aagtggtcat gatcgccatt tcagcggcag tagcaattat tctcctcact gttgtcatct
     6481 atgttttgat tgggaggttc tgtggctata agtcaaaaca tggggcagat gaaaaaagac
     6541 ttcattttgg caatgggcat ttaaaacttc caggtctcag gacttatgtt gacccacata
     6601 catatgaaga ccctacccaa gctgttcatg agtttgccaa ggaattggat gccaccaaca
     6661 tatccattga taaagttgtt ggagcaggtg aatttggaga ggtgtgcagt ggtcgcttaa
     6721 aacttccttc aaaaaaagag atttcagtgg ccattaagac cctgaaagtt ggctacacag
     6781 aaaagcagag gagagacttc ctgggagaag caagcattat gggacagttt gaccacccca
     6841 atatcattcg actggaagga gttgttacca aaagtaagcc agttatgatt gtcacagaat
     6901 acatggagaa tggttccttg gatagtttcc tacgtaaaca cgatgcccag tttactgtca
     6961 ttcagctagt ggggatgctt cgagggatag catctggcat gaagtacctg tcagacatgg
     7021 gctatgttca ccgagacctc gctgctcgga acatcttgat caacagtaac ttggtgtgta
     7081 aggtttctga tttcgaactt tcgcgtgtcc tggaggatga cccagaagct gcttatacaa
     7141 caagaggagg gaagatccca atcaggtgga catcaccaga agctatagcc taccgcaagt
     7201 tcacgtcagc cagcgatgta tggagttatg ggattgttct ctgggaggtg atgtcttatg
     7261 gagagagacc atactgggag atgtccaatc aggatgtaat taaagctgta gatgagggct
     7321 atcgactgcc accccccatg gactgcccag ctgccttgta tcagctgatg ctggactgct
     7381 ggcagaaaga caggaacaac agacccaagt ttgagcagat tgttagtatt ctggacaagc
     7441 ttatccggaa tcccggcagc ctgaagatca tcaccagtgc agccgcaagg ccatcaaacc
     7501 ttcttctgga ccaaagcaat gtggatatca ctaccttccg cacaacaggt gactggctta
     7561 atggtgtctg gacagcacac tgcaaggaaa tcttcacggg tgtggagtac agttcttgtg
     7621 acacaatagc caagatttcc acagatgaca tgaaaaaggt tggtgtcacc gtggttgggc
     7681 cacagaagaa gatcatcagt agcattaaag ctctagaggg cccgcggttc gaacaaaaac
     7741 tcatctcaga agaggatctg aatatgcata ccggtcatca tcaccatcac cattgagttt
     7801 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     7861 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     7921 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     7981 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     8041 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct
     8101 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     8161 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     8221 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     8281 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     8341 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     8401 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     8461 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     8521 aattctgtgg aatgtgtgt
//
