LOCUS       sequence-201994-        8539 bp    DNA     circular SYN 15-APR-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 Apr 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: W250R EPHA3 pcDNA3.1
FEATURES             Location/Qualifiers
     source          1..8539
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(120..140)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        125..454
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      305..440
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     367..386
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             521..1315
                     /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(575..594)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1185..1204
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1491..1612
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1528..1547)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1582..1601
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1661..1677)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1661..1677)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1674..1696)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1685..1701
                     /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(1709..1715,1716..1733,1734..1739))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1754..1775
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1892..1909)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2063..2651)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2143..2162)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2822..3613,3614..3682))
                     /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     3445..3464
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3683..3787)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3862..3881)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4053..4432
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4433..4636
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4587..4607
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4681..4700
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4681..4699
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             7861..7890
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             7906..7923
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7946..7963)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7952..8176
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(8222..8539,1..111)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8309..8328)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     join(8519..8539,1)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
       61 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
      121 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
      181 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
      241 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
      301 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
      361 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
      421 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
      481 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
      541 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
      601 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
      661 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
      721 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
      781 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
      841 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
      901 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
      961 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     1021 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     1081 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     1141 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     1201 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     1261 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     1321 actctggggt tcgcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     1381 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     1441 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     1501 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     1561 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     1621 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     1681 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     1741 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     1801 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     1861 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     1921 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     1981 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     2041 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     2101 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     2161 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     2221 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     2281 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     2341 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     2401 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     2461 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     2521 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     2581 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     2641 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     2701 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     2761 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     2821 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     2881 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     2941 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     3001 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     3061 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     3121 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     3181 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     3241 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     3301 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     3361 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     3421 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     3481 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     3541 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     3601 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     3661 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     3721 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     3781 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga
     3841 tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct
     3901 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
     3961 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct
     4021 gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata
     4081 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     4141 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     4201 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     4261 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     4321 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     4381 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
     4441 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     4501 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     4561 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
     4621 ctatataagc agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat
     4681 taatacgact cactataggg agacccaagc tggctagtta agcttggtac cgagctcgga
     4741 tccactagtc cagtgtggtg gaattcgtcg agctctagcg aggtgacagc gtagaaccag
     4801 gagtgtcaaa cttgacatca gcctgcgagc ggagcatggt aacttctcca gcaatcagag
     4861 cgctccccct cacatcagtg gcatgcttca tggagatatg ctcctctcac tgccctctgc
     4921 accagcaaca tggattgtca gctctccatc ctcctccttc tcagctgctc tgttctcgac
     4981 agcttcgggg aactgattcc gcagccttcc aatgaagtca atctactgga ttcaaaaaca
     5041 attcaagggg agctgggctg gatctcttat ccatcacatg ggtgggaaga gatcagtggt
     5101 gtggatgaac attacacacc catcaggact taccaggtgt gcaatgtcat ggaccacagt
     5161 caaaacaatt ggctgagaac aaactgggtc cccaggaact cagctcagaa gatttatgtg
     5221 gagctcaagt tcactctacg agactgcaat agcattccat tggttttagg aacttgcaag
     5281 gagacattca acctgtacta catggagtct gatgatgatc atggggtgaa atttcgagag
     5341 catcagttta caaagattga caccattgca gctgatgaaa gtttcactca aatggatctt
     5401 ggggaccgta ttctgaagct caacactgag attagagaag taggtcctgt caacaagaag
     5461 ggattttatt tggcatttca agatgttggt gcttgtgttg ccttggtgtc tgtgagagta
     5521 tacttcaaaa agtgcccatt tacagtgaag aatctggcta tgtttccaga cacggtaccc
     5581 atggactccc agtccctggt ggaggttaga gggtcttgtg tcaacaattc taaggaggaa
     5641 gatcctccaa ggatgtactg cagtacagaa ggcgaaagac ttgtacccat tggcaagtgt
     5701 tcctgcaatg ctggctatga agaaagaggt tttatgtgcc aagcttgtcg accaggtttc
     5761 tacaaggcat tggatggtaa tatgaagtgt gctaagtgcc cgcctcacag ttctactcag
     5821 gaagatggtt caatgaactg caggtgtgag aataattact tccgggcaga caaagaccct
     5881 ccatccatgg cttgtacccg acctccatct tcaccaagaa atgttatctc taatataaac
     5941 gagacctcag ttatcctgga ctggagttgg cccctggaca caggaggccg gaaagatgtt
     6001 accttcaaca tcatatgtaa aaaatgtggg tggaatataa aacagtgtga gccatgcagc
     6061 ccaaatgtcc gcttcctccc tcgacagttt ggactcacca acaccacggt gacagtgaca
     6121 gaccttctgg cacatactaa ctacaccttt gagattgatg ccgttaatgg ggtgtcagag
     6181 ctgagctccc caccaagaca gtttgctgcg gtcagcatca caactaatca ggctgctcca
     6241 tcacctgtcc tgacgattaa gaaagatcgg acctccagaa atagcatctc tttgtcctgg
     6301 caagaacctg aacatcctaa tgggatcata ttggactacg aggtcaaata ctatgaaaag
     6361 caggaacaag aaacaagtta taccattctg agggcaagag gcacaaatgt taccatcagt
     6421 agcctcaagc ctgacactat atacgtattc caaatccgag cccgaacagc cgctggatat
     6481 gggacgaaca gccgcaagtt tgagtttgaa actagtccag actctttctc catctctggt
     6541 gaaagtagcc aagtggtcat gatcgccatt tcagcggcag tagcaattat tctcctcact
     6601 gttgtcatct atgttttgat tgggaggttc tgtggctata agtcaaaaca tggggcagat
     6661 gaaaaaagac ttcattttgg caatgggcat ttaaaacttc caggtctcag gacttatgtt
     6721 gacccacata catatgaaga ccctacccaa gctgttcatg agtttgccaa ggaattggat
     6781 gccaccaaca tatccattga taaagttgtt ggagcaggtg aatttggaga ggtgtgcagt
     6841 ggtcgcttaa aacttccttc aaaaaaagag atttcagtgg ccattaagac cctgaaagtt
     6901 ggctacacag aaaagcagag gagagacttc ctgggagaag caagcattat gggacagttt
     6961 gaccacccca atatcattcg actggaagga gttgttacca aaagtaagcc agttatgatt
     7021 gtcacagaat acatggagaa tggttccttg gatagtttcc tacgtaaaca cgatgcccag
     7081 tttactgtca ttcagctagt ggggatgctt cgagggatag catctggcat gaagtacctg
     7141 tcagacatgg gctatgttca ccgagacctc gctgctcgga acatcttgat caacagtaac
     7201 ttggtgtgta aggtttctga tttcggactt tcgcgtgtcc tggaggatga cccagaagct
     7261 gcttatacaa caagaggagg gaagatccca atcaggtgga catcaccaga agctatagcc
     7321 taccgcaagt tcacgtcagc cagcgatgta tggagttatg ggattgttct ctgggaggtg
     7381 atgtcttatg gagagagacc atactgggag atgtccaatc aggatgtaat taaagctgta
     7441 gatgagggct atcgactgcc accccccatg gactgcccag ctgccttgta tcagctgatg
     7501 ctggactgct ggcagaaaga caggaacaac agacccaagt ttgagcagat tgttagtatt
     7561 ctggacaagc ttatccggaa tcccggcagc ctgaagatca tcaccagtgc agccgcaagg
     7621 ccatcaaacc ttcttctgga ccaaagcaat gtggatatca ctaccttccg cacaacaggt
     7681 gactggctta atggtgtctg gacagcacac tgcaaggaaa tcttcacggg tgtggagtac
     7741 agttcttgtg acacaatagc caagatttcc acagatgaca tgaaaaaggt tggtgtcacc
     7801 gtggttgggc cacagaagaa gatcatcagt agcattaaag ctctagaggg cccgcggttc
     7861 gaacaaaaac tcatctcaga agaggatctg aatatgcata ccggtcatca tcaccatcac
     7921 cattgagttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     7981 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     8041 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     8101 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     8161 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     8221 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     8281 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     8341 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     8401 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     8461 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     8521 ggactcttgt tccaaactg
//
