LOCUS       sequence-198290-        7113 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of c terminal EphA2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7113)
  AUTHORS   Amato KR, Wang S, Tan L, Hastings AK, Song W, Lovly CM, Meador CB, 
            Ye F, Lu P, Balko JM, Colvin DC, Cates JM, Pao W, Gray NS, Chen J
  TITLE     EPHA2 Blockade Overcomes Acquired Resistance to EGFR Kinase 
            Inhibitors in Lung Cancer.
  JOURNAL   Cancer Res. 2016 Jan 15;76(2):305-18. doi: 
            10.1158/0008-5472.CAN-15-0717. Epub 2016 Jan 7.
  PUBMED    26744526
REFERENCE   2  (bases 1 to 7113)
  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: 
            "10.1158/0008-5472.CAN-15-0717"; journalName: "Cancer Res"; date: 
            "2016-01-15- 15"; volume: "76"; issue: "2"; pages: "305-18"
COMMENT     Sequence Label: EphA2 430-976 pcDNA3.1
FEATURES             Location/Qualifiers
     source          1..7113
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     213..233
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(242..670)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(352..373)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     564..583
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    716..940
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     929..946
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(969..986)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(1002..1031)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2766..2785)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2767..2785)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2830..3033)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2859..2879)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3034..3413
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3585..3604
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3679..3783
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3784..3852,3853..4644)
                     /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(4002..4021)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4815..5403
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5304..5323
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5557..5574
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5691..5712
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5727..5732,5733..5750,5751..5757)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5765..5781
                     /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     5770..5792
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5789..5805
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5789..5805
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    5854..5975
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5865..5884)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5919..5938
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(6151..6945)
                     /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(6262..6281)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6872..6891
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(7012..7113,1..228))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(7026..7113,1..48)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7080..7099)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg gcggagttag
       61 gggcgggact atggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg
      121 ggagcctggg gactttccac acctggttgc tgactaattg agatgcatgc tttgcatact
      181 tctgcctgct ggggagcctg gggactttcc acaccctaac tgacacacat tccacagaat
      241 taattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
      301 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
      361 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
      421 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
      481 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
      541 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
      601 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
      661 cagggcgcgt ggggataccc cctagagccc cagctggttc tttccgcctc agaagccata
      721 gagcccaccg catccccagc atgcctgcta ttgtcttccc aatcctcccc cttgctgtcc
      781 tgccccaccc caccccccag aatagaatga cacctactca gacaatgcga tgcaatttcc
      841 tcattttatt aggaaaggac agtgggagtg gcaccttcca gggtcaagga aggcacgggg
      901 gaggggcaaa caacagatgg ctggcaacta gaaggcacag tcgaggctga tcagcgggtt
      961 taaactcaat ggtgatggtg atgatgaccg gtatgcatat tcagatcctc ttctgagatg
     1021 agtttttgtt cgaagggccc tctagactcg aggataggaa tccccactgt gttgacctgg
     1081 tccttgagtc ccagcaggct gtaggcaatg cgcttctggt ggccaggaag acgcactccg
     1141 atccttttga tgtcttcgtt ggacatctgt accaccttct cgatggccgt gtagccagcc
     1201 accatgaagt gttccgtgta ctgttgcatc ttgatgctct ccagccactc ggacaccgta
     1261 cggaagggga ctccctccga gccgctggtg ctgggcagcc ggatggacac tcggggatcg
     1321 aagtcagcca gcgtcttgag ggagtcgggg gctcggatga gcttgtccag gatgctaacg
     1381 atgtcggcaa acttgggccg gcgggagcgc tcttgctgcc agcactgcat catgagctgg
     1441 taaatggctg aagggcagtc catgggcgtg gggagccgga agccgtcgtt gatggctttc
     1501 atgacctcgt ggtttgacag ttcccagtag ggccgttcgc cataagtcat cacttcccac
     1561 atgacaatgc cgtagctcca cacatcgctg gctgaggtga acttgcggta ggaaatggcc
     1621 tctggggctg tccatcgaat agggatcttg ccgccacttg tggtgtaggt ggcctcgggg
     1681 tcatcctcca gcacacgcga caggccaaaa tcggacacct tgcacaccag gttgctgttg
     1741 acgaggatgt tgcgggcagc caggtcccgg tgcacgtagt tcatgttggc caggtacttc
     1801 atgccggatg cgatacccct cagcatgccc accaactgaa gtacgctgaa ctcaccatcc
     1861 ttctccctaa ggaacttgtc tagcgctcca ttctccatgt actctgtgat aatcatcatg
     1921 ggtttgtatt tagagaccac gccctccagg cggatgatat tgtggtggct aaactggccc
     1981 atgatgctgg cctcgctcag gaagtccacc cgctgcttct cagtgtagcc cgctttcagt
     2041 gtcttgatgg ccaccggtat ctccttcttc cccgaggatg ccttcagcgt ccctttatag
     2101 acctctccaa actctcctgc tccaatgacc ttctgccttg ccacacagga tgggtggatc
     2161 tcggtggtaa acttgagtac agcctggttg gggtcttcgt aagtgtgagg atccacatag
     2221 gtcttcaggg gctttagttg ttctgacttg gaaaaacgga catcctcaga ggactggcga
     2281 gcccgcaggt tcctcctcct tcgatggatg aagaggccaa ctcctgccag taccagaagc
     2341 aaaacaacac ctacagccac accgccgatc accgccatgt tggcagatcc ttccgtggac
     2401 agcgtctgga actcgtgcac tttgctgccg gctccttggc cctcctgcgt cagcgcctgc
     2461 acctgcacca ggtacgtggt atccggagca aggtcatcca gggtcacgga gaagccttcc
     2521 gtgcggcgca cattatagct gttggcatcc cccttcttgc ggtaggtgac ttcgtacttc
     2581 cacacacggc tctgctgtga caccgggatg ctccaggtga cactcaggga ggtggtgctt
     2641 cggtcctcca gcctcacttt ggggggctct gtttggttaa tactgacgct ggccatagcg
     2701 aattccacca cactggacta gtggatccga gctcggtacc aagcttaact agccagcttg
     2761 ggtctcccta tagtgagtcg tattaatttc gataagccag taagcagtgg gttctctagt
     2821 tagccagaga gctctgctta tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt
     2881 caatggggcg gagttgttac gacattttgg aaagtcccgt tgattttggt gccaaaacaa
     2941 actcccattg acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa accgctatcc
     3001 acgcccattg atgtactgcc aaaaccgcat caccatggta atagcgatga ctaatacgta
     3061 gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat gccaggcggg
     3121 ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca cttgatgtac
     3181 tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga aagtccctat
     3241 tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt cgttgggcgg
     3301 tcagccaggc gggccattta ccgtaagtta tgtaacgcgg aactccatat atgggctatg
     3361 aactaatgac cccgtaattg attactatta ataactagtc aataatcaat gtcaacgcgt
     3421 atatctggcc cgtacatcgc gaagcagcgc aaaacgccta accctaagca gattcttcat
     3481 gcaattgtcg gtcaagcctt gccttgttgt agcttaaatt ttgctcgcgc actactcagc
     3541 gacctccaac acacaagcag ggagcagata ctggcttaac tatgcggcat cagagcagat
     3601 tgtactgaga gtgcaccata ggggatcggg agatctcccg atccgtcgac gtcaggtggc
     3661 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3721 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3781 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     3841 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     3901 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     3961 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     4021 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     4081 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     4141 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     4201 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     4261 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     4321 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     4381 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     4441 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     4501 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     4561 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     4621 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     4681 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     4741 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     4801 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     4861 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     4921 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     4981 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     5041 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     5101 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     5161 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     5221 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     5281 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     5341 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     5401 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     5461 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     5521 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     5581 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     5641 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     5701 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     5761 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     5821 ctctagctag aggtcgacgg tatacagaca tgataagata cattgatgag tttggacaaa
     5881 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     5941 tatttgtaac cattataagc tgcaataaac aagttggggt gggcgaagaa ctccagcatg
     6001 agatccccgc gctggaggat catccagccg gcgtcccgga aaacgattcc gaagcccaac
     6061 ctttcataga aggcggcggt ggaatcgaaa tctcgtgatg gcaggttggg cgtcgcttgg
     6121 tcggtcattt cgcgaacccc agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa
     6181 ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc ggtcagccca
     6241 ttcgccgcca agctcttcag caatatcacg ggtagccaac gctatgtcct gatagcggtc
     6301 cgccacaccc agccggccac agtcgatgaa tccagaaaag cggccatttt ccaccatgat
     6361 attcggcaag caggcatcgc catgggtcac gacgagatcc tcgccgtcgg gcatgcgcgc
     6421 cttgagcctg gcgaacagtt cggctggcgc gagcccctga tgctcttcgt ccagatcatc
     6481 ctgatcgaca agaccggctt ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg
     6541 gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc cgccgcattg catcagccat
     6601 gatggatact ttctcggcag gagcaaggtg agatgacagg agatcctgcc ccggcacttc
     6661 gcccaatagc agccagtccc ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg
     6721 aacgcccgtc gtggccagcc acgatagccg cgctgcctcg tcctgcagtt cattcagggc
     6781 accggacagg tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca gccggaacac
     6841 ggcggcatca gagcagccga ttgtctgttg tgcccagtca tagccgaata gcctctccac
     6901 ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa acgatcctca
     6961 tcctgtctct tgatcagatc cgaaaatgga tatacaagct cccgggagct ttttgcaaaa
     7021 gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg cagaggcggc
     7081 ctcggcctct gcataaataa aaaaaattag tca
//
