LOCUS       sequence-198287-        6753 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of N terminal EphA2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6753)
  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 6753)
  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 1-427 pcDNA3.1
FEATURES             Location/Qualifiers
     source          1..6753
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(121..141)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        126..455
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      306..441
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     368..387
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             522..1316
                     /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(576..595)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1186..1205
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1492..1613
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1529..1548)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1583..1602
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1662..1678)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1662..1678)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1675..1697)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1686..1702
                     /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(1710..1716,1717..1734,1735..1740))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1755..1776
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1893..1910)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2064..2652)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2144..2163)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2823..3614,3615..3683))
                     /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     3446..3465
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3684..3788)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3863..3882)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4054..4433
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4434..4637
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4588..4608
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4682..4701
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4682..4700
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6076..6105
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             6121..6138
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6161..6178)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6167..6391
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(6437..6753,1..112)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6524..6543)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     join(6734..6753,1..2)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
       61 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
      121 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
      181 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
      241 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
      301 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
      361 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
      421 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
      481 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
      541 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
      601 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
      661 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
      721 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
      781 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
      841 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
      901 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
      961 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     1021 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     1081 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     1141 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     1201 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     1261 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     1321 gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     1381 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     1441 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     1501 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     1561 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     1621 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     1681 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     1741 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     1801 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     1861 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     1921 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     1981 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     2041 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     2101 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     2161 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     2221 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     2281 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     2341 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     2401 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     2461 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     2521 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     2581 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     2641 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     2701 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     2761 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     2821 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     2881 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     2941 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     3001 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     3061 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     3121 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     3181 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     3241 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     3301 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     3361 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     3421 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     3481 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     3541 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     3601 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     3661 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     3721 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     3781 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg
     3841 atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc
     3901 tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa
     3961 caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc
     4021 tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact agttattaat
     4081 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     4141 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     4201 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     4261 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     4321 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     4381 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
     4441 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
     4501 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
     4561 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
     4621 tctatataag cagagctctc tggctaacta gagaacccac tgcttactgg cttatcgaaa
     4681 ttaatacgac tcactatagg gagacccaag ctggctagtt aagcttggta ccgagctcgg
     4741 atccactagt ccagtgtggt ggaattcgct atggagctcc gggcagtcgg tttctgcctg
     4801 gcgctgctgt ggggttgcgc gctggcggcc gcggcggcac agggaaagga agttgttttg
     4861 ttggacttcg cagcaatgaa gggagagctc ggctggctca cgcaccccta tggcaaaggg
     4921 tgggacctga tgcagaacat catggacgac atgcctatct acatgtactc ggtgtgcaac
     4981 gtggtatccg gcgaccagga caactggctc cgcaccaact gggtgtaccg ggaggaggcc
     5041 gagcgcatct ttattgagct caagttcacg gtgcgagact gtaacagctt cccgggtggc
     5101 gccagctcct gcaaagagac cttcaacctc tactatgcag agtcagatgt ggactatggc
     5161 accaacttcc agaagcgcca gttcaccaag attgacacca tcgcccctga cgagatcacg
     5221 gtcagcagtg acttcgaggc tcgcaacgtc aagctgaacg tagaggagcg catggtgggg
     5281 ccccttaccc ggaagggctt ctacctggcc ttccaggaca tcggcgcctg cgtggcgctg
     5341 ctctccgttc gcgtctacta caagaagtgt cccgagatgc tgcagagctt ggctcgcttc
     5401 cccgagacca ttgctgtcgc tgtttccgat acacaacccc tggccacggt ggccggtacc
     5461 tgcgtggacc atgccgtggt gccttatggg ggcgaggggc ctctcatgca ctgcacggtg
     5521 gatggcgagt ggctggtgcc catcgggcag tgcctgtgcc aggaaggcta cgagaaggtc
     5581 gaggatgcct gccgagcctg ttctccagga ttcttcaagt ctgaggcatc tgagagccct
     5641 tgcctggagt gtccagagca taccctgcca tccacagagg gtgccacctc ctgccagtgt
     5701 gaagaaggct atttcagggc acctgaggac ccactgtcca tgtcttgcac acgtccaccc
     5761 tctgccccta actacctcac ggccattggc atgggtgcca aagtagaact gcgttggaca
     5821 gctcccaagg acactggtgg ccgccaggac attgtctaca gtgtcacttg tgaacagtgc
     5881 tggccagagt ctggcgagtg tgggccctgt gaggccagcg tgcgctattc agaacctcct
     5941 cacgccctga cccgcacgag tgtgacagtc agtgacctgg agccccacat gaactatacc
     6001 ttcgctgtcg aagcacgcaa tggcgtctca ggcctggtga ctagccgaag cttcctcgag
     6061 tctagagggc ccttcgaaca aaaactcatc tcagaagagg atctgaatat gcataccggt
     6121 catcatcacc atcaccattg agtttaaacc cgctgatcag cctcgactgt gccttctagt
     6181 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     6241 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     6301 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     6361 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     6421 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     6481 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     6541 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     6601 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     6661 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     6721 acgttcttta atagtggact cttgttccaa act
//
