LOCUS       sequence-125786-        6326 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway ORF clone of human EGFR [NM_005228.3] with stop codon (for 
            native or N-terminal fusions).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6326)
  TITLE     Esposito lab
REFERENCE   2  (bases 1 to 6326)
  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"
COMMENT     Sequence Label: R777-E053 Hs.EGFR
FEATURES             Location/Qualifiers
     source          1..6326
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     23..45
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     37..54
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     38..54
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    84..183
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             join(186..257,258..2120,2121..2189,2190..3818)
                     /codon_start=1
                     /product="precursor of human epidermal growth factor 
                     receptor"
                     /label=EGFR
                     /translation="MRPSGTAGAALLALLAALCPASRALEEKKVCQGTSNKLTQLGTFE
                     DHFLSLQRMFNNCEVVLGNLEITYVQRNYDLSFLKTIQEVAGYVLIALNTVERIPLENL
                     QIIRGNMYYENSYALAVLSNYDANKTGLKELPMRNLQEILHGAVRFSNNPALCNVESIQ
                     WRDIVSSDFLSNMSMDFQNHLGSCQKCDPSCPNGSCWGAGEENCQKLTKIICAQQCSGR
                     CRGKSPSDCCHNQCAAGCTGPRESDCLVCRKFRDEATCKDTCPPLMLYNPTTYQMDVNP
                     EGKYSFGATCVKKCPRNYVVTDHGSCVRACGADSYEMEEDGVRKCKKCEGPCRKVCNGI
                     GIGEFKDSLSINATNIKHFKNCTSISGDLHILPVAFRGDSFTHTPPLDPQELDILKTVK
                     EITGFLLIQAWPENRTDLHAFENLEIIRGRTKQHGQFSLAVVSLNITSLGLRSLKEISD
                     GDVIISGNKNLCYANTINWKKLFGTSGQKTKIISNRGENSCKATGQVCHALCSPEGCWG
                     PEPRDCVSCRNVSRGRECVDKCNLLEGEPREFVENSECIQCHPECLPQAMNITCTGRGP
                     DNCIQCAHYIDGPHCVKTCPAGVMGENNTLVWKYADAGHVCHLCHPNCTYGCTGPGLEG
                     CPTNGPKIPSIATGMVGALLLLLVVALGIGLFMRRRHIVRKRTLRRLLQERELVEPLTP
                     SGEAPNQALLRILKETEFKKIKVLGSGAFGTVYKGLWIPEGEKVKIPVAIKELREATSP
                     KANKEILDEAYVMASVDNPHVCRLLGICLTSTVQLITQLMPFGCLLDYVREHKDNIGSQ
                     YLLNWCVQIAKGMNYLEDRRLVHRDLAARNVLVKTPQHVKITDFGLAKLLGAEEKEYHA
                     EGGKVPIKWMALESILHRIYTHQSDVWSYGVTVWELMTFGSKPYDGIPASEISSILEKG
                     ERLPQPPICTIDVYMIMVKCWMIDADSRPKFRELIIEFSKMARDPQRYLVIQGDERMHL
                     PSPTDSNFYRALMDEEDMDDVVDADEYLIPQQGFFSSPSTSRTPLLSSLSATSNNSTVA
                     CIDRNGLQSCPIKEDSFLQRYSSDPTGALTEDSIDDTFLPVPEYINQSVPKRPAGSVQN
                     PVYHNQPLNPAPSRDPHYQDPHSTAVGNPEYLNTVQPTCVNSTFDSPAHWAQKGSHQIS
                     LDNPDYQQDFFPKEAKPNGIFKGSTAENAEYLRVAPQSSEFIGA"
     protein_bind    complement(3817..3916)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             4293..5084
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      5177..5765
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5666..5685
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5919..5936
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(6095..6122)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(6214..6300)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ttaacgctag catggatgtt ttcccagtca cgacgttgta aaacgacggc cagtcttaaa
       61 attcgagctc ggtccgagga tccgaaataa tgattttatt ttgactgata gtgacctgtt
      121 cgttgcaaca aattgataag caatgctttt ttataatgcc aactttgtac aaaaaagttg
      181 gcaccatgcg accctccggg acggccgggg cagcgctcct ggcgctgctg gctgcgctct
      241 gcccggcgag tcgggctctg gaggaaaaga aagtttgcca aggcacgagt aacaagctca
      301 cgcagttggg cacttttgaa gatcattttc tcagcctcca gaggatgttc aataactgtg
      361 aggtggtcct tgggaatttg gaaattacct atgtgcagag gaattatgat ctttccttct
      421 taaagaccat ccaggaggtg gctggttatg tcctcattgc cctcaacaca gtggagcgaa
      481 ttcctttgga aaacctgcag atcatcagag gaaatatgta ctacgaaaat tcctatgcct
      541 tagcagtctt atctaactat gatgcaaata aaaccggact gaaggagctg cccatgagaa
      601 atttacagga aatcctgcat ggcgccgtgc ggttcagcaa caaccctgcc ctgtgcaacg
      661 tggagagcat ccagtggcgg gacatagtca gcagtgactt tctcagcaac atgtcgatgg
      721 acttccagaa ccacctgggc agctgccaaa agtgtgatcc aagctgtccc aatgggagct
      781 gctggggtgc aggagaggag aactgccaga aactgaccaa aatcatctgt gcccagcagt
      841 gctccgggcg ctgccgtggc aagtccccca gtgactgctg ccacaaccag tgtgctgcag
      901 gctgcacagg cccccgggag agcgactgcc tggtctgccg caaattccga gacgaagcca
      961 cgtgcaagga cacctgcccc ccactcatgc tctacaaccc caccacgtac cagatggatg
     1021 tgaaccccga gggcaaatac agctttggtg ccacctgcgt gaagaagtgt ccccgtaatt
     1081 atgtggtgac agatcacggc tcgtgcgtcc gagcctgtgg ggccgacagc tatgagatgg
     1141 aggaagacgg cgtacgtaag tgtaagaagt gcgaagggcc ttgccgcaaa gtgtgtaacg
     1201 gaataggtat tggtgaattt aaagactcac tctccataaa tgctacgaat attaaacact
     1261 tcaaaaactg cacctccatc agtggcgatc tccacatcct gccggtggca tttaggggtg
     1321 actccttcac acatactcct cctctggatc cacaggaact ggatattctg aaaaccgtaa
     1381 aggaaatcac agggtttttg ctgattcagg cttggcctga aaacaggacg gacctccatg
     1441 cctttgagaa cctagaaatc atacgcggca ggaccaagca acatggtcag ttttctcttg
     1501 cagtcgtcag cctgaacata acatccttgg gattacgctc cctcaaggag ataagtgatg
     1561 gagatgtgat aatttcagga aacaaaaatt tgtgctatgc aaatacaata aactggaaaa
     1621 aactgtttgg gacctccggt cagaaaacca aaattataag caacagaggt gaaaacagct
     1681 gcaaggccac aggccaggtc tgccacgcgt tgtgctcccc cgagggctgc tggggcccgg
     1741 agcccaggga ctgcgtctct tgccggaatg tcagccgagg cagggaatgc gtggacaagt
     1801 gcaaccttct ggagggtgag ccaagggagt ttgtggagaa ctctgagtgc atacagtgcc
     1861 acccagagtg cctgcctcag gccatgaaca tcacctgcac aggacgggga ccagacaact
     1921 gtatccagtg tgcccactac attgacggcc cccactgcgt caagacctgc ccggcaggag
     1981 tcatgggaga aaacaacacc ctggtctgga agtacgcaga cgccggccat gtgtgccacc
     2041 tgtgccatcc aaactgcacc tacggatgca ctgggccagg tcttgaaggc tgtccaacga
     2101 atgggcctaa gatcccgtcc atcgccactg ggatggtggg ggccctcctc ttgctgctgg
     2161 tggtggccct ggggatcggc ctcttcatgc gaaggcgcca catcgttcgg aagcgcacgc
     2221 tgcggaggct gctgcaggag agggagcttg tggagcctct tacacccagt ggagaagctc
     2281 ccaaccaagc tctcttgagg atcttgaagg aaactgaatt caaaaagatc aaagtgctgg
     2341 gctccggtgc gttcggcacg gtgtataagg gactctggat cccagaaggt gagaaagtta
     2401 aaattcccgt cgctatcaag gaattaagag aagcaacatc tccgaaagcc aacaaggaaa
     2461 tcctcgatga agcctacgtg atggccagcg tggacaaccc ccacgtgtgc cgcctgctgg
     2521 gcatctgcct cacctccacc gtgcaactca tcacgcagct catgcccttc ggctgcctcc
     2581 tggactatgt ccgggaacac aaagacaata ttggctccca gtacctgctc aactggtgtg
     2641 tgcagatcgc aaagggcatg aactacttgg aggaccgtcg cttggtgcac cgcgacctgg
     2701 cagccaggaa cgtactggtg aaaacaccgc agcatgtcaa gatcacagat tttgggctgg
     2761 ccaaactgct gggtgcggaa gagaaagaat accatgcaga aggaggcaaa gtgcctatca
     2821 agtggatggc attggaatca attttacaca gaatctatac ccaccagagt gatgtctgga
     2881 gctacggggt gaccgtttgg gagttgatga cctttggatc caagccatat gacggaatcc
     2941 ctgccagcga gatctcctcc atcctggaga aaggagaacg cctccctcag ccacccatat
     3001 gtaccatcga tgtctacatg atcatggtca agtgctggat gatagacgca gatagtcgcc
     3061 caaagttccg tgagttgatc atcgaattct ccaaaatggc ccgagacccc cagcgctacc
     3121 ttgtcattca gggggatgaa agaatgcatt tgccaagtcc tacagactcc aacttctacc
     3181 gtgccctgat ggatgaagaa gacatggacg acgtggtgga tgccgacgag tacctcatcc
     3241 cacagcaggg cttcttcagc agcccctcca cgtcacggac tcccctcctg agctctctga
     3301 gtgcaaccag caacaattcc accgtggctt gcattgatag aaatgggctg caaagctgtc
     3361 ccatcaagga agacagcttc ttgcagcgat acagctcaga ccccacaggc gccttgactg
     3421 aggacagcat agacgacacc ttcctcccag tgcctgaata cataaaccag tccgttccca
     3481 aaaggcccgc tggctctgtg cagaatcctg tctatcacaa tcagcctctg aaccccgcgc
     3541 ccagcagaga cccacactac caggaccccc acagcactgc agtgggcaac cccgagtatc
     3601 tcaacactgt ccagcccacc tgtgtcaaca gcacattcga cagccctgcc cactgggccc
     3661 agaaaggcag ccaccaaatt agcctggaca accctgacta ccagcaggac ttctttccca
     3721 aggaagccaa gccaaatggc atctttaagg gctccacagc tgaaaatgca gaatacctaa
     3781 gggtcgcgcc acaaagcagt gaatttattg gagcataatc aactttcttg tacaaagttg
     3841 gcattatagg aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa
     3901 ataaaatcat tatttgcatg caagcttaga tcctctagac cagccaggac agaaatgcct
     3961 cgacttcgct gctacccaag gttgccgggt gacgcacacc gtggaaacgg atgaaggcac
     4021 gaacccagtg gacataagcc tgttcggttc gtaagctgta atgcaagtag cgtatgcgct
     4081 cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg taacggcgca gtggcggttt
     4141 tcatggcttg ttatgactgt ttttttgggg tacagtctat gcctcgggca tccaagcagc
     4201 aagcgcgtta cgccgtgggt cgatgtttga tgttatggag cagcaacgat gttacgcagc
     4261 agggcagtcg ccctaaaaca aagttaaaca ttatgaggga agcggtgatc gccgaagtat
     4321 cgactcaact atcagaggta gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg
     4381 ccgtacattt gtacggctcc gcagtggatg gcggcctgaa gccacacagt gatattgatt
     4441 tgctggttac ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg atcaacgacc
     4501 ttttggaaac ttcggcttcc cctggagaga gcgagattct ccgcgctgta gaagtcacca
     4561 ttgttgtgca cgacgacatc attccgtggc gttatccagc taagcgcgaa ctgcaatttg
     4621 gagaatggca gcgcaatgac attcttgcag gtatcttcga gccagccacg atcgacattg
     4681 atctggctat cttgctgaca aaagcaagag aacatagcgt tgccttggta ggtccagcgg
     4741 cggaggaact ctttgatccg gttcctgaac aggatctatt tgaggcgcta aatgaaacct
     4801 taacgctatg gaactcgccg cccgactggg ctggcgatga gcgaaatgta gtgcttacgt
     4861 tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat gtcgctgccg
     4921 actgggcaat ggagcgcctg ccggcccagt atcagcccgt catacttgaa gctagacagg
     4981 cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg gaagaatttg
     5041 tccactacgt gaaaggcgag atcaccaagg tagtcggcaa ataaccctcg agccacccat
     5101 gaccaaaatc ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa
     5161 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     5221 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     5281 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     5341 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     5401 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5461 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     5521 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcat tgagaaagcg
     5581 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     5641 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     5701 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     5761 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     5821 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     5881 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     5941 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     6001 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatacgc
     6061 gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt
     6121 ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc ctccgggccg
     6181 ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc ctactcagga gagcgttcac
     6241 cgacaaacaa cagataaaac gaaaggccca gtcttccgac tgagcctttc gttttatttg
     6301 atgcctggca gttccctact ctcgcg
//
