LOCUS       sequence-289926-        7976 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsSmg7-K66ER163E.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7976)
  AUTHORS   Fukuhara N, Ebert J, Unterholzner L, Lindner D, Izaurralde E, Conti 
            E
  TITLE     SMG7 is a 14-3-3-like adaptor in the nonsense-mediated mRNA decay 
            pathway.
  JOURNAL   Mol Cell. 2005 Feb 18;17(4):537-47. doi: 
            10.1016/j.molcel.2005.01.010.
  PUBMED    15721257
REFERENCE   2  (bases 1 to 7976)
  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: 
            "10.1016/j.molcel.2005.01"; journalName: "Mol Cell"; date: 
            "2005-02-18- 18"; volume: "17"; issue: "4"; pages: "537-47"
COMMENT     Sequence Label: pEYFP-N1-HsSmg7-K66ER163E_S
FEATURES             Location/Qualifiers
     source          1..7976
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3916..3925
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3922..3924,3925..3927,3928..4641)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3967..3988)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4228..4247)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4575..4596
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4764..4885
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4801..4820)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4855..4874
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4892..5347)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5029..5050)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5241..5260
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5374..5478
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5480..5837
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5480..5671
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5503..5523)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5688..5823
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5872..6666
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5872..6666
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5926..5945)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6536..6555
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6854..6873)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6898..6945
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7274..7862
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7763..7782
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagcc caacatgagc ctgcagagcg cgcagtacct ccggcaggca
      661 gaagtcctga aggctgacat gacagattct aagctgggtc cagctgaagt ctggacatcc
      721 aggcaggctc tgcaggacct gtaccagaaa atgctagtta ccgatttgga atacgcttta
      781 gacaagaaag tagaacagga tctctggaat cacgcctttg aaaatcagat cacaacactg
      841 caaggccagg caaagaatcg agcaaatccg aatcggagtg aagttcaggc aaacctttct
      901 ctgttcctag aggcagctag tggcttctat actcagttat tacaagaact gtgtacagta
      961 tttaatgtag atttaccatg ccgtgtgaag tcttcccaat tgggaattat cagcaataaa
     1021 cagacgcata ccagcgccat agtgaagcca cagtctagct cctgttccta tatctgccag
     1081 cactgcctcg tccaccttgg agacattgct gaatacagaa accagaccag ccaggcagag
     1141 tcctactata ggcatgcagc tcagcttgtc ccctccaatg gtcagcctta taatcagttg
     1201 gctatcttag cttcttccaa aggagaccat ctgaccacaa ttttctacta ctgcagaagc
     1261 attgctgtga agttcccttt cccagctgcc tccactaatc tgcaaaaagc actttctaaa
     1321 gcactggaaa gccgagatga ggtgaaaacc aagtggggtg tttctgactt catcaaggcc
     1381 tttattaaat tccacggtca tgtgtacctg agtaagagct tggaaaagtt gagccctctt
     1441 cgagagaaat tggaagaaca gtttaagagg ctgctattcc aaaaagcttt caactctcag
     1501 cagttagttc atgtcactgt cattaacctg tttcaacttc atcaccttcg tgactttagc
     1561 aatgaaaccg agcagcacac ttatagccaa gatgagcagc tatgttggac acagttgctg
     1621 gccctcttta tgtcttttct tggcatcctg tgcaagtgtc ctctacagaa tgagtctcag
     1681 gaggagtcct acaatgccta tcctcttcca gcagtcaagg tctccatgga ctggctaaga
     1741 ctcagaccca gggtctttca ggaggcagtg gtggatgaaa gacagtacat ttggccctgg
     1801 ttgatttccc ttctgaatag tttccatccc catgaagagg acctctcaag tatttaggcg
     1861 acaccacttc cagaggagtt tgaattacaa ggatttttgg cattgagacc ttctttcagg
     1921 aacttggatt tttccaaagg tcaccagggt attacagggg acaaagaagg ccagcaacga
     1981 cgaatacgac agcaacgctt gatctctata ggcaaatgga ttgctgataa tcagccaagg
     2041 ctgattcagt gtgaaaatga ggtagggaaa ttgttgttta tcacagaaat cccagaatta
     2101 atactggaag accccagtga agccaaagag aacctcattc tgcaggaaac atctgtgata
     2161 gagtcgctgg ctgcagatgg gagcccaggg ctaaaatcag tgctatctac aagccgaaat
     2221 ttaagcaaca actgtgacac aggagagaag ccagtggtta ccttcaaaga aaacattaag
     2281 acacgagaag tgaacagaga ccaaggaaga agttttcctc ccaaagaggt aaaatcccag
     2341 acagaactaa gaaagactcc agtgtctgaa gccagaaaaa cacctgtaac tcaaacccca
     2401 actcaagcaa gtaactccca gttcatcccc attcatcacc ctggagcctt ccctcctctt
     2461 cccagcaggc cagggtttcc gcccccaaca tatgttatcc ccccgcctgt ggcattttct
     2521 atgggctcag gttacacctt cccagctggt gtttctgtcc caggaacctt tcttcagcct
     2581 acagctcact ctccagcagg aaaccaggtg caagctggga aacagtccca cattccttac
     2641 agccagcaac ggccctctgg accagggcca atgaaccagg gacctcaaca atcacagcca
     2701 ccttcccagc aaccccttac atctttacca gctcagccaa cagcacagtc tacaagccag
     2761 ctgcaggttc aagctctaac tcagcaacaa caatccccta caaaagctgt gccggctttg
     2821 gggaaaagcc cgcctcacca ctctggattc cagcagtatc aacaggcaga tgcctccaaa
     2881 cagctgtgga atccccctca ggttcaaggc ccattaggga aaattatgcc tgtgaaacag
     2941 ccctactacc ttcagaccca agaccccata aaactgtttg agccgtcatt gcaacctcct
     3001 gtaatgcagc agcagcctct agaaaaaaaa atgaagcctt ttcccatgga gccatataac
     3061 cataatccct cagaagtcaa ggtcccagaa ttctactggg attcttccta cagcatggct
     3121 gataacagat ctgtaatggc acagcaagca aacatagacc gcaggggcaa acggtcacca
     3181 ggaatcttcc gtccagagca ggatcctgta cccagaatgc cgtttgagaa atccttattg
     3241 gagaagccct cagagctcat gtcacattca tcctctttcc tgtccctcac cggattctct
     3301 ctcaatcagg aaagataccc aaataatagt atgttcaatg aggtatatgg gaaaaacctg
     3361 acatccagct ccaaagcaga actcagtccc tcaatggccc cccaggaaac atctctgtat
     3421 tccctttttg aagggactcc gtggtctcca tcacttcctg ccagttcaga tcattcaaca
     3481 ccagccagcc agtctcctca ttcctctaac ccaagcagcc tacccagctc tcctccaaca
     3541 cacaaccata attctgttcc attctccaat tttggaccca ttgggactcc agataacagg
     3601 gatagaagga ctgcagatcg gtggaaaact gataagccag ccatgggtgg gtttggcatt
     3661 gattatctct cagcaacgtc atcctctgag agcagttggc atcaggccag cactccgagt
     3721 ggcacctgga caggccatgg cccttccatg gaggattcct ctgctgtcct catggaaagc
     3781 ctaaagtcta tctggtccag ttccatgatg catcctggac cttctgctct ggagcagctg
     3841 ttaatgcagc agaagcagaa acagcaacgg ggacaaggca ccatgaaccc tccacactcc
     3901 cgggatccac cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg
     3961 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     4021 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     4081 ctgcccgtgc cctggcccac cctcgtgacc accttcggct acggcctgca gtgcttcgcc
     4141 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     4201 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     4261 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     4321 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     4381 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     4441 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     4501 gtgctgctgc ccgacaacca ctacctgagc taccagtccg ccctgagcaa agaccccaac
     4561 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     4621 atggacgagc tgtacaagta aagcggccgc gactctagat cataatcagc cataccacat
     4681 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     4741 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     4801 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     4861 tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa
     4921 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     4981 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     5041 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     5101 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     5161 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     5221 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     5281 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     5341 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     5401 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     5461 tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     5521 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     5581 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     5641 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     5701 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     5761 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     5821 gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa
     5881 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     5941 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     6001 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag
     6061 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     6121 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     6181 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     6241 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     6301 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     6361 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
     6421 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     6481 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     6541 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     6601 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     6661 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     6721 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     6781 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
     6841 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     6901 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
     6961 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     7021 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     7081 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     7141 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     7201 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     7261 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     7321 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     7381 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     7441 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     7501 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     7561 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     7621 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     7681 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     7741 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     7801 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     7861 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     7921 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcat
//
