LOCUS       sequence-289017-        6523 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsSmg6_817-1238.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6523)
  AUTHORS   Kashima I, Jonas S, Jayachandran U, Buchwald G, Conti E, Lupas AN, 
            Izaurralde E
  TITLE     SMG6 interacts with the exon junction complex via two conserved 
            EJC-binding motifs (EBMs) required for nonsense-mediated mRNA decay.
  JOURNAL   Genes Dev. 2010 Nov 1. 24(21):2440-50.
  PUBMED    20930030
REFERENCE   2  (bases 1 to 6523)
  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"; journalName: "Genes Dev. 
            2010 Nov 1. 24(21):2440-50."
COMMENT     Sequence Label: pN1-HsSmg6_817-1238-V5-SBP-MBP_I
FEATURES             Location/Qualifiers
     source          1..6523
                     /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      604..613
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1894..1935
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1957..2070
                     /codon_start=1
                     /product="streptavidin-binding peptide"
                     /label=SBP
                     /note="selected from a peptide library; binds streptavidin 
                     with nanomolar affinity (Keefe et al., 2001)"
                     /translation="MDEKTTGWGGGHVVEGLAGELEQLRARLEHHPQGQREP"
     regulatory      3052..3061
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3311..3432
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3348..3367)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3402..3421
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3439..3894)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3576..3597)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3788..3807
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3921..4025
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4027..4384
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4027..4218
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4050..4070)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4235..4370
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4419..5213
                     /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             4419..5213
                     /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(4473..4492)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5083..5102
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5401..5420)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5445..5492
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5821..6409
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6310..6329
                     /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 gctagcaatt
      601 gcggccacca tggcagaaca gatggaaaag aagcaacatg aggaatttga cctgagccct
      661 gaccagtggc ggaaaggaaa gaagtctact ttccggcatg ttggagatga caccactcgc
      721 ctggagatct ggattcatcc atcccatcca cggtcttccc agggcactga gtctgggaag
      781 gattctgagc aagagaatgg gctgggcagc ctgagtccca gtgatctgaa caaaaggttc
      841 atcctcagtt ttctccatgc ccatgggaag ctgtttaccc ggattgggat ggagacattc
      901 cctgcagtgg ctgagaaggt cctcaaggag ttccaggtgt tactgcagca cagcccctct
      961 cccattggaa gtacccgcat gctgcagctt atgaccatca atatgtttgc agtacacaac
     1021 tcccagctga aagactgctt ctcggaggag tgccgctctg tgatccagga acaagccgca
     1081 gctctgggct tggccatgtt ttctctactg gtccgccgct gcacctgctt acttaaggag
     1141 tccgccaaag ctcagctgtc ctctcctgag gaccaggatg accaagacga catcaaggtg
     1201 tcttcctttg tcccggacct gaaggagctg ctccccagtg tcaaagtctg gtcagattgg
     1261 atgctcggct acccggacac ctggaatcct cctcccacat ccctggatct gccctcgcat
     1321 gttgctgtgg atgtatggtc gacgctggct gatttctgta acatactgac tgcagtgaat
     1381 cagtctgagg tgccactgta caaggacccg gatgatgacc tcacccttct tatcctggaa
     1441 gaggatcggc ttctctcggg ctttgtcccc ttgctggctg cccctcagga cccctgctac
     1501 gtggagaaaa cctcggataa ggttattgca gctgactgca aaagggtcac agtgctgaag
     1561 tattttctgg aagccctttg tggacaagaa gagcctctgc tggcattcaa gggtggaaag
     1621 tatgtgtcag tggcacccgt cccagacacc atgggaaagg aaatgggaag ccaagaggga
     1681 acacgactgg aagatgagga ggaggatgtg gtgattgaag actttgagga agattcagag
     1741 gctgaaggca gcggaggcga ggatgacatc agggagcttc gggccaagaa gctggctctg
     1801 gccaggaaga tagctgagca gcagcgtcgc caggaaaaga tccaggctgt cctggaggac
     1861 cacagtcaga tgaggcagcc accggtccct ccaggtaagc ctatccctaa ccctctcctc
     1921 ggtctcgatt ctacgcccgg aagcggtagc ggtaccatgg acgagaagac caccggctgg
     1981 gggggcggcc acgtggtgga gggcctggcc ggcgagctgg agcagctgcg ggccaggctg
     2041 gagcaccacc ctcagggcca gcgggagccc tccggcggct gcaagcccct cgagatgaaa
     2101 atcgaagaag gtaaactggt aatctggatt aacggcgata aaggctataa cggtctcgct
     2161 gaagtcggta agaaattcga gaaagatacc ggaattaaag tcaccgttga gcatccggat
     2221 aaactggaag agaaattccc gcaggttgcg gcaactggcg atggccctga cattatcttc
     2281 tgggcacacg accgctttgg tggctacgct caatctggcc tgttggctga aatcaccccg
     2341 gacaaagcgt tccaggacaa gctgtatccg tttacctggg atgccgtacg ttacaacggc
     2401 aagctgattg cttacccgat cgctgttgaa gcgttatcgc tgatttataa caaagatctg
     2461 ctgccgaacc cgccaaaaac ctgggaagag atcccggcgc tggataaaga actgaaagcg
     2521 aaaggtaaga gcgcgctgat gttcaacctg caagaaccgt acttcacctg gccgctgatt
     2581 gctgctgacg ggggttatgc gttcaagtat gaaaacggca agtacgacat taaagacgtg
     2641 ggcgtggata acgctggcgc gaaagcgggt ctgaccttcc tggttgacct gattaaaaac
     2701 aaacacatga atgcagacac cgattactcc atcgcagaag ctgcctttaa taaaggcgaa
     2761 acagcgatga ccatcaacgg cccgtgggca tggtccaaca tcgacaccag caaagtgaat
     2821 tatggtgtaa cggtactgcc gaccttcaag ggtcaaccat ccaaaccgtt cgttggcgtg
     2881 ctgagcgcag gtattaacgc cgccagtccg aacaaagagc tggcgaaaga gttcctcgaa
     2941 aactatctgc tgactgatga aggtctggaa gcggttaata aagacaaacc gctgggtgcc
     3001 gtagcgctga agtcttacga ggaagagttg gcgaaagatc cacgtattgc cgccaccatg
     3061 gaaaacgccc agaaaggtga aatcatgccg aacatcccgc agatgtccgc tttctggtat
     3121 gccgtgcgta ctgcggtgat caacgccgcc agcggtcgtc agactgtcga tgaagccctg
     3181 aaagactaag cggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt
     3241 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     3301 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     3361 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     3421 caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     3481 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     3541 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     3601 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     3661 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     3721 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     3781 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     3841 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     3901 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3961 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     4021 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     4081 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     4141 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4201 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     4261 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     4321 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     4381 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     4441 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     4501 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     4561 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     4621 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4681 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     4741 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     4801 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     4861 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     4921 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     4981 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     5041 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     5101 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     5161 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     5221 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     5281 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     5341 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     5401 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     5461 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     5521 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     5581 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     5641 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     5701 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     5761 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5821 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5881 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     5941 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     6001 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     6061 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     6121 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     6181 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     6241 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     6301 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     6361 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     6421 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     6481 gttatcccct gattctgtgg ataaccgtat taccgccatg cat
//
