LOCUS       sequence-289000-        9208 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian Expression of HsSmg6-del577-816.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9208)
  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 9208)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pCIneo-V5-SBP-HsSmg6-del577-816_I
FEATURES             Location/Qualifiers
     source          1..9208
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..729
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     680..700
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     726..750
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          890..1022
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1005..1024
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1067..1085
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1098..1107
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1107..1148
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1170..1283
                     /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"
     polyA_signal    4912..5033
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4923..4942)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4977..4996
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      5219..5674
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5306..5325)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5516..5537
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5791..6148
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5791..5982
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5814..5834)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5999..6134
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6199..6993
                     /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(6253..6272)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6863..6882
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7057..7105
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(7146..7165)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7265..7287
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7325..7343)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7411..7515
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7516..7584,7585..8376)
                     /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(7734..7753)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8547..9135
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactg
      661 cgatcgcccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      721 agcagagctc gtttagtgaa ccgtcagatc actagaagct ttattgcggt agtttatcac
      781 agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcagt
      841 gactctctta aggtagcctt gcagaagttg gtcgtgaggc actgggcagg taagtatcaa
      901 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
      961 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     1021 aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat acgactcact
     1081 ataggctagc aattgcggcc accatgggta agcctatccc taaccctctc ctcggtctcg
     1141 attctacgcc cggaagcggt agcggtacca tggacgagaa gaccaccggc tgggggggcg
     1201 gccacgtggt ggagggcctg gccggcgagc tggagcagct gcgggccagg ctggagcacc
     1261 accctcaggg ccagcgggag ccctccggcg gctgcaagcc cctcgagaat tcacgcgtga
     1321 tggcggaagg gctggagcgt gtgcggatct ccgcgtcgga gctgcgcggg atcctggcta
     1381 ctctggcccc gcaggccggg agcagagaaa acatgaagga attaaaggag gccaggccgc
     1441 gcaaagataa caggcgtcca gatctggaaa tctataagcc tggcctttct cggctaagga
     1501 acaagcccaa aatcaaggaa ccccctggga gtgaggaatt caaagatgaa attgttaatg
     1561 accgagattg ctctgctgtt gaaaatggta cacagcccgt taaagatgtc tgcaaggaac
     1621 tgaacaacca agagcagaat ggtcctatag acccagaaaa taatcgggga caagaatcct
     1681 ttcctaggac tgctggacaa gaggatcgta gtctaaaaat tatcaaaaga acaaagaaac
     1741 ccgacctgca gatctatcag cctggacgac gtttgcagac tgttagcaaa gaatccgcca
     1801 gtcgggtgga ggaggaagaa gtcctcaacc aggtagaaca actgagagta gaggaagatg
     1861 agtgtagggg aaatgttgcg aaggaggaag ttgcgaataa accagacagg gccgagatag
     1921 aaaagagccc aggtggtggg agagtagggg ctgcaaaagg agaaaaagga aagaggatgg
     1981 gaaaagggga gggggtgagg gaaacccacg acgacccggc ccgcgggagg ccgggctccg
     2041 caaagcgcta ctcccgctca gacaaacgaa ggaatcgcta ccgcacgcgc agcaccagct
     2101 cagctggcag caacaacagc gctgagggag ctggcctgac ggataatgga tgtcgccgcc
     2161 gccgacagga taggaccaag gagaggccac cactgaagaa gcaagtgtct gtgtcctcaa
     2221 ccgattcctt agacgaggac agaattgatg agcctgatgg attaggaccc aggagaagtt
     2281 cagaaaggaa gagacattta gaaagaaact ggtctggccg tggggagggt gagcagaaaa
     2341 ccagtgctaa agaatatcga ggcactcttc gtgtcacttt cgatgcagaa gccatgaaca
     2401 aagagtctcc catggtgagg tcagccaggg atgatatgga tagaggaaag cctgacaaag
     2461 gcttgagcag tgggggcaaa ggctctgaga agcaggagtc caaaaacccg aaacaagaac
     2521 ttcggggtcg tggtcgtggc attctgattt tgcctgccca taccacccta tctgtcaatt
     2581 cagcaggttc tccagagtcc gcgcctttgg gacctcggct tttgtttgga tctggtagta
     2641 agggatctcg gagttggggc cgtggaggca ccacacgccg attgtgggac ccaaacaatc
     2701 ctgatcagaa acctgctcta aagactcaga cgccccagct acatttcttg gacactgatg
     2761 atgaagtcag ccctacatct tggggtgact cacgccaggc tcaggcatct tactataagt
     2821 ttcaaaactc tgacaacccc tattattacc cccggacacc aggccctgcc tcccagtatc
     2881 cctatacggg ctataaccct ctacagtacc cagtgggccc tacgaatggt gtgtacccag
     2941 ggccttacta cccaggctac ccgactccgt caggacagta tgtgtgtagc cctctaccta
     3001 ccagcaccat gagtcccgag gaggtagagc agcacatgag gaacctgacg cgtgcagaac
     3061 agatggaaaa gaagcaacat gaggaatttg acctgagccc tgaccagtgg cggaaaggaa
     3121 agaagtctac tttccggcat gttggagatg acaccactcg cctggagatc tggattcatc
     3181 catcccatcc acggtcttcc cagggcactg agtctgggaa ggattctgag caagagaatg
     3241 ggctgggcag cctgagtccc agtgatctga acaaaaggtt catcctcagt tttctccatg
     3301 cccatgggaa gctgtttacc cggattggga tggagacatt ccctgcagtg gctgagaagg
     3361 tcctcaagga gttccaggtg ttactgcagc acagcccctc tcccattgga agtacccgca
     3421 tgctgcagct tatgaccatc aatatgtttg cagtacacaa ctcccagctg aaagactgct
     3481 tctcggagga gtgccgctct gtgatccagg aacaagccgc agctctgggc ttggccatgt
     3541 tttctctact ggtccgccgc tgcacctgct tacttaagga gtccgccaaa gctcagctgt
     3601 cctctcctga ggaccaggat gaccaagacg acatcaaggt gtcttccttt gtcccggacc
     3661 tgaaggagct gctccccagt gtcaaagtct ggtcagattg gatgctcggc tacccggaca
     3721 cctggaatcc tcctcccaca tccctggatc tgccctcgca tgttgctgtg gatgtatggt
     3781 cgacgctggc tgatttctgt aacatactga ctgcagtgaa tcagtctgag gtgccactgt
     3841 acaaggaccc ggatgatgac ctcacccttc ttatcctgga agaggatcgg cttctctcgg
     3901 gctttgtccc cttgctggct gcccctcagg acccctgcta cgtggagaaa acctcggata
     3961 aggttattgc agctgactgc aaaagggtca cagtgctgaa gtattttctg gaagcccttt
     4021 gtggacaaga agagcctctg ctggcattca agggtggaaa gtatgtgtca gtggcacccg
     4081 tcccagacac catgggaaag gaaatgggaa gccaagaggg aacacgactg gaagatgagg
     4141 aggaggatgt ggtgattgaa gactttgagg aagattcaga ggctgaaggc agcggaggcg
     4201 aggatgacat cagggagctt cgggccaaga agctggctct ggccaggaag atagctgagc
     4261 agcagcgtcg ccaggaaaag atccaggctg tcctggagga ccacagtcag atgaggcaga
     4321 tggagctcga aatcagacct ttgttcctcg taccagacac caacggcttc attgaccacc
     4381 tggccagtct ggcgcggctg ctggagagca ggaagtacat cctggtggtg cccctcatcg
     4441 tgatcaatga gctggacggc ctggccaagg ggcaggagac agaccaccgg gctgggggct
     4501 acgcccgtgt ggtacaagag aaggcccgca agtccatcga gttcctcgag cagcgattcg
     4561 agagtcggga ctcttgcctg cgagccctga ccagccgtgg caatgaactc gaatccatcg
     4621 ccttccgcag tgaggacatc actggccagc tgggtaacaa cgatgatctc atcctgtcct
     4681 gctgcctcca ctactgcaaa gacaaggcta aggacttcat gcccgccagc aaagaggagc
     4741 caatccggct actgcgggag gtggtgctgt tgacggatga ccggaacctg cgtgtgaagg
     4801 cgctcacaag gaatgttcct gtacgggaca tcccagcctt cctcacgtgg gcccaggtgg
     4861 gctgagcggc cgcttccctt tagtgagggt taatgcttcg agcagacatg ataagataca
     4921 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     4981 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     5041 acaattgcat tcattttatg tttcaggttc agggggagat gtgggaggtt ttttaaagca
     5101 agtaaaacct ctacaaatgt ggtaaaatcc gataaggatc gatccgggct ggcgtaatag
     5161 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggac
     5221 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     5281 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     5341 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     5401 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     5461 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     5521 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     5581 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     5641 gcgaatttta acaaaatatt aacgcttaca atttcctgat gcggtatttt ctccttacgc
     5701 atctgtgcgg tatttcacac cgcatacgcg gatctgcgca gcaccatggc ctgaaataac
     5761 ctctgaaaga ggaacttggt taggtacctt ctgaggcgga aagaaccagc tgtggaatgt
     5821 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     5881 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     5941 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     6001 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     6061 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     6121 cttttttgga ggcctaggct tttgcaaaaa gcttgattct tctgacacaa cagtctcgaa
     6181 cttaaggcta gagccaccat gattgaacaa gatggattgc acgcaggttc tccggccgct
     6241 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     6301 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     6361 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     6421 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     6481 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     6541 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     6601 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     6661 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     6721 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     6781 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     6841 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     6901 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     6961 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     7021 accaagcgac gcccaacctg ccatcacgat ggccgcaata aaatatcttt attttcatta
     7081 catctgtgtg ttggtttttt gtgtgaatcg atagcgataa ggatccgcgt atggtgcact
     7141 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     7201 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     7261 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     7321 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     7381 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     7441 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     7501 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     7561 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     7621 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     7681 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     7741 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     7801 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     7861 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     7921 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     7981 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     8041 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     8101 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     8161 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     8221 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     8281 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     8341 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     8401 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     8461 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     8521 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     8581 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     8641 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     8701 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     8761 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     8821 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     8881 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     8941 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     9001 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     9061 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     9121 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     9181 ccttttgctc acatggctcg acagatct
//
