LOCUS       sequence-290004-        7767 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsSmg7_1-633.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7767)
  AUTHORS   Loh B, Jonas S, Izaurralde E
  TITLE     The SMG5-SMG7 heterodimer directly recruits the CCR4-NOT deadenylase
            complex to mRNAs containing nonsense codons via interaction with 
            POP2.
  JOURNAL   Genes Dev. 2013 Oct 1;27(19):2125-38. doi: 10.1101/gad.226951.113.
  PUBMED    24115769
REFERENCE   2  (bases 1 to 7767)
  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: 
            "10.1101/gad.226951"; journalName: "Genes Dev"; date: "2013-10-1- 
            1"; volume: "27"; issue: "19"; pages: "2125-38"
COMMENT     Sequence Label: pcDNA3.1-MS2-HA-HsSmg7_1-633_T
FEATURES             Location/Qualifiers
     source          1..7767
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      895..904
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1333..1359
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(3323..3341)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3324..3341)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3361..3378)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3367..3591
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3637..4065
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3724..3743)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3934..3955
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4074..4094)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4079..4408
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4259..4394
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4321..4340
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4475..5269
                     /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(4529..4548)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5139..5158
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5443..5564
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5480..5499)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5534..5553
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5613..5629)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5613..5629)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5626..5648)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5637..5653
                     /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(5661..5667,5668..5685,5686..5691))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5706..5727
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5844..5861)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6015..6600)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6095..6114)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6771..7562,7563..7631))
                     /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     7394..7413
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7632..7736)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttgccacc
      901 atggcttcta actttactca gttcgttctc gtcgacaatg gcggaactgg cgacgtgact
      961 gtcgccccaa gcaacttcgc taacggggtc gctgaatgga tcagctctaa ctcgcgatca
     1021 caggcttaca aagtaacctg tagcgttcgt cagagctctg cgcagaatcg caaatacacc
     1081 atcaaagtcg aggtgcctaa agtggcaacc cagactgttg gtggtgaaga gcttcctgta
     1141 gccggatgga gatcttactt aaatatggaa ctaaccattc caattttcgc cacgaattcc
     1201 gactgcgagc ttattgttaa ggcaatgcaa ggtctcctaa aagatggaaa cccgattccc
     1261 tcggccatcg cggccaactc cggcatctac ggaggtggag gtggatctgg tggatctcct
     1321 ccacatatgg cttacccata cgatgttcca gattacgctc ctccaaagct tggatccgaa
     1381 ttcccgggtc gactcgagat gagcctgcag agcgcgcagt acctccggca ggcagaagtc
     1441 ctgaaggctg acatgacaga ttctaagctg ggtccagctg aagtctggac atccaggcag
     1501 gctctgcagg acctgtacca gaaaatgcta gttaccgatt tggaatacgc tttagacaag
     1561 aaagtagaac aggatctctg gaatcacgcc tttaagaatc agatcacaac actacaaggc
     1621 caggcaaaga atcgagcaaa tccgaatcgg agtgaagttc aggcaaacct ttctctgttc
     1681 ctagaggcag ctagtggctt ctatactcag ttattacaag aactgtgtac agtatttaat
     1741 gtagatttac catgccgtgt gaagtcttcc caattgggaa ttatcagcaa taaacagacg
     1801 cataccagcg ccatagtgaa gccacagtct agctcctgtt cctatatctg ccagcactgc
     1861 ctcgtccacc ttggagacat tgctcgatac agaaaccaga ccagccaggc agagtcctac
     1921 tataggcatg cagctcagct tgtcccctcc aatggtcagc cttataatca gttggctatc
     1981 ttagcttctt ccaaaggaga ccatctgacc acaattttct actactgcag aagcattgct
     2041 gtgaagttcc ctttcccagc tgcctccact aatctgcaaa aagcactttc taaagcactg
     2101 gaaagccgag atgaggtgaa aaccaagtgg ggtgtttctg acttcatcaa ggcctttatt
     2161 aaattccacg gtcatgtgta cctgagtaag agcttggaaa agttgagccc tcttcgagag
     2221 aaattggaag aacagtttaa gaggctgcta ttccaaaaag ctttcaactc tcagcagtta
     2281 gttcatgtca ctgtcattaa cctgtttcaa cttcatcacc ttcgtgactt tagcaatgaa
     2341 accgagcagc acacttatag ccaagatgag cagctatgtt ggacacagtt gctggccctc
     2401 tttatgtctt ttcttggcat cctgtgcaag tgtcctctac agaatgagtc tcaggaggag
     2461 tcctacaatg cctatcctct tccagcagtc aaggtctcca tggactggct aagactcaga
     2521 cccagggtct ttcaggaggc agtggtggat gaaagacagt acatttggcc ctggttgatt
     2581 tctcttctga atagtttcca tccccatgaa gaggacctct caagtattag tgcgacacca
     2641 cttccagagg agtttgaatt acaaggattt ttggcattga gaccttcttt caggaacttg
     2701 gatttttcca aaggtcacca gggtattaca ggggacaaag aaggccagca acgacgaata
     2761 cgacagcaac gcttgatctc tataggcaaa tggattgctg ataatcagcc aaggctgatt
     2821 cagtgtgaaa atgaggtagg gaaattgttg tttatcacag aaatcccaga attaatactg
     2881 gaagacccca gtgaagccaa agagaacctc attctgcagg aaacatctgt gatagagtcg
     2941 ctggctgcag atgggagccc agggctaaaa tcagtgctat ctacaagccg aaatttaagc
     3001 aacaactgtg acacaggaga gaagccagtg gttaccttca aagaaaacat taagacacga
     3061 gaagtgaaca gagaccaagg aagaagtttt cctcccaaag aggtaaaatc ccagacagaa
     3121 ctaagaaaga ctccagtgtc tgaagccaga aaaacacctg taactcaaac cccaactcaa
     3181 gcaagtaact cccagttcat ccccattcat caccctggag ccttccctcc tcttcccagc
     3241 aggccagggt ttccgccccc aacatatgtt atccccccgc ctgtggcatt ttctatgtga
     3301 gcggccgcct agagggccct attctatagt gtcacctaaa tgctagagct cgctgatcag
     3361 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     3421 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     3481 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     3541 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     3601 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     3661 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     3721 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     3781 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     3841 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     3901 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     3961 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     4021 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt
     4081 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4141 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     4201 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     4261 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     4321 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
     4381 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
     4441 atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     4501 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     4561 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     4621 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
     4681 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     4741 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     4801 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     4861 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     4921 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     4981 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg
     5041 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     5101 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     5161 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     5221 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     5281 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     5341 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     5401 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
     5461 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     5521 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     5581 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     5641 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     5701 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     5761 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     5821 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     5881 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     5941 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     6001 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     6061 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     6121 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     6181 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     6241 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     6301 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     6361 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     6421 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     6481 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     6541 gctggtagcg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     6601 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     6661 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     6721 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     6781 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     6841 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     6901 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     6961 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     7021 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     7081 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     7141 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     7201 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     7261 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     7321 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     7381 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     7441 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     7501 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     7561 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     7621 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     7681 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     7741 tttccccgaa aagtgccacc tgacgtc
//
