LOCUS       sequence-357080-        7248 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsSmg7_633-1091.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7248)
  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 7248)
  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_633-1091_T
FEATURES             Location/Qualifiers
     source          1..7248
                     /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(2801..2819)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2802..2819)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2839..2856)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2845..3069
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3115..3543
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3202..3221)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3412..3433
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3552..3572)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3557..3886
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3737..3872
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3799..3818
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3953..4747
                     /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(4007..4026)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4617..4636
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4921..5042
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4958..4977)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5012..5031
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5091..5107)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5091..5107)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5104..5126)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5115..5131
                     /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(5139..5145,5146..5163,5164..5169))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5184..5205
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5322..5339)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5493..6081)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5573..5592)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6252..7043,7044..7112))
                     /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     6875..6894
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7113..7217)
                     /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 gggctcaggt tacaccttcc cagctggtgt ttctgtccca
     1441 ggaacctttc ttcagcctac agctcactct ccagcaggaa accaggtgca agctgggaaa
     1501 cagtcccaca ttccttacag ccagcaacgg ccctctggac cagggccaat gaaccaggga
     1561 cctcaacaat cacagccacc ttcccagcaa ccccttacat ctttaccagc tcagccaaca
     1621 gcacagtcta caagccagct gcaggttcaa gctctaactc agcaacaaca atcccctaca
     1681 aaagctgtgc cggctttggg gaaaagcccg cctcaccact ctggattcca gcagtatcaa
     1741 caggcagatg cctccaaaca gctgtggaat ccccctcagg ttcaaggccc attagggaaa
     1801 attatgcctg tgaaacagcc ctactacctt cagacccaag accccataaa actgtttgag
     1861 ccgtcattgc aacctcctgt aatgcagcag cagcctctag aaaaaaaaat gaagcctttt
     1921 cccatggagc catataacca taatccctca gaagtcaagg tcccagaatt ctactgggat
     1981 tcttcctaca gcatggctga taacagatct gtaatggcac agcaagcaaa catagaccgc
     2041 aggggcaaac ggtcaccagg aatcttccgt ccagagcagg atcctgtacc cagaatgccg
     2101 tttgagaaat ccttattgga gaagccctca gagctcatgt cacattcatc ctctttcctg
     2161 tccctcaccg gattctctct caatcaggaa agatacccaa ataatagtat gttcaatgag
     2221 gtatatggga aaaacctgac atccagctcc aaagcagaac tcagtccctc aatggccccc
     2281 caggaaacat ctctgtattc cctttttgaa gggactccgt ggtctccatc acttcctgcc
     2341 agttcagatc attcaacacc agccagccag tctcctcatt cctctaaccc aagcagccta
     2401 cccagctctc ctccaacaca caaccataat tctgttccat tctccaattt tggacccatt
     2461 gggactccag ataacaggga tagaaggact gcagatcggt ggaaaactga taagccagcc
     2521 atgggtgggt ttggcattga ttatctctca gcaacgtcat cctctgagag cagttggcat
     2581 caggccagca ctccgagtgg cacctggaca ggccatggcc cttccatgga ggattcctct
     2641 gctgtcctca tggaaagcct aaagtctatc tggtccagtt ccatgatgca tcctggacct
     2701 tctgctctgg agcagctgtt aatgcagcag aagcagaaac agcaacgggg acaaggcacc
     2761 atgaaccctc cacactgagc ggccgcctag agggccctat tctatagtgt cacctaaatg
     2821 ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     2881 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     2941 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     3001 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     3061 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     3121 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3181 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3241 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     3301 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     3361 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     3421 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     3481 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     3541 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     3601 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     3661 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     3721 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     3781 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     3841 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     3901 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     3961 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     4021 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     4081 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     4141 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     4201 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     4261 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     4321 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     4381 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     4441 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     4501 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     4561 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     4621 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     4681 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     4741 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     4801 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     4861 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     4921 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     4981 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     5041 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     5101 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     5161 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     5221 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     5281 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     5341 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     5401 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     5461 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5521 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5581 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5641 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5701 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5761 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5821 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5881 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5941 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     6001 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     6061 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     6121 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     6181 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     6241 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     6301 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     6361 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6421 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6481 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6541 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6601 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6661 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6721 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6781 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6841 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6901 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6961 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     7021 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     7081 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     7141 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     7201 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
