LOCUS       sequence-290092-        7485 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian Expression of HsSmg7_1-633-shRNAres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7485)
  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 7485)
  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"; doi: 
            "10.1101/gad.226951"; journalName: "Genes Dev"; date: "2013-10-1- 
            1"; volume: "27"; issue: "19"; pages: "2125-38"
COMMENT     Sequence Label: pCIneo-lambdaN-HA-HsSmg7_1-633-shRNAres_U
FEATURES             Location/Qualifiers
     source          1..7485
                     /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             1104..1169
                     /codon_start=1
                     /product="N-terminal RNA-binding domain from the lambda 
                     bacteriophage antiterminator protein N (Baron-Benhamou et 
                     al., 2004)"
                     /label=lambda N peptide
                     /translation="MDAQTRRRERRAEKQAQWKAAN"
     CDS             1194..1220
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    3189..3310
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3200..3219)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3254..3273
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3496..3951
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3583..3602)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3793..3814
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4068..4425
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4068..4259
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4091..4111)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4276..4411
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4476..5270
                     /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(4530..4549)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5140..5159
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5334..5382
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5423..5442)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5542..5564
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5602..5620)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5688..5792
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5793..5861,5862..6653)
                     /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(6011..6030)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6824..7412
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7313..7332
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
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 accatggacg cacaaacacg acgacgtgag cgtcgcgctg
     1141 agaaacaagc tcaatggaaa gctgcaaacg gtggatctcc tccacatatg gcttacccat
     1201 acgatgttcc agattacgct cctccactcg agatgagcct gcagagcgcg cagtacctcc
     1261 ggcaggcaga agtcctgaag gctgacatga cagattctaa gctgggtcca gctgaagtct
     1321 ggacatccag gcaggctctg caggacctgt accagaaaat gctagttacc gatttggaat
     1381 acgctttaga caagaaagta gaacaggatc tctggaatca cgcctttaag aatcagatca
     1441 caacactaca aggccaggca aagaatcgag caaatccgaa tcggagtgaa gttcaggcaa
     1501 acctttctct gttcctagag gcagctagtg gcttctatac tcagttatta caagaactgt
     1561 gtacagtatt taatgtagat ttaccatgcc gtgtgaagtc ttcccaattg ggaattatca
     1621 gcaataaaca gacgcatacc agcgccatag tgaagccaca gtctagctcc tgttcctata
     1681 tctgccagca ctgcctcgtc caccttggag acattgctcg atacagaaac cagaccagcc
     1741 aggcagagtc ctactatagg catgcagctc agcttgtccc ctccaatggt cagccttata
     1801 atcagttggc tatcttagct tcttccaaag gagaccatct gaccacaatt ttctactact
     1861 gcagaagcat tgctgtgaag ttccctttcc cagctgcctc cactaatctg caaaaagcac
     1921 tttctaaagc actggaaagc cgagatgagg tgaaaaccaa gtggggtgtt tctgacttca
     1981 tcaaggcctt tattaaattc cacggtcatg tgtacctgag taagagcttg gaaaagttga
     2041 gccctcttcg agagaaattg gaagaacagt ttaagaggct gctattccaa aaagctttca
     2101 actctcagca gttagttcat gtcactgtca ttaacctgtt tcaacttcat caccttcgtg
     2161 actttagcaa tgaaaccgag cagcacactt atagccaaga tgagcagcta tgttggacac
     2221 agttgctggc cctctttatg tcttttcttg gcatcctgtg caagtgtcct ctacagaatg
     2281 agtctcagga ggagtcctac aatgcctatc ctcttccagc agtcaaggtc tccatggact
     2341 ggctaagact cagacccagg gtctttcagg aggcagtggt ggatgaaaga cagtacattt
     2401 ggccctggtt gatttccctt ctgaatagtt tccatcccca tgaagaggac ctctcaagca
     2461 tctccgcaac tccgctacca gaggagtttg aattacaagg atttttggca ttgagacctt
     2521 ctttcaggaa cttggatttt tccaaaggtc accagggtat tacaggggac aaagaaggcc
     2581 agcaacgacg aatacgacag caacgcttga tctctatagg caaatggatt gctgataatc
     2641 agccaaggct gattcagtgt gaaaatgagg tagggaaatt gttgtttatc acagaaatcc
     2701 cagaattaat actggaagac cccagtgaag ccaaagagaa cctcattctg caggaaacat
     2761 ctgtgataga gtcgctggct gcagatggga gcccagggct aaaatcagtg ctatctacaa
     2821 gccgaaattt aagcaacaac tgtgacacag gagagaagcc agtggttacc ttcaaagaaa
     2881 acattaagac acgagaagtg aacagagacc aaggaagaag ttttcctccc aaagaggtaa
     2941 aatcccagac agaactaaga aagactccag tgtctgaagc cagaaaaaca cctgtaactc
     3001 aaaccccaac tcaagcaagt aactcccagt tcatccccat tcatcaccct ggagccttcc
     3061 ctcctcttcc cagcaggcca gggtttccgc ccccaacata tgttatcccc ccgcctgtgg
     3121 cattttctat gtcccgggat ccgcggccgc ttccctttag tgagggttaa tgcttcgagc
     3181 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     3241 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     3301 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggagatgtg
     3361 ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatccgat aaggatcgat
     3421 ccgggctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     3481 ctgaatggcg aatggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     3541 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     3601 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     3661 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     3721 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     3781 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     3841 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     3901 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt tcctgatgcg
     3961 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgcggat ctgcgcagca
     4021 ccatggcctg aaataacctc tgaaagagga acttggttag gtaccttctg aggcggaaag
     4081 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     4141 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     4201 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4261 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4321 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     4381 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct tgattcttct
     4441 gacacaacag tctcgaactt aaggctagag ccaccatgat tgaacaagat ggattgcacg
     4501 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     4561 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     4621 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     4681 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     4741 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     4801 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     4861 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     4921 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     4981 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     5041 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     5101 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     5161 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     5221 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     5281 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgatggc cgcaataaaa
     5341 tatctttatt ttcattacat ctgtgtgttg gttttttgtg tgaatcgata gcgataagga
     5401 tccgcgtatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     5461 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     5521 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     5581 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     5641 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     5701 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     5761 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     5821 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     5881 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     5941 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     6001 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     6061 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     6121 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     6181 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     6241 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     6301 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     6361 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     6421 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     6481 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     6541 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     6601 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     6661 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     6721 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     6781 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     6841 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     6901 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     6961 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     7021 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     7081 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     7141 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     7201 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     7261 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     7321 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     7381 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     7441 tcctggcctt ttgctggcct tttgctcaca tggctcgaca gatct
//
