LOCUS       sequence-288979-        8152 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsSmg7_236-1091-siRNAres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8152)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 8152)
  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"
COMMENT     Sequence Label: pCIneo-lambdaN-HA-HsSmg7_236-1091-siRNAres_I
FEATURES             Location/Qualifiers
     source          1..8152
                     /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    3856..3977
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3867..3886)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3921..3940
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      4163..4618
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4250..4269)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4460..4481
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4735..5092
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4735..4926
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4758..4778)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4943..5078
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5143..5937
                     /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(5197..5216)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5807..5826
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6001..6049
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(6090..6109)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6209..6231
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6269..6287)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6355..6459
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6460..6528,6529..7320)
                     /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(6678..6697)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7491..8079
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7980..7999
                     /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 agacgcgtag ccgagatgag gtgaaaacca
     1261 agtggggtgt ttctgacttc atcaaggcct ttattaaatt ccacggtcat gtgtacctga
     1321 gtaagagctt ggaaaagttg agccctcttc gagagaaatt ggaagaacag tttaagaggc
     1381 tgctattcca aaaagctttc aactctcagc agttagttca tgtcactgtc attaacctgt
     1441 ttcaacttca tcaccttcgt gactttagca atgaaaccga gcagcacact tatagccaag
     1501 atgagcagct atgttggaca cagttgctgg ccctctttat gtcttttctt ggcatcctgt
     1561 gcaagtgtcc tctacagaat gagtctcagg aggagtccta caatgcctat cctcttccag
     1621 cagtcaaggt ctccatggac tggctaagac tcagacccag ggtctttcag gaggcagtgg
     1681 tggatgaaag acagtacatt tggccctggt tgatttccct tctgaatagt ttccatcccc
     1741 atgaagagga cctctcaagt attagtgcga caccacttcc agaggagttt gaattacaag
     1801 gatttttggc attgagacct tctttcagga acttggattt ttccaaaggt caccagggta
     1861 ttacagggga caaagaaggc cagcaacgac gaatacgaca gcaacgcttg atctctatag
     1921 gcaaatggat tgctgataat cagccaaggc tgattcagtg tgaaaatgag gtagggaaat
     1981 tgttgtttat cacagaaatc ccagaattaa tactggaaga ccccagtgaa gccaaagaga
     2041 acctcattct gcaggaaaca tctgtgatag agtcgctggc tgcagatggg agcccagggc
     2101 taaaatcagt gctatctaca agccgaaatt taagcaacaa ctgtgacaca ggagagaagc
     2161 cagtggttac cttcaaagaa aacattaaga cacgagaagt gaacagagac caaggaagaa
     2221 gttttcctcc caaagaggta aaatcccaga cagaactaag aaagactcca gtgtctgaag
     2281 ccagaaaaac acctgtaact caaaccccaa ctcaagcaag taactcccag ttcatcccca
     2341 ttcatcaccc tggagccttc cctcctcttc ccagcaggcc agggtttccg cccccaacat
     2401 atgttatccc cccgcctgtg gcattttcta tgggctcagg ttacaccttc ccagctggtg
     2461 tttctgtccc aggaaccttt cttcggccta cagctcactc tccagcagga aaccaggtgc
     2521 aagctgggaa acagtcccac attccttaca gccagcaacg gccctctgga ccagggccaa
     2581 tgaaccaggg acctcaacaa tcacagccac cttcccagca accccttaca tctttaccag
     2641 ctcagcctac tgctcaaagt actagtcaac tgcaggttca agctctaact cagcaacaac
     2701 aatcccctac aaaagctgtg ccggctttgg ggaaaagccc gcctcaccac tctggattcc
     2761 agcagtatca acaggcagat gcctccaaac agctgtggaa tccccctcag gttcaaggcc
     2821 cattagggaa aattatgcct gtgaaacagc cctactacct tcagacccaa gaccccataa
     2881 aactgtttga gccgtcattg caacctcctg taatgcagca gcagcctcta gaaaaaaaaa
     2941 tgaagccttt tcccatggag ccatataacc ataatccctc agaagtcaag gtcccagaat
     3001 tctactggga ttcttcctac agcatggctg ataacagatc tgtaatggca cagcaagcaa
     3061 acatagaccg caggggcaaa cggtcaccag gaatcttccg tccagagcag gatcctgtac
     3121 ccagaatgcc gtttgagaaa tccttattgg agaagccctc agagctcatg tcacattcat
     3181 cctctttcct gtccctcacc ggattctctc tcaatcagga aagataccca aataatagta
     3241 tgttcaatga ggtatatggg aaaaacctga catccagctc caaagcagaa ctcagtccct
     3301 caatggcccc ccaggaaaca tctctgtatt ccctttttga agggactccg tggtctccat
     3361 cacttcctgc cagttcagat cattcaacac cagccagcca gtctcctcat tcctctaacc
     3421 caagcagcct acccagctct cctccaacac acaaccataa ttctgttcca ttctccaatt
     3481 ttggacccat tgggactcca gataacaggg atagaaggac tgcagatcgg tggaaaactg
     3541 ataagccagc catgggtggg tttggcattg attatctctc agcaacgtca tcctctgaga
     3601 gcagttggca tcaggccagc actccgagtg gcacctggac aggccatggc ccttccatgg
     3661 aggattcctc tgctgtcctc atggaaagcc taaagtctat ctggtccagt tccatgatgc
     3721 atcctggacc ttctgctctg gagcagctgt taatgcagca gaagcagaaa cagcaacggg
     3781 gacaaggcac catgaaccct ccacactgag cggccgcttc cctttagtga gggttaatgc
     3841 ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     3901 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     3961 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     4021 agatgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atccgataag
     4081 gatcgatccg ggctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     4141 gcgcagcctg aatggcgaat ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     4201 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     4261 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     4321 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     4381 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     4441 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     4501 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     4561 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttcc
     4621 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgcggatctg
     4681 cgcagcacca tggcctgaaa taacctctga aagaggaact tggttaggta ccttctgagg
     4741 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     4801 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     4861 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     4921 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     4981 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     5041 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagcttga
     5101 ttcttctgac acaacagtct cgaacttaag gctagagcca ccatgattga acaagatgga
     5161 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     5221 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     5281 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     5341 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     5401 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     5461 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     5521 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     5581 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     5641 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     5701 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     5761 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     5821 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     5881 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     5941 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgatggccgc
     6001 aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga atcgatagcg
     6061 ataaggatcc gcgtatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     6121 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     6181 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     6241 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     6301 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     6361 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     6421 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     6481 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     6541 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     6601 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     6661 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     6721 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     6781 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     6841 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     6901 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     6961 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     7021 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     7081 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     7141 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     7201 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     7261 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     7321 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     7381 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     7441 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     7501 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     7561 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     7621 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     7681 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     7741 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     7801 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     7861 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     7921 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     7981 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     8041 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     8101 ttacggttcc tggccttttg ctggcctttt gctcacatgg ctcgacagat ct
//
