LOCUS       sequence-288981-        8455 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsSmg7-del499-632-siRNAres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8455)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 8455)
  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-del499-632-siRNAres_I
FEATURES             Location/Qualifiers
     source          1..8455
                     /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    4159..4280
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4170..4189)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4224..4243
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      4466..4921
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4553..4572)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4763..4784
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5038..5395
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5038..5229
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5061..5081)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5246..5381
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5446..6240
                     /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(5500..5519)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6110..6129
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6304..6352
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(6393..6412)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6512..6534
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6572..6590)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6658..6762
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6763..6831,6832..7623)
                     /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(6981..7000)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7794..8382
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8283..8302
                     /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 ctctcaagta
     2461 ttagtgcgac accacttcca 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 cccagtacgc gtatgggctc aggttacacc ttcccagctg
     2761 gtgtttctgt cccaggaacc tttcttcggc ctacagctca ctctccagca ggaaaccagg
     2821 tgcaagctgg gaaacagtcc cacattcctt acagccagca acggccctct ggaccagggc
     2881 caatgaacca gggacctcaa caatcacagc caccttccca gcaacccctt acatctttac
     2941 cagctcagcc tactgctcaa agtactagtc aactgcaggt tcaagctcta actcagcaac
     3001 aacaatcccc tacaaaagct gtgccggctt tggggaaaag cccgcctcac cactctggat
     3061 tccagcagta tcaacaggca gatgcctcca aacagctgtg gaatccccct caggttcaag
     3121 gcccattagg gaaaattatg cctgtgaaac agccctacta ccttcagacc caagacccca
     3181 taaaactgtt tgagccgtca ttgcaacctc ctgtaatgca gcagcagcct ctagaaaaaa
     3241 aaatgaagcc ttttcccatg gagccatata accataatcc ctcagaagtc aaggtcccag
     3301 aattctactg ggattcttcc tacagcatgg ctgataacag atctgtaatg gcacagcaag
     3361 caaacataga ccgcaggggc aaacggtcac caggaatctt ccgtccagag caggatcctg
     3421 tacccagaat gccgtttgag aaatccttat tggagaagcc ctcagagctc atgtcacatt
     3481 catcctcttt cctgtccctc accggattct ctctcaatca ggaaagatac ccaaataata
     3541 gtatgttcaa tgaggtatat gggaaaaacc tgacatccag ctccaaagca gaactcagtc
     3601 cctcaatggc cccccaggaa acatctctgt attccctttt tgaagggact ccgtggtctc
     3661 catcacttcc tgccagttca gatcattcaa caccagccag ccagtctcct cattcctcta
     3721 acccaagcag cctacccagc tctcctccaa cacacaacca taattctgtt ccattctcca
     3781 attttggacc cattgggact ccagataaca gggatagaag gactgcagat cggtggaaaa
     3841 ctgataagcc agccatgggt gggtttggca ttgattatct ctcagcaacg tcatcctctg
     3901 agagcagttg gcatcaggcc agcactccga gtggcacctg gacaggccat ggcccttcca
     3961 tggaggattc ctctgctgtc ctcatggaaa gcctaaagtc tatctggtcc agttccatga
     4021 tgcatcctgg accttctgct ctggagcagc tgttaatgca gcagaagcag aaacagcaac
     4081 ggggacaagg caccatgaac cctccacact gagcggccgc ttccctttag tgagggttaa
     4141 tgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     4201 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     4261 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     4321 gggagatgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatccgat
     4381 aaggatcgat ccgggctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     4441 gttgcgcagc ctgaatggcg aatggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     4501 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     4561 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     4621 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     4681 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     4741 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     4801 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     4861 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     4921 tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgcggat
     4981 ctgcgcagca ccatggcctg aaataacctc tgaaagagga acttggttag gtaccttctg
     5041 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     5101 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     5161 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     5221 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     5281 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     5341 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     5401 tgattcttct gacacaacag tctcgaactt aaggctagag ccaccatgat tgaacaagat
     5461 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     5521 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     5581 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     5641 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     5701 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     5761 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     5821 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     5881 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     5941 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     6001 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     6061 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     6121 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     6181 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     6241 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgatggc
     6301 cgcaataaaa tatctttatt ttcattacat ctgtgtgttg gttttttgtg tgaatcgata
     6361 gcgataagga tccgcgtatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     6421 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     6481 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     6541 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     6601 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     6661 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     6721 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     6781 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     6841 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     6901 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     6961 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     7021 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     7081 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     7141 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     7201 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     7261 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     7321 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     7381 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     7441 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     7501 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     7561 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     7621 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     7681 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     7741 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     7801 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     7861 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     7921 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     7981 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     8041 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     8101 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     8161 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     8221 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     8281 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     8341 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     8401 tttttacggt tcctggcctt ttgctggcct tttgctcaca tggctcgaca gatct
//
