LOCUS       sequence-288987-        7784 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsSmg5-siRNAres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7784)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 7784)
  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: pEYFP-N1-HsSmg5-siRNAres_I
FEATURES             Location/Qualifiers
     source          1..7784
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3724..3733
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3730..3732,3733..3735,3736..4449)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3775..3796)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4036..4055)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4383..4404
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4572..4693
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4609..4628)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4663..4682
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4700..5155)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4837..4858)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5049..5068
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5182..5286
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5288..5645
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5288..5479
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5311..5331)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5496..5631
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5680..6474
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5680..6474
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5734..5753)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6344..6363
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6662..6681)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6706..6753
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7082..7670
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7571..7590
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attcgcaaca tgagccaagg cccccccaca
      661 ggggagagca gcgagcccga agcaaaagtc ctccacacta agcggcttta ccgggctgtg
      721 gtggaggctg tgcatcgact tgacctcatc ctttgcaaca aaactgctta tcaagaagta
      781 ttcaaaccag aaaacattag cctgaggaac aagctgcgtg agctctgcgt caagcttatg
      841 ttcctgcacc cagtggacta tgggagaaag gctgaggagc tgctgtggag aagggtatac
      901 tatgaagtta tccagcttat caagactaac aaaaagcaca tccacagccg gagcactttg
      961 gaatgtgcct acaggacgca cctggttgct ggtattggct tctaccagca tctccttctc
     1021 tatatccagt cccactacca gctggaactg cagtgctgca tcgactggac ccatgtcact
     1081 gaccccctca taggatgcaa gaagccagtg tctgcctcag ggaaggagat ggattgggca
     1141 cagatggcat gtcaccgatg tctggtgtat ctgggggatt tgtcccgata tcagaatgaa
     1201 ttagctggcg tagataccga gctgctagcc gagagatttt actaccaagc cctgtcagta
     1261 gctcctcaga ttggaatgcc cttcaatcag ctgggcaccc tggcaggcag caagtactat
     1321 aatgtggaag ccatgtattg ctacctgcgc tgcatccagt cagaagtgtc ctttgaggga
     1381 gcctatggga acctcaagcg gctgtatgac aaggcagcca aaatgtacca ccaactgaag
     1441 aagtgtgaga ctcggaaact gtctcctggc aaaaagcgat gtaaagacat taaaaggttg
     1501 ctagtgaact ttatgtatct gcaaagcctc ctacagccca aaagcagctc cgtggactca
     1561 gagctgacct cactttgcca gtcagtcctg gaggacttca acctctgcct cttctacccg
     1621 ccctcctcac ccaacctcag cctggccagt gaggatgagg aggagtatga gagtggatat
     1681 gctttcctcc cggaccttct catctttcaa atggtcatca tccgccttat gtgtgtgcac
     1741 agcttggaga gagcaggatc caagcagtac agtgcagcca ttgccttcac cctggccctc
     1801 ttttcccacc tcgtcaatca tgtcaacata cggctgcagg ctgagctgga agagggcgag
     1861 aatcccgtcc cggcattcca gagtgatggc acagatgaac cagagtccaa ggaacctgtg
     1921 gagaaagagg aggagccaga tcctgagcct cctcctgtaa caccccaagt gggtgagggc
     1981 agaaagagcc gtaagttctc tcgcctctcc tgtctccgcc gtcgccgcca cccacccaaa
     2041 gttggtgatg acagtgacct gagtgaaggc tttgaatcgg actcaagcca tgactcagcc
     2101 cgggccagtg agggctcaga cagtggctct gacaagagtc ttgaaggtgg gggaacggcc
     2161 tttgatgctg aaacggactc ggaaatgaat agccaggagt cccgatcaga cttggaagat
     2221 atggaggaag aggaggggac acggtcacca accctggagc cccctcgggg cagatcagag
     2281 gctcccgatt ccctcaatgg cccactgggc cccagtgagg ctagcattgc cagcaatcta
     2341 caagccatgt ccacccagat gttccagact aagcgctgct tccgactggc ccccaccttt
     2401 agcaacctgc tcctccagcc caccaccaac cctcatacct cggccagcca caggccttgc
     2461 gtcaatgggg atgtagacaa gccttcagag ccagcctctg aggagggctc tgagtcggag
     2521 gggagtgagt ccagtggacg ctcctgtcgg aatgagcgca gcatccagga gaagcttcag
     2581 gtcctgatgg ccgaaggtct gcttcctgct gtgaaggtat ttctagattg gctgcggacc
     2641 aaccccgacc tcatcatcgt gtgtgcgcag agctctcaaa gtctgtggaa ccgcctgtct
     2701 gtgttgctga atctgttgcc tgctgctggt gaactccagg agtctggcct ggccttgtgt
     2761 cctgaggtcc aagatcttct tgaaggttgt gaactgcctg acctcccctc tagccttctg
     2821 ctcccagagg acatggctct tcgtaacctg cccccgctcc gagctgccca cagacgcttt
     2881 aactttgaca cggatcggcc cctgctcagc accttagagg agtcagtggt gcgcatctgc
     2941 tgcatccgca gctttggtca tttcatcgcc cgcctgcaag gcagcatcct gcagttcaac
     3001 ccagaggttg gcatcttcgt cagcattgcc cagtctgagc aggagagcct gctgcagcag
     3061 gcccaggcac agttccgaat ggcacaggag gaagctcgtc ggaacaggct catgagagac
     3121 atggctcagc tacgacttca gctcgaagtg tctcagctgg agggcagcct gcagcagccc
     3181 aaggcccagt cagccatgtc tccctacctc gtccctgaca cccaggccct ctgccaccat
     3241 ctccctgtca tccgccaact ggccaccagt ggccgcttca ttgtcatcat cccaaggaca
     3301 gtgatcgatg gcctggattt gctgaagaag gaacacccag gggcccggga tgggattcgg
     3361 tacctggagg cagagtttaa aaaaggaaac aggtacattc gctgccagaa agaggtggga
     3421 aagagctttg agcggcataa gctgaagagg caggatgcag atgcctggac tctctataag
     3481 atcctagaca gctgcaaaca gctgactctg gcccaggggg caggtgagga ggatccgagt
     3541 ggcatggtga ccatcatcac aggccttcca ctggacaacc ccagcgtgct ttcaggcccc
     3601 atgcaggcag ccctgcaggc cgctgcccac gccagtgtgg acatcaagaa tgttctggac
     3661 ttctacaagc agtggaagga aattggtacg tcgacggtac cgcgggcccg ggatccaccg
     3721 gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     3781 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     3841 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     3901 tggcccaccc tcgtgaccac cttcggctac ggcctgcagt gcttcgcccg ctaccccgac
     3961 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     4021 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     4081 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     4141 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     4201 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     4261 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     4321 gacaaccact acctgagcta ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     4381 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     4441 tacaagtaaa gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt
     4501 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     4561 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     4621 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     4681 tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     4741 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     4801 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     4861 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     4921 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     4981 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     5041 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     5101 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt
     5161 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     5221 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     5281 aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     5341 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     5401 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     5461 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     5521 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     5581 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     5641 gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     5701 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     5761 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     5821 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta
     5881 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     5941 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     6001 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     6061 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     6121 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     6181 gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc
     6241 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     6301 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     6361 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     6421 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     6481 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     6541 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     6601 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa
     6661 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     6721 agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct
     6781 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
     6841 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
     6901 cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc
     6961 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     7021 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     7081 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     7141 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     7201 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     7261 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     7321 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     7381 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     7441 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     7501 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     7561 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     7621 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     7681 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     7741 cgttatcccc tgattctgtg gataaccgta ttaccgccat gcat
//
