LOCUS       sequence-288985-        7679 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsSmg5-del810-846-siRNAres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7679)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 7679)
  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-del810-846-siRNAres_I
FEATURES             Location/Qualifiers
     source          1..7679
                     /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      3619..3628
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3625..3627,3628..3630,3631..4344)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3670..3691)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3931..3950)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4278..4299
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4467..4588
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4504..4523)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4558..4577
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4595..5050)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4732..4753)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4944..4963
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5077..5181
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5183..5540
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5183..5374
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5206..5226)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5391..5526
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5575..6369
                     /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             5575..6369
                     /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(5629..5648)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6239..6258
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6557..6576)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6601..6648
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6977..7565
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7466..7485
                     /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 gcccagacgc gtcccaaggc ccagtcagcc atgtctccct acctcgtccc tgacacccag
     3121 gccctctgcc accatctccc tgtcatccgc caactggcca ccagtggccg cttcattgtc
     3181 atcatcccaa ggacagtgat cgatggcctg gatttgctga agaaggaaca cccaggggcc
     3241 cgggatggga ttcggtacct ggaggcagag tttaaaaaag gaaacaggta cattcgctgc
     3301 cagaaagagg tgggaaagag ctttgagcgg cataagctga agaggcagga tgcagatgcc
     3361 tggactctct ataagatcct agacagctgc aaacagctga ctctggccca gggggcaggt
     3421 gaggaggatc cgagtggcat ggtgaccatc atcacaggcc ttccactgga caaccccagc
     3481 gtgctttcag gccccatgca ggcagccctg caggccgctg cccacgccag tgtggacatc
     3541 aagaatgttc tggacttcta caagcagtgg aaggaaattg gtacgtcgac ggtaccgcgg
     3601 gcccgggatc caccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg
     3661 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     3721 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     3781 aagctgcccg tgccctggcc caccctcgtg accaccttcg gctacggcct gcagtgcttc
     3841 gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     3901 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     3961 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     4021 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     4081 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
     4141 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     4201 cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccgccctgag caaagacccc
     4261 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     4321 ggcatggacg agctgtacaa gtaaagcggc cgcgactcta gatcataatc agccatacca
     4381 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     4441 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     4501 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     4561 gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta
     4621 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     4681 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     4741 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     4801 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     4861 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     4921 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     4981 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     5041 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     5101 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     5161 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     5221 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     5281 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     5341 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     5401 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     5461 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     5521 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     5581 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     5641 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     5701 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     5761 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     5821 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     5881 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     5941 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     6001 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     6061 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     6121 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     6181 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     6241 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     6301 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     6361 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     6421 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     6481 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     6541 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     6601 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     6661 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     6721 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     6781 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     6841 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6901 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6961 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     7021 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     7081 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     7141 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     7201 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     7261 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     7321 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     7381 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     7441 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     7501 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     7561 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     7621 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
