LOCUS       sequence-288983-        7028 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsSmg5-del13-266-siRNAres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7028)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 7028)
  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-del13-266-siRNAres_I
FEATURES             Location/Qualifiers
     source          1..7028
                     /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      2968..2977
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2974..2976,2977..2979,2980..3693)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3019..3040)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3280..3299)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3627..3648
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3816..3937
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3853..3872)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3907..3926
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3944..4399)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4081..4102)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4293..4312
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4426..4530
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4532..4889
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4532..4723
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4555..4575)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4740..4875
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4924..5718
                     /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             4924..5718
                     /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(4978..4997)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5588..5607
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5906..5925)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5950..5997
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6326..6914
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6815..6834
                     /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 gcgagacgcg taagaagtgt gagactcgga aactgtctcc tggcaaaaag
      721 cgatgtaaag acattaaaag gttgctagtg aactttatgt atctgcaaag cctcctacag
      781 cccaaaagca gctccgtgga ctcagagctg acctcacttt gccagtcagt cctggaggac
      841 ttcaacctct gcctcttcta cccgccctcc tcacccaacc tcagcctggc cagtgaggat
      901 gaggaggagt atgagagtgg atatgctttc ctcccggacc ttctcatctt tcaaatggtc
      961 atcatccgcc ttatgtgtgt gcacagcttg gagagagcag gatccaagca gtacagtgca
     1021 gccattgcct tcaccctggc cctcttttcc cacctcgtca atcatgtcaa catacggctg
     1081 caggctgagc tggaagaggg cgagaatccc gtcccggcat tccagagtga tggcacagat
     1141 gaaccagagt ccaaggaacc tgtggagaaa gaggaggagc cagatcctga gcctcctcct
     1201 gtaacacccc aagtgggtga gggcagaaag agccgtaagt tctctcgcct ctcctgtctc
     1261 cgccgtcgcc gccacccacc caaagttggt gatgacagtg acctgagtga aggctttgaa
     1321 tcggactcaa gccatgactc agcccgggcc agtgagggct cagacagtgg ctctgacaag
     1381 agtcttgaag gtgggggaac ggcctttgat gctgaaacgg actcggaaat gaatagccag
     1441 gagtcccgat cagacttgga agatatggag gaagaggagg ggacacggtc accaaccctg
     1501 gagccccctc ggggcagatc agaggctccc gattccctca atggcccact gggccccagt
     1561 gaggctagca ttgccagcaa tctacaagcc atgtccaccc agatgttcca gactaagcgc
     1621 tgcttccgac tggcccccac ctttagcaac ctgctcctcc agcccaccac caaccctcat
     1681 acctcggcca gccacaggcc ttgcgtcaat ggggatgtag acaagccttc agagccagcc
     1741 tctgaggagg gctctgagtc ggaggggagt gagtccagtg gacgctcctg tcggaatgag
     1801 cgcagcatcc aggagaagct tcaggtcctg atggccgaag gtctgcttcc tgctgtgaag
     1861 gtatttctag attggctgcg gaccaacccc gacctcatca tcgtgtgtgc gcagagctct
     1921 caaagtctgt ggaaccgcct gtctgtgttg ctgaatctgt tgcctgctgc tggtgaactc
     1981 caggagtctg gcctggcctt gtgtcctgag gtccaagatc ttcttgaagg ttgtgaactg
     2041 cctgacctcc cctctagcct tctgctccca gaggacatgg ctcttcgtaa cctgcccccg
     2101 ctccgagctg cccacagacg ctttaacttt gacacggatc ggcccctgct cagcacctta
     2161 gaggagtcag tggtgcgcat ctgctgcatc cgcagctttg gtcatttcat cgcccgcctg
     2221 caaggcagca tcctgcagtt caacccagag gttggcatct tcgtcagcat tgcccagtct
     2281 gagcaggaga gcctgctgca gcaggcccag gcacagttcc gaatggcaca ggaggaagct
     2341 cgtcggaaca ggctcatgag agacatggct cagctacgac ttcagctcga agtgtctcag
     2401 ctggagggca gcctgcagca gcccaaggcc cagtcagcca tgtctcccta cctcgtccct
     2461 gacacccagg ccctctgcca ccatctccct gtcatccgcc aactggccac cagtggccgc
     2521 ttcattgtca tcatcccaag gacagtgatc gatggcctgg atttgctgaa gaaggaacac
     2581 ccaggggccc gggatgggat tcggtacctg gaggcagagt ttaaaaaagg aaacaggtac
     2641 attcgctgcc agaaagaggt gggaaagagc tttgagcggc ataagctgaa gaggcaggat
     2701 gcagatgcct ggactctcta taagatccta gacagctgca aacagctgac tctggcccag
     2761 ggggcaggtg aggaggatcc gagtggcatg gtgaccatca tcacaggcct tccactggac
     2821 aaccccagcg tgctttcagg ccccatgcag gcagccctgc aggccgctgc ccacgccagt
     2881 gtggacatca agaatgttct ggacttctac aagcagtgga aggaaattgg tacgtcgacg
     2941 gtaccgcggg cccgggatcc accggtcgcc accatggtga gcaagggcga ggagctgttc
     3001 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     3061 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     3121 accaccggca agctgcccgt gccctggccc accctcgtga ccaccttcgg ctacggcctg
     3181 cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     3241 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     3301 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     3361 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     3421 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     3481 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     3541 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc cgccctgagc
     3601 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     3661 atcactctcg gcatggacga gctgtacaag taaagcggcc gcgactctag atcataatca
     3721 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     3781 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     3841 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     3901 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat
     3961 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     4021 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     4081 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     4141 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     4201 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     4261 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     4321 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     4381 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     4441 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     4501 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     4561 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4621 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4681 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     4741 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     4801 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     4861 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     4921 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     4981 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     5041 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     5101 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     5161 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     5221 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     5281 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     5341 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     5401 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     5461 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     5521 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     5581 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     5641 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     5701 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     5761 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     5821 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     5881 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     5941 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     6001 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     6061 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     6121 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     6181 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     6241 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     6301 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     6361 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     6421 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     6481 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     6541 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     6601 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     6661 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     6721 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     6781 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     6841 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     6901 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     6961 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     7021 ccatgcat
//
