LOCUS       sequence-420958-        7361 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsStaufen1-T2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7361)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 7361)
  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: pcDNA3.1-MS2-HA-HsStaufen1-T2_V
FEATURES             Location/Qualifiers
     source          1..7361
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(36..55)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        227..606
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        607..810
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     761..781
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     855..874
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        855..873
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      887..896
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1325..1351
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(2906..2924)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2907..2924)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2944..2961)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2950..3174
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3220..3648
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3307..3326)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3517..3538
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3657..3677)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3662..3991
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3842..3977
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3904..3923
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4058..4852
                     /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(4112..4131)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4722..4741
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5026..5147
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5063..5082)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5117..5136
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5196..5212)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5196..5212)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5209..5231)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5220..5236
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5244..5250,5251..5268,5269..5274))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5289..5310
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5427..5444)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5598..6186)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5678..5697)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6357..7148,7149..7217))
                     /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     6980..6999
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7218..7322)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag
       61 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
      121 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt
      181 aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca ttgattattg
      241 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
      301 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      361 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      421 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      481 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      541 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      601 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
      661 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
      721 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
      781 gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc cactgcttac
      841 tggcttatcg aaattaatac gactcactat agggagaccc aagcttgcca ccatggcttc
      901 taactttact cagttcgttc tcgtcgacaa tggcggaact ggcgacgtga ctgtcgcccc
      961 aagcaacttc gctaacgggg tcgctgaatg gatcagctct aactcgcgat cacaggctta
     1021 caaagtaacc tgtagcgttc gtcagagctc tgcgcagaat cgcaaataca ccatcaaagt
     1081 cgaggtgcct aaagtggcaa cccagactgt tggtggtgaa gagcttcctg tagccggatg
     1141 gagatcttac ttaaatatgg aactaaccat tccaattttc gccacgaatt ccgactgcga
     1201 gcttattgtt aaggcaatgc aaggtctcct aaaagatgga aacccgattc cctcggccat
     1261 cgcggccaac tccggcatct acggaggtgg aggtggatct ggtggatctc ctccacatat
     1321 ggcttaccca tacgatgttc cagattacgc tcctccaaag cttggatccg aattcccggg
     1381 tcgactcgag atgaaacttg gaaaaaaacc aatgtataag cctgttgacc cttactctcg
     1441 gatgcagtcc acctataact acaacatgag aggaggtgct tatcccccga ggtactttta
     1501 cccatttcca gttccacctt tactttatca agtggaactt tctgtgggag gacagcaatt
     1561 taatggcaaa ggaaagacaa gacaggctgc gaaacacgat gctgctgcca aagcgttgag
     1621 gatcctgcag aatgagcccc tgccagagag gctggaggtg aatggaagag aatccgaaga
     1681 agaaaatctc aataaatctg aaataagtca agtgtttgag attgcactta aacggaactt
     1741 gcctgtgaat ttcgagaggt ggcccgggag agtggcccac cccacatgaa gaactttgtg
     1801 accaaggttt cggttgggga gtttgtgggg gaaggtgaag ggaaaagcaa gaagatttca
     1861 aagaaaaatg ccgccatagc tgttcttgag gagctgaaga agttaccgcc cctgcctgca
     1921 gttgaacgag taaagcctag aatcaaaaag aaaacaaaac ccatagtcaa gccacagaca
     1981 agcccagaat atggccaggg gatcaatccg attagccgac tggcccagat ccagcaggca
     2041 aaaaaggaga aggagccaga gtacacgctc ctcacagagc gaggcctccc gcgccgcagg
     2101 gagtttgtga tgcaggtgaa ggttggaaac cacactgcag aaggaacggg caccaacaag
     2161 aaggtggcca agcgcaatgc agccgagaac atgctggaga tccttggttt caaagtcccg
     2221 caggcgcagc ccaccaaacc cgcactcaag tcagaggaga agacacccat aaagaaacca
     2281 ggggatggaa gaaaagtaac cttttttgaa cctggctctg gggatgaaaa tgggactagt
     2341 aataaagagg atgagttcag gatgccttat ctaagtcatc agcagctgcc tgctggaatt
     2401 cttcccatgg tgcccgaggt cgcccaggct gtaggagtta gtcaaggaca tcacaccaaa
     2461 gattttacca gggcagctcc gaatcctgcc aaggccacgg taactgccat gatagcccga
     2521 gagttgttgt atgggggcac ctcgcccaca gccgagacca ttttaaagaa taacatctct
     2581 tcaggccacg taccccatgg acctctcacg agaccctctg agcaactgga ctatctttcc
     2641 agagtccagg gattccaggt tgaatacaaa gacttcccca aaaacaacaa gaacgaattt
     2701 gtatctctta tcaattgctc ctctcagcca cctctgatca gccatggtat cggcaaggat
     2761 gtggagtcct gccatgatat ggctgcgctg aacatcttaa agttgctgtc tgagttggac
     2821 caacaaagta cagagatgcc aagaacagga aacggaccaa tgtctgtgtg tgggaggtgc
     2881 tgagcggccg cctagagggc cctattctat agtgtcacct aaatgctaga gctcgctgat
     2941 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     3001 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     3061 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     3121 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     3181 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     3241 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     3301 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3361 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     3421 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     3481 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     3541 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     3601 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     3661 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3721 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     3781 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     3841 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     3901 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     3961 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     4021 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     4081 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     4141 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     4201 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     4261 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     4321 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     4381 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     4441 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     4501 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     4561 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     4621 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     4681 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     4741 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     4801 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     4861 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     4921 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     4981 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     5041 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     5101 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     5161 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     5221 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     5281 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     5341 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     5401 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     5461 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     5521 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5581 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5641 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5701 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5761 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5821 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5881 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5941 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6001 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     6061 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6121 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6181 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6241 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6301 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     6361 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     6421 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     6481 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     6541 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     6601 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     6661 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     6721 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     6781 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     6841 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     6901 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     6961 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     7021 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     7081 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     7141 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     7201 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     7261 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     7321 cgcacatttc cccgaaaagt gccacctgac gtcgacggat c
//
