LOCUS       sequence-445205-        8467 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsStaufen2-T1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8467)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 8467)
  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-V5-SBP-MBP-HsStaufen2-T1_V
FEATURES             Location/Qualifiers
     source          1..8467
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        77..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..592
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     543..563
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     589..613
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          753..885
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     868..887
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        930..948
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      961..970
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             970..1011
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1033..1146
                     /codon_start=1
                     /product="streptavidin-binding peptide"
                     /label=SBP
                     /note="selected from a peptide library; binds streptavidin 
                     with nanomolar affinity (Keefe et al., 2001)"
                     /translation="MDEKTTGWGGGHVVEGLAGELEQLRARLEHHPQGQREP"
     regulatory      2128..2137
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4034..4155
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4045..4064)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4099..4118
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      4341..4796
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4428..4447)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4638..4659
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4913..5270
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4913..5104
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4936..4956)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5121..5256
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5321..6115
                     /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(5375..5394)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5985..6004
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6179..6227
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(6268..6287)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6387..6409
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6447..6465)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6533..6637
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6638..6706,6707..7498)
                     /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(6856..6875)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7669..8257
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8158..8177
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggcattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctgcgatcgc ccgccccgtt
      541 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt
      601 gaaccgtcag atcactagaa gctttattgc ggtagtttat cacagttaaa ttgctaacgc
      661 agtcagtgct tctgacacaa cagtctcgaa cttaagctgc agtgactctc ttaaggtagc
      721 cttgcagaag ttggtcgtga ggcactgggc aggtaagtat caaggttaca agacaggttt
      781 aaggagacca atagaaactg ggcttgtcga gacagagaag actcttgcgt ttctgatagg
      841 cacctattgg tcttactgac atccactttg cctttctctc cacaggtgtc cactcccagt
      901 tcaattacag ctcttaaggc tagagtactt aatacgactc actataggct agcaattgcg
      961 gccaccatgg gtaagcctat ccctaaccct ctcctcggtc tcgattctac gcccggaagc
     1021 ggtagcggta ccatggacga gaagaccacc ggctgggggg gcggccacgt ggtggagggc
     1081 ctggccggcg agctggagca gctgcgggcc aggctggagc accaccctca gggccagcgg
     1141 gagccctccg gcggctgcaa gcccctcgac atgaaaatcg aagaaggtaa actggtaatc
     1201 tggattaacg gcgataaagg ctataacggt ctcgctgaag tcggtaagaa attcgagaaa
     1261 gataccggaa ttaaagtcac cgttgagcat ccggataaac tggaagagaa attcccgcag
     1321 gttgcggcaa ctggcgatgg ccctgacatt atcttctggg cacacgaccg ctttggtggc
     1381 tacgctcaat ctggcctgtt ggctgaaatc accccggaca aagcgttcca ggacaagctg
     1441 tatccgttta cctgggatgc cgtacgttac aacggcaagc tgattgctta cccgatcgct
     1501 gttgaagcgt tatcgctgat ttataacaaa gatctgctgc cgaacccgcc aaaaacctgg
     1561 gaagagatcc cggcgctgga taaagaactg aaagcgaaag gtaagagcgc gctgatgttc
     1621 aacctgcaag aaccgtactt cacctggccg ctgattgctg ctgacggggg ttatgcgttc
     1681 aagtatgaaa acggcaagta cgacattaaa gacgtgggcg tggataacgc tggcgcgaaa
     1741 gcgggtctga ccttcctggt tgacctgatt aaaagcaaac acatgaatgc agacaccgat
     1801 tactccatcg cagaagctgc ctttaataaa ggcgaaacag cgatgaccat caacggcccg
     1861 tgggcatggt ccaacatcga caccagcaaa gtgaattatg gtgtaacggt actgccgacc
     1921 ttcaagggtc aaccatccaa accgttcgtt ggcgtgctga gcgcaggtat taacgccgcc
     1981 agtccgaaca aagagctggc gaaagagttc ctcgaaaact atctgctgac tgatgaaggt
     2041 ctggaagcgg ttaataaaga caaaccgctg ggtgccgtag cgctgaagtc ttacgaggaa
     2101 gagttggcga aagatccacg tattgccgcc accatggaaa acgcccagaa aggtgaaatc
     2161 atgccgaaca tcccgcagat gtccgctttc tggtatgccg tgcgtactgc ggtgatcaac
     2221 gccgccagcg gtcgtcagac tgtcgatgaa gccctgaaag accctccact cgagatggca
     2281 aacccaaaag agaaaactgc aatgtgtctg gtaaatgagt tagcccgttt caatagagtc
     2341 caaccccagt ataaacttct gaatgaaaga gggcctgctc attcaaagat gttctcagtg
     2401 cagctgagtc ttggtgagca gacatgggaa tccgaaggca gcagtataaa gaaggctcag
     2461 caggctgttg ccaataaagc tttgactgaa tctacgcttc ccaaaccagt tcagaagcca
     2521 cccaaaagta atgttaacaa taacccaggc agtataactc caactgtgga actgaatggg
     2581 cttgctatga aaaggggaga gcctgccatc tacaggccat tagatccaaa gccattccca
     2641 aattatagag ctaattacaa ctttcggggc atgtacaatc agaggtatca ttgcccagtg
     2701 cctaagatct tttatgttca gctcactgta ggaaataatg aattttttgg ggaaggaaag
     2761 actcgacaag ctgctagaca caatgctgca atgaaagccc tccaagcact gcagaatgaa
     2821 cctattccag aaagatctcc tcagaatggt gaatcaggaa aggatgtgga tgatgacaaa
     2881 gatgcaaata agtctgagat cagcttagtg tttgaaattg ctctgaagcg aaatatgcct
     2941 gtcagttttg aggttattaa agaaagtgga ccaccacata tgaaaagctt tgttactcga
     3001 gtgtcagtag gagagttctc tgcagaagga gaaggaaata gcaaaaaact ctccaagaag
     3061 cgcgctgcga ccaccgtctt acaggagctt aaaaaacttc cacctcttcc tgtggtggaa
     3121 aagccaaaac tattttttaa aaaacgccct aaaacaatag taaaggccgg accagaatat
     3181 ggccaaggga tgaaccctat tagccgcctg gcgcaaattc aacaggccaa aaaggaaaag
     3241 gagccggatt atgttttgct ttcagaaaga ggaatgcctc gacgtcgaga atttgtgatg
     3301 caggtgaagg taggcaatga agttgctaca ggaacaggac ctaataaaaa gatagccaaa
     3361 aaaaatgctg cagaagcaat gctgttacaa cttggttata aagcatccac taatcttcag
     3421 gatcaacttg agaagacagg ggaaaacaaa ggatggagtg gtccaaagcc tgggtttcct
     3481 gaaccaacaa ataatactcc aaaaggaatt cttcatttgt ctcctgatgt ttatcaagag
     3541 atggaagcca gccgccacaa agtaatctct ggcactactc taggctattt gtcacccaaa
     3601 gatatgaacc aaccttcaag ctctttcttc agtatatctc ccacatcgaa tagttcagct
     3661 acaattgcca gggaactcct tatgaatgga acatcttcta cagctgaagc cataggttta
     3721 aaaggaagtt ctcctactcc cccttgttct ccagtacaac cttcaaaaca actggaatat
     3781 ttagcaagga ttcaaggctt tcaggcagcc ttaagtgcct tgaaacaatt ttctgaacaa
     3841 ggactggatc caatcgatgg agcaatgaat atcgaaaaag gttctcttga aaaacaagcc
     3901 aagcatctga gagagaaagc ggacaataac caggcacccc cgggctccat cgctcaggac
     3961 tgcaagaaat caaactcggc cgtctaggcg gccgcttccc tttagtgagg gttaatgctt
     4021 cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     4081 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     4141 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     4201 atgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtaaaat ccgataagga
     4261 tcgatccggg ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     4321 gcagcctgaa tggcgaatgg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     4381 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     4441 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     4501 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     4561 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     4621 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     4681 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     4741 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttcctg
     4801 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg cggatctgcg
     4861 cagcaccatg gcctgaaata acctctgaaa gaggaacttg gttaggtacc ttctgaggcg
     4921 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     4981 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     5041 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     5101 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     5161 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     5221 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagcttgatt
     5281 cttctgacac aacagtctcg aacttaaggc tagagccacc atgattgaac aagatggatt
     5341 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     5401 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     5461 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     5521 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     5581 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     5641 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     5701 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     5761 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     5821 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     5881 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     5941 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     6001 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     6061 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     6121 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg atggccgcaa
     6181 taaaatatct ttattttcat tacatctgtg tgttggtttt ttgtgtgaat cgatagcgat
     6241 aaggatccgc gtatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     6301 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     6361 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     6421 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     6481 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     6541 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     6601 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     6661 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     6721 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     6781 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     6841 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     6901 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     6961 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     7021 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     7081 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     7141 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     7201 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     7261 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     7321 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     7381 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     7441 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     7501 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     7561 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     7621 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     7681 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     7741 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     7801 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     7861 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     7921 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     7981 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     8041 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     8101 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     8161 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     8221 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     8281 acggttcctg gccttttgct ggccttttgc tcacatggct cgacagatct tcaatattgg
     8341 ccattagcca tattattcat tggttatata gcataaatca atattggcta ttggccattg
     8401 catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc aatatgaccg
     8461 ccatgtt
//
