LOCUS       sequence-445207-        8488 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsStaufen1-T3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8488)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 8488)
  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-HsStaufen1-T3_V
FEATURES             Location/Qualifiers
     source          1..8488
                     /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    4055..4176
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4066..4085)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4120..4139
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      4362..4817
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4449..4468)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4659..4680
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4934..5291
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4934..5125
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4957..4977)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5142..5277
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5342..6136
                     /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(5396..5415)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6006..6025
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6200..6248
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(6289..6308)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6408..6430
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6468..6486)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6554..6658
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6659..6727,6728..7519)
                     /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(6877..6896)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7690..8278
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8179..8198
                     /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 cgagatgtct
     2281 caagttcaag tgcaagttca gaacccatct gctgctctct cagggagcca aatactgaac
     2341 aagaaccagt ctcttctctc acagcctttg atgagtattc cttctactac tagctctctg
     2401 ccctctgaaa atgcaggtag acccattcaa aactctgctt taccctctgc atctattaca
     2461 tccaccagtg cagctgcaga aagcataacc cctactgtag aactaaatgc actgtgcatg
     2521 aaacttggaa aaaaaccaat gtataagcct gttgaccctt actctcggat gcagtccacc
     2581 tataactaca acatgagagg aggtgcttat cccccgaggt acttttaccc atttccagtt
     2641 ccacctttac tttatcaagt ggaactttct gtgggaggac agcaatttaa tggcaaagga
     2701 aagacaagac aggctgcgaa acacgatgct gctgccaaag cgttgaggat cctgcagaat
     2761 gagcccctgc cagagaggct ggaggtgaat ggaagagaat ccgaagaaga aaatctcaat
     2821 aaatctgaaa taagtcaagt gtttgagatt gcacttaaac ggaacttgcc tgtgaatttc
     2881 gaggtggccc gggagagtgg cccaccccac atgaagaact ttgtgaccaa ggtttcggtt
     2941 ggggagtttg tgggggaagg tgaagggaaa agcaagaaga tttcaaagaa aaatgccgcc
     3001 atagctgttc ttgaggagct gaagaagtta ccgcccctgc ctgcagttga acgagtaaag
     3061 cctagaatca aaaagaaaac aaaacccata gtcaagccac agacaagccc agaatatggc
     3121 caggggatca atccgattag ccgactggcc cagatccagc aggcaaaaaa ggagaaggag
     3181 ccagagtaca cgctcctcac agagcgaggc ctcccgcgcc gcagggagtt tgtgatgcag
     3241 gtgaaggttg gaaaccacac tgcagaagga acgggcacca acaagaaggt ggccaagcgc
     3301 aatgcagccg agaacatgct ggagatcctt ggtttcaaag tcccgcaggc gcagcccacc
     3361 aaacccgcac tcaagtcaga ggagaagaca cccataaaga aaccagggga tggaagaaaa
     3421 gtaacctttt ttgaacctgg ctctggggat gaaaatggga ctagtaataa agaggatgag
     3481 ttcaggatgc cttatctaag tcatcagcag ctgcctgctg gaattcttcc catggtgccc
     3541 gaggtcgccc aggctgtagg agttagtcaa ggacatcaca ccaaagattt taccagggca
     3601 gctccgaatc ctgccaaggc cacggtaact gccatgatag cccgagagtt gttgtatggg
     3661 ggcacctcgc ccacagccga gaccatttta aagaataaca tctcttcagg ccacgtaccc
     3721 catggacctc tcacgagacc ctctgagcaa ctggactatc tttccagagt ccagggattc
     3781 caggttgaat acaaagactt ccccaaaaac aacaagaacg aatttgtatc tcttatcaat
     3841 tgctcctctc agccacctct gatcagccat ggtatcggca aggatgtgga gtcctgccat
     3901 gatatggctg cgctgaacat cttaaagttg ctgtctgagt tggaccaaca aagtacagag
     3961 atgccaagaa caggaaacgg accaatgtct gtgtgtggga ggtgctgagc ggccgcttcc
     4021 ctttagtgag ggttaatgct tcgagcagac atgataagat acattgatga gtttggacaa
     4081 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     4141 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     4201 atgtttcagg ttcaggggga gatgtgggag gttttttaaa gcaagtaaaa cctctacaaa
     4261 tgtggtaaaa tccgataagg atcgatccgg gctggcgtaa tagcgaagag gcccgcaccg
     4321 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gacgcgccct gtagcggcgc
     4381 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     4441 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4501 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     4561 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4621 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4681 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     4741 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     4801 attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     4861 caccgcatac gcggatctgc gcagcaccat ggcctgaaat aacctctgaa agaggaactt
     4921 ggttaggtac cttctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     4981 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     5041 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     5101 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     5161 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     5221 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     5281 gcttttgcaa aaagcttgat tcttctgaca caacagtctc gaacttaagg ctagagccac
     5341 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     5401 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     5461 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     5521 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     5581 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     5641 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     5701 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     5761 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     5821 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     5881 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     5941 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     6001 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     6061 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     6121 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     6181 ctgccatcac gatggccgca ataaaatatc tttattttca ttacatctgt gtgttggttt
     6241 tttgtgtgaa tcgatagcga taaggatccg cgtatggtgc actctcagta caatctgctc
     6301 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     6361 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     6421 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     6481 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     6541 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     6601 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     6661 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     6721 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     6781 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     6841 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     6901 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     6961 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     7021 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     7081 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     7141 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     7201 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     7261 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     7321 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     7381 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     7441 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     7501 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     7561 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     7621 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     7681 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     7741 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     7801 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     7861 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     7921 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     7981 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     8041 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     8101 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     8161 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     8221 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     8281 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatggc
     8341 tcgacagatc ttcaatattg gccattagcc atattattca ttggttatat agcataaatc
     8401 aatattggct attggccatt gcatacgttg tatctatatc ataatatgta catttatatt
     8461 ggctcatgtc caatatgacc gccatgtt
//
