LOCUS       sequence-445208-        8245 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 8245)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 8245)
  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-T2_V
FEATURES             Location/Qualifiers
     source          1..8245
                     /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    3812..3933
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3823..3842)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3877..3896
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      4119..4574
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4206..4225)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4416..4437
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4691..5048
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4691..4882
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4714..4734)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4899..5034
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5099..5893
                     /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(5153..5172)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5763..5782
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5957..6005
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(6046..6065)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6165..6187
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6225..6243)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6311..6415
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6416..6484,6485..7276)
                     /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(6634..6653)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7447..8035
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7936..7955
                     /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 cgagatgaaa
     2281 cttggaaaaa aaccaatgta taagcctgtt gacccttact ctcggatgca gtccacctat
     2341 aactacaaca tgagaggagg tgcttatccc ccgaggtact tttacccatt tccagttcca
     2401 cctttacttt atcaagtgga actttctgtg ggaggacagc aatttaatgg caaaggaaag
     2461 acaagacagg ctgcgaaaca cgatgctgct gccaaagcgt tgaggatcct gcagaatgag
     2521 cccctgccag agaggctgga ggtgaatgga agagaatccg aagaagaaaa tctcaataaa
     2581 tctgaaataa gtcaagtgtt tgagattgca cttaaacgga acttgcctgt gaatttcgag
     2641 gtggcccggg agagtggccc accccacatg aagaactttg tgaccaaggt ttcggttggg
     2701 gagtttgtgg gggaaggtga agggaaaagc aagaagattt caaagaaaaa tgccgccata
     2761 gctgttcttg aggagctgaa gaagttaccg cccctgcctg cagttgaacg agtaaagcct
     2821 agaatcaaaa agaaaacaaa acccatagtc aagccacaga caagcccaga atatggccag
     2881 gggatcaatc cgattagccg actggcccag atccagcagg caaaaaagga gaaggagcca
     2941 gagtacacgc tcctcacaga gcgaggcctc ccgcgccgca gggagtttgt gatgcaggtg
     3001 aaggttggaa accacactgc agaaggaacg ggcaccaaca agaaggtggc caagcgcaat
     3061 gcagccgaga acatgctgga gatccttggt ttcaaagtcc cgcaggcgca gcccaccaaa
     3121 cccgcactca agtcagagga gaagacaccc ataaagaaac caggggatgg aagaaaagta
     3181 accttttttg aacctggctc tggggatgaa aatgggacta gtaataaaga ggatgagttc
     3241 aggatgcctt atctaagtca tcagcagctg cctgctggaa ttcttcccat ggtgcccgag
     3301 gtcgcccagg ctgtaggagt tagtcaagga catcacacca aagattttac cagggcagct
     3361 ccgaatcctg ccaaggccac ggtaactgcc atgatagccc gagagttgtt gtatgggggc
     3421 acctcgccca cagccgagac cattttaaag aataacatct cttcaggcca cgtaccccat
     3481 ggacctctca cgagaccctc tgagcaactg gactatcttt ccagagtcca gggattccag
     3541 gttgaataca aagacttccc caaaaacaac aagaacgaat ttgtatctct tatcaattgc
     3601 tcctctcagc cacctctgat cagccatggt atcggcaagg atgtggagtc ctgccatgat
     3661 atggctgcgc tgaacatctt aaagttgctg tctgagttgg accaacaaag tacagagatg
     3721 ccaagaacag gaaacggacc aatgtctgtg tgtgggaggt gctgagcggc cgcttccctt
     3781 tagtgagggt taatgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc
     3841 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     3901 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     3961 tttcaggttc agggggagat gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt
     4021 ggtaaaatcc gataaggatc gatccgggct ggcgtaatag cgaagaggcc cgcaccgatc
     4081 gcccttccca acagttgcgc agcctgaatg gcgaatggac gcgccctgta gcggcgcatt
     4141 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     4201 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     4261 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     4321 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     4381 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     4441 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     4501 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     4561 aacgcttaca atttcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     4621 cgcatacgcg gatctgcgca gcaccatggc ctgaaataac ctctgaaaga ggaacttggt
     4681 taggtacctt ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     4741 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4801 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4861 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4921 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4981 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     5041 tttgcaaaaa gcttgattct tctgacacaa cagtctcgaa cttaaggcta gagccaccat
     5101 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     5161 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     5221 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     5281 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     5341 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     5401 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     5461 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     5521 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     5581 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     5641 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     5701 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     5761 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     5821 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     5881 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     5941 ccatcacgat ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt
     6001 gtgtgaatcg atagcgataa ggatccgcgt atggtgcact ctcagtacaa tctgctctga
     6061 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     6121 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     6181 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     6241 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     6301 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     6361 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     6421 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     6481 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     6541 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     6601 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     6661 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     6721 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     6781 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     6841 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     6901 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     6961 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     7021 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     7081 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     7141 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     7201 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     7261 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     7321 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     7381 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     7441 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     7501 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     7561 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     7621 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     7681 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     7741 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     7801 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     7861 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     7921 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     7981 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     8041 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatggctcg
     8101 acagatcttc aatattggcc attagccata ttattcattg gttatatagc ataaatcaat
     8161 attggctatt ggccattgca tacgttgtat ctatatcata atatgtacat ttatattggc
     8221 tcatgtccaa tatgaccgcc atgtt
//
