LOCUS       sequence-420985-        7602 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Mammalian Expression of HsStaufen1-T3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7602)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 7602)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-T3_V
FEATURES             Location/Qualifiers
     source          1..7602
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      707..1295
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1196..1215
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1449..1466
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1583..1604
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1619..1624,1625..1642,1643..1649)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1657..1673
                     /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)."
     primer_bind     1662..1684
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1681..1697
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1681..1697
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1746..1867
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1757..1776)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1811..1830
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2041..2835)
                     /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(2152..2171)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2762..2781
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2902..3231)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2916..3051
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2970..2989)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     3216..3236
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(3245..3673)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3355..3376)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3567..3586
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    3719..3943
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3932..3949
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        3969..3987
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3969..3986
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             complement(5783..5809)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      6238..6247
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(6260..6279)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6261..6279)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(6324..6527)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6353..6373)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6528..6907
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     7079..7098
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        7173..7277
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7278..7346,7347..7602,1..536)
                     /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(7496..7515)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
       61 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
      121 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
      181 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
      241 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
      301 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
      361 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
      421 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
      481 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
      541 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
      601 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
      661 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
      721 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
      781 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
      841 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
      901 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
      961 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     1021 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     1081 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     1141 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     1201 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     1261 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     1321 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     1381 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     1441 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     1501 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     1561 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
     1621 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     1681 caggaaacag ctatgaccat gattacgcca agctctagct agaggtcgac ggtatacaga
     1741 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     1801 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     1861 acaagttggg gtgggcgaag aactccagca tgagatcccc gcgctggagg atcatccagc
     1921 cggcgtcccg gaaaacgatt ccgaagccca acctttcata gaaggcggcg gtggaatcga
     1981 aatctcgtga tggcaggttg ggcgtcgctt ggtcggtcat ttcgaacccc agagtcccgc
     2041 tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat
     2101 accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg
     2161 ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac agtcgatgaa
     2221 tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc catgggtcac
     2281 gacgagatcc tcgccgtcgg gcatgcgcgc cttgagcctg gcgaacagtt cggctggcgc
     2341 gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt
     2401 acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag
     2461 cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg
     2521 agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc
     2581 agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg
     2641 cgctgcctcg tcctgcagtt cattcagggc accggacagg tcggtcttga caaaaagaac
     2701 cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg
     2761 tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc
     2821 atcttgttca atcatgcgaa acgatcctca tcctgtctct tgatcagatc cgaaaatgga
     2881 tatacaagct cccgggagct ttttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta
     2941 cttctggaat agctcagagg cagaggcggc ctcggcctct gcataaataa aaaaaattag
     3001 tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt
     3061 taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
     3121 tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat
     3181 acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca cattccacag
     3241 aattaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     3301 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     3361 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     3421 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     3481 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     3541 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     3601 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     3661 ctacagggcg cgtggggata ccccctagag ccccagctgg ttctttccgc ctcagaagcc
     3721 atagagccca ccgcatcccc agcatgcctg ctattgtctt cccaatcctc ccccttgctg
     3781 tcctgcccca ccccaccccc cagaatagaa tgacacctac tcagacaatg cgatgcaatt
     3841 tcctcatttt attaggaaag gacagtggga gtggcacctt ccagggtcaa ggaaggcacg
     3901 ggggaggggc aaacaacaga tggctggcaa ctagaaggca cagtcgaggc tgatcagcga
     3961 gctctagcat ttaggtgaca ctatagaata gggccctcta ggcggccgct cagcacctcc
     4021 cacacacaga cattggtccg tttcctgttc ttggcatctc tgtactttgt tggtccaact
     4081 cagacagcaa ctttaagatg ttcagcgcag ccatatcatg gcaggactcc acatccttgc
     4141 cgataccatg gctgatcaga ggtggctgag aggagcaatt gataagagat acaaattcgt
     4201 tcttgttgtt tttggggaag tctttgtatt caacctggaa tccctggact ctggaaagat
     4261 agtccagttg ctcagagggt ctcgtgagag gtccatgggg tacgtggcct gaagagatgt
     4321 tattctttaa aatggtctcg gctgtgggcg aggtgccccc atacaacaac tctcgggcta
     4381 tcatggcagt taccgtggcc ttggcaggat tcggagctgc cctggtaaaa tctttggtgt
     4441 gatgtccttg actaactcct acagcctggg cgacctcggg caccatggga agaattccag
     4501 caggcagctg ctgatgactt agataaggca tcctgaactc atcctcttta ttactagtcc
     4561 cattttcatc cccagagcca ggttcaaaaa aggttacttt tcttccatcc cctggtttct
     4621 ttatgggtgt cttctcctct gacttgagtg cgggtttggt gggctgcgcc tgcgggactt
     4681 tgaaaccaag gatctccagc atgttctcgg ctgcattgtg cttggccacc ttcttgttgg
     4741 tgcccgttcc ttctgcagtg tggtttccaa ccttcacctg catcacaaac tccctgcggc
     4801 gcgggaggcc tcgctctgtg aggagcgtgt actctggctc cttctccttt tttgcctgct
     4861 ggatctgggc cagtcggcta atcggattga tcccctggcc atattctggg cttgtctgtg
     4921 gcttgactat gggttttgtt ttctttttga ttctaggctt tactcgttca actgcaggca
     4981 ggggcggtaa cttcttcagc tcctcaagaa cagctatggc ggcatttttc tttgaaatct
     5041 tcttgctttt cccttcacct tcccccacaa actccccaac cgaaaccttg gtcacaaagt
     5101 tcttcatgtg gggtgggcca ctctcccggg ccacctcgaa attcacaggc aagttccgtt
     5161 taagtgcaat ctcaaacact tgacttattt cagatttatt gagattttct tcttcggatt
     5221 ctcttccatt cacctccagc ctctctggca ggggctcatt ctgcaggatc ctcaacgctt
     5281 tggcagcagc atcgtgtttc gcagcctgtc ttgtctttcc tttgccatta aattgctgtc
     5341 ctcccacaga aagttccact tgataaagta aaggtggaac tggaaatggg taaaagtacc
     5401 tcgggggata agcacctcct ctcatgttgt agttataggt ggactgcatc cgagagtaag
     5461 ggtcaacagg cttatacatt ggtttttttc caagtttcat gcacagtgca tttagttcta
     5521 cagtaggggt tatgctttct gcagctgcac tggtggatgt aatagatgca gagggtaaag
     5581 cagagttttg aatgggtcta cctgcatttt cagagggcag agagctagta gtagaaggaa
     5641 tactcatcaa aggctgtgag agaagagact ggttcttgtt cagtatttgg ctccctgaga
     5701 gagcagcaga tgggttctga acttgcactt gaacttgaga catctcgagt cgacccggga
     5761 attcggatcc aagctttgga ggagcgtaat ctggaacatc gtatgggtaa gccatatgtg
     5821 gaggagatcc accagatcca cctccacctc cgtagatgcc ggagttggcc gcgatggccg
     5881 agggaatcgg gtttccatct tttaggagac cttgcattgc cttaacaata agctcgcagt
     5941 cggaattcgt ggcgaaaatt ggaatggtta gttccatatt taagtaagat ctccatccgg
     6001 ctacaggaag ctcttcacca ccaacagtct gggttgccac tttaggcacc tcgactttga
     6061 tggtgtattt gcgattctgc gcagagctct gacgaacgct acaggttact ttgtaagcct
     6121 gtgatcgcga gttagagctg atccattcag cgaccccgtt agcgaagttg cttggggcga
     6181 cagtcacgtc gccagttccg ccattgtcga cgagaacgaa ctgagtaaag ttagaagcca
     6241 tggtggcaag cttgggtctc cctatagtga gtcgtattaa tttcgataag ccagtaagca
     6301 gtgggttctc tagttagcca gagagctctg cttatataga cctcccaccg tacacgccta
     6361 ccgcccattt gcgtcaatgg ggcggagttg ttacgacatt ttggaaagtc ccgttgattt
     6421 tggtgccaaa acaaactccc attgacgtca atggggtgga gacttggaaa tccccgtgag
     6481 tcaaaccgct atccacgccc attgatgtac tgccaaaacc gcatcaccat ggtaatagcg
     6541 atgactaata cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca
     6601 taatgccagg cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga
     6661 tacacttgat gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa
     6721 tggaaagtcc ctattggcgt tactatggga acatacgtca ttattgacgt caatgggcgg
     6781 gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac gcggaactcc
     6841 atatatgggc tatgaactaa tgaccccgta attgattact attaataact agtcaataat
     6901 caatgtcaac gcgtatatct ggcccgtaca tcgcgaagca gcgcaaaacg cctaacccta
     6961 agcagattct tcatgcaatt gtcggtcaag ccttgccttg ttgtagctta aattttgctc
     7021 gcgcactact cagcgacctc caacacacaa gcagggagca gatactggct taactatgcg
     7081 gcatcagagc agattgtact gagagtgcac cataggggat cgggagatct cccgatccgt
     7141 cgacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     7201 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     7261 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     7321 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     7381 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     7441 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     7501 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     7561 attctcagaa tgacttggtt gagtactcac cagtcacaga aa
//
