LOCUS       sequence-420976-        6816 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HshnRNPA1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6816)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 6816)
  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-HshnRNPA1_Z
FEATURES             Location/Qualifiers
     source          1..6816
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     319..338
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(557..661)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(736..755)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        927..1306
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1307..1510
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1461..1481
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1555..1574
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1555..1573
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1587..1596
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2025..2051
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2858..2869
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     promoter        complement(3061..3079)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3062..3079)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3099..3116)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3105..3329
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3375..3803
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3462..3481)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3672..3693
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3812..3832)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3817..4146
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3997..4132
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4059..4078
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4213..5007
                     /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(4267..4286)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4877..4896
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5181..5302
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5218..5237)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5272..5291
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5351..5367)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5351..5367)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5364..5386)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5375..5391
                     /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(5399..5405,5406..5423,5424..5429))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5444..5465
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5582..5599)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5753..6341)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5833..5852)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6512..6816,1..487,488..556))
                     /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"
ORIGIN
        1 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
       61 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
      121 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
      181 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
      241 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
      301 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
      361 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
      421 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
      481 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
      541 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
      601 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
      661 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct
      721 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc
      781 tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca
      841 aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg
      901 cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc
      961 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
     1021 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
     1081 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     1141 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
     1201 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
     1261 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
     1321 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
     1381 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
     1441 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
     1501 aagcagagct ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac
     1561 gactcactat agggagaccc aagcttgcca ccatggcttc taactttact cagttcgttc
     1621 tcgtcgacaa tggcggaact ggcgacgtga ctgtcgcccc aagcaacttc gctaacgggg
     1681 tcgctgaatg gatcagctct aactcgcgat cacaggctta caaagtaacc tgtagcgttc
     1741 gtcagagctc tgcgcagaat cgcaaataca ccatcaaagt cgaggtgcct aaagtggcaa
     1801 cccagactgt tggtggtgaa gagcttcctg tagccggatg gagatcttac ttaaatatgg
     1861 aactaaccat tccaattttc gccacgaatt ccgactgcga gcttattgtt aaggcaatgc
     1921 aaggtctcct aaaagatgga aacccgattc cctcggccat cgcggccaac tccggcatct
     1981 acggaggtgg aggtggatct ggtggatctc ctccacatat ggcttaccca tacgatgttc
     2041 cagattacgc tcctccaaag cttggatcca tgtctaagtc agagtctcct aaagagcccg
     2101 aacagctgag gaagctcttc attggagggt tgagctttga aacaactgat gagagcctga
     2161 ggagccattt tgagcaatgg ggaacgctca cggactgtgt ggtaatgaga gatccaaaca
     2221 ccaagcgctc caggggcttt gggtttgtca catatgccac tgtggaggag gtggatgcag
     2281 ctatgaatgc aaggccacac aaggtggatg gaagagttgt ggaaccaaag agagctgtct
     2341 ccagagaaga ttctcaaaga ccaggtgccc acttaactgt gaaaaagata tttgttggtg
     2401 gcattaaaga agacactgaa gaacatcacc taagagatta ttttgaacag tatggaaaaa
     2461 ttgaagtgat tgaagtcatg actgaccgag gcagtggcaa gaaaaggggc tttgcctttg
     2521 taacctttga cgaccatgac tccgtggata agattgtcat tcagaaatac catactgtga
     2581 atggccacaa ctgtgaagtt agaaaagccc tgtcaaagca agagatggct agtgcttcat
     2641 ccagccaaag aggtcgaagt ggttctggaa actttggtgg tggtcgtgga ggtggtttcg
     2701 gtgggaatga caacttcggt cgtggaggaa acttcagtgg tcgtggtggc tttggtggca
     2761 gccgtggtgg tggtggatat ggtggcagtg gggatggcta taatggattt ggtaatgatg
     2821 gaagcaattt tggaggtggt ggaagctaca atgattttgg gaattacaac aatcagtctt
     2881 caaattttgg acccatgaag ggaggaaatt ttggaggcag aagctctggc ccctatggcg
     2941 gtggaggcca atactttgca aaaccacgaa accaaggtgg ctatggcggt tccagcagca
     3001 gcagtagcta tggcagtggc agaagatttt aatctagagc ggccgcctag agggccctat
     3061 tctatagtgt cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc cttctagttg
     3121 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     3181 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     3241 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     3301 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     3361 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     3421 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     3481 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     3541 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     3601 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     3661 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     3721 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     3781 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     3841 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3901 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3961 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     4021 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     4081 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     4141 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     4201 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     4261 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     4321 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4381 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4441 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4501 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4561 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4621 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4681 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4741 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     4801 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     4861 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     4921 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     4981 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     5041 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     5101 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     5161 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     5221 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     5281 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     5341 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     5401 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     5461 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     5521 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     5581 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     5641 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     5701 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     5761 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     5821 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     5881 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     5941 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     6001 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     6061 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     6121 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     6181 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     6241 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     6301 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     6361 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     6421 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     6481 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     6541 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     6601 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     6661 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     6721 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     6781 taagtagttc gccagttaat agtttgcgca acgttg
//
