LOCUS       sequence-473831-        6948 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HshnRNPE2iso3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6948)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 6948)
  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-HshnRNPE2iso3_Z
FEATURES             Location/Qualifiers
     source          1..6948
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      895..904
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1333..1359
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(2501..2519)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2502..2519)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2539..2556)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2545..2769
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2815..3243
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2902..2921)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3112..3133
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3252..3272)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3257..3586
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3437..3572
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3499..3518
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3653..4447
                     /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(3707..3726)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4317..4336
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4621..4742
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4658..4677)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4712..4731
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4791..4807)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4791..4807)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4804..4826)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4815..4831
                     /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(4839..4845,4846..4863,4864..4869))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4884..4905
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5022..5039)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5193..5781)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5273..5292)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5952..6743,6744..6812))
                     /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     6575..6594
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6813..6917)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttgccacc
      901 atggcttcta actttactca gttcgttctc gtcgacaatg gcggaactgg cgacgtgact
      961 gtcgccccaa gcaacttcgc taacggggtc gctgaatgga tcagctctaa ctcgcgatca
     1021 caggcttaca aagtaacctg tagcgttcgt cagagctctg cgcagaatcg caaatacacc
     1081 atcaaagtcg aggtgcctaa agtggcaacc cagactgttg gtggtgaaga gcttcctgta
     1141 gccggatgga gatcttactt aaatatggaa ctaaccattc caattttcgc cacgaattcc
     1201 gactgcgagc ttattgttaa ggcaatgcaa ggtctcctaa aagatggaaa cccgattccc
     1261 tcggccatcg cggccaactc cggcatctac ggaggtggag gtggatctgg tggatctcct
     1321 ccacatatgg cttacccata cgatgttcca gattacgctc ctccaaagct tggatccatg
     1381 gacaccggtg tgattgaagg tggattaaat gtcactctca ccatccggct acttatgcat
     1441 ggaaaggaag ttggcagtat catcggaaag aaaggagaat cagttaagaa gatgcgcgag
     1501 gagagtggtg cacgtatcaa catctcagaa gggaattgtc ctgagagaat tatcactttg
     1561 gctggaccca ctaatgccat cttcaaagcc tttgctatga tcattgacaa actggaagag
     1621 gacataagca gctctatgac caatagcaca gctgccagta gacccccggt caccctgagg
     1681 ctggtggtcc ctgctagtca gtgtggctct ctcattggaa aaggtggatg caagatcaag
     1741 gaaatacgag agagtacagg ggctcaggtc caggtggcag gggatatgct acccaactca
     1801 actgagcggg ccatcactat tgctggcatt ccacaatcca tcattgagtg tgtcaaacag
     1861 atctgcgtgg tcatgttgga gtcccccccg aagggcgtga ccatcccgta ccggcccaag
     1921 ccgtccagct ctccggtcat ctttgcaggt ggtcaggaca ggtacagcac aggcagcgac
     1981 agtgcgagct ttccccacac caccccgtcc atgtgcctca accctgacct ggagggacca
     2041 cctctagagg cctataccat tcaaggacag tatgccattc cacagccaga ttcgaccaag
     2101 ctgcaccagt tggcaatgca acagtctcat tttcccatga cgcatggcaa caccggattc
     2161 agtggcattg aatccagctc tccagaggtg aaaggctatt gggcaggttt ggatgcatct
     2221 gctcagacta cttctcatga actcaccatt ccaaacgatt tgattggctg cataatcggg
     2281 cgtcaaggcg ccaaaatcaa tgagatccgt cagatgtctg gggcgcagat caaaattgcg
     2341 aacccagtgg aaggatctac tgataggcag gttaccatca ctggatctgc tgccagcatt
     2401 agcctggctc aatatctaat caatgtcagg ctttcctcgg agacgggtgg catggggagc
     2461 agctagctcg agtctagagc ggccgcctag agggccctat tctatagtgt cacctaaatg
     2521 ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     2581 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     2641 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     2701 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     2761 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     2821 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     2881 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     2941 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     3001 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     3061 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     3121 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     3181 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     3241 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     3301 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     3361 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     3421 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     3481 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     3541 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     3601 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     3661 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     3721 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     3781 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     3841 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     3901 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     3961 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     4021 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     4081 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     4141 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     4201 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     4261 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     4321 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     4381 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     4441 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     4501 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     4561 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     4621 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     4681 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     4741 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     4801 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     4861 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     4921 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     4981 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     5041 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     5101 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     5161 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5221 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5281 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5341 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5401 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5461 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5521 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5581 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5641 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5701 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5761 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5821 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5881 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5941 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     6001 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     6061 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6121 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6181 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6241 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6301 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6361 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6421 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6481 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6541 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6601 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6661 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6721 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6781 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6841 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6901 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
