LOCUS       sequence-109669-        5681 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses V5- and SBP-tagged hnRNPK in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5681)
  TITLE     Izaurralde lab hnRNP
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5681)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pT7-V5-SBP-C1-HshnRNPK
FEATURES             Location/Qualifiers
     source          1..5681
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     613..632
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        613..631
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      659..668
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             668..709
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             731..844
                     /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"
     polyA_signal    2469..2590
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2506..2525)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2560..2579
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2597..3052)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2734..2755)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2946..2965
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3079..3183
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3185..3542
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3185..3376
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3208..3228)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3393..3528
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3577..4371
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3577..4371
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3631..3650)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4241..4260
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4559..4578)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4603..4650
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4979..5567
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5468..5487
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
      601 gcttatcgaa attaatacga ctcactatag ggagacccaa gctctagcgc taccggtcgc
      661 caccatgggt aagcctatcc ctaaccctct cctcggtctc gattctacgc ccggaagcgg
      721 tagcggtacg atggacgaga agaccaccgg ctgggggggc ggccacgtgg tggagggcct
      781 ggccggcgag ctggagcagc tgcgggccag gctggagcac caccctcagg gccagcggga
      841 gcccagatct cgagggatgg aaactgaaca gccagaagaa accttcccta acactgaaac
      901 caatggtgaa tttggtaaac gccctgcaga agatatggaa gaggaacaag catttaaaag
      961 atctagaaac actgatgaga tggttgaatt acgcattctg cttcagagca agaatgctgg
     1021 ggcagtgatt ggaaaaggag gcaagaatat taaggctctc cgtacagact acaatgccag
     1081 tgtttcagtc ccagacagca gtggccccga gcgcatattg agtatcagtg ctgatattga
     1141 aacaattgga gaaattctga agaaaatcat ccctaccttg gaagagggcc tgcagttgcc
     1201 atcacccact gcaaccagcc agctcccgct cgaatctgat gctgtggaat gcttaaatta
     1261 ccaacactat aaaggaagtg actttgactg cgagttgagg ctgttgattc atcagagtct
     1321 agcaggagga attattgggg tcaaaggtgc taaaatcaaa gaacttcgag agaacactca
     1381 aaccaccatc aagcttttcc aggaatgctg tcctcattcc actgacagag ttgttcttat
     1441 tggaggaaaa cccgataggg ttgtagagtg cataaagatc atccttgatc ttatatctga
     1501 gtctcccatc aaaggacgtg cacagcctta tgatcccaat ttttacgatg aaacctatga
     1561 ttatggtggt tttacaatga tgtttgatga ccgtcgcgga cgcccagtgg gatttcccat
     1621 gcggggaaga ggtggttttg acagaatgcc tcctggtcgg ggtgggcgtc ccatgcctcc
     1681 atctagaaga gattatgatg atatgagccc tcgtcgagga ccacctcccc ctcctcccgg
     1741 acgaggcggc cggggtggta gcagagctcg gaatcttcct cttcctccac caccaccacc
     1801 tagaggggga gacctcatgg cctatgacag aagagggaga cctggagacc gttacgacgg
     1861 catggttggt ttcagtgctg atgaaacttg ggactctgca atagatacat ggagcccatc
     1921 agaatggcag atggcttatg aaccacaggg tggctccgga tatgattatt cctatgcagg
     1981 gggtcgtggc tcatatggtg atcttggtgg acctattatt actacacaag taactattcc
     2041 caaagatttg gctggatcta ttattggcaa aggtggtcag cggattaaac aaatccgtca
     2101 tgagtcggga gcttcgatca aaattgatga gcctttagaa ggatccgaag atcggatcat
     2161 taccattaca ggaacacagg accagataca gaatgcacag tatttgctgc agaacagtgt
     2221 gaagcagtat gcagatgttg aaggattcta atgcaagata ttttttcttt tttatagtgt
     2281 gaagcagtat tctggaaagt ttttctaaga attctgcagt cgacggtacc gcgggcccgg
     2341 gatccaccgg atctagataa ctgatcataa tcagccatac cacatttgta gaggttttac
     2401 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     2461 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     2521 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     2581 atgtatctta acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     2641 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     2701 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     2761 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     2821 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     2881 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     2941 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     3001 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc
     3061 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3121 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3181 agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3241 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3301 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3361 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3421 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3481 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     3541 aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     3601 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     3661 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     3721 gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg
     3781 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     3841 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     3901 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     3961 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     4021 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     4081 gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat
     4141 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     4201 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     4261 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     4321 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     4381 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     4441 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     4501 tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg
     4561 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     4621 ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc
     4681 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt
     4741 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg
     4801 caggccctgc catagcctca ggttactcat atatacttta gattgattta aaacttcatt
     4861 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     4921 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     4981 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5041 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5101 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     5161 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5221 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5281 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5341 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5401 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5461 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5521 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     5581 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     5641 tatcccctga ttctgtggat aaccgtatta ccgccatgca t
//
