LOCUS       sequence-492411-        8345 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Flag-HA-hnRNPC in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8345)
  AUTHORS   Thomas A, Rehfeld F, Zhang H, Chang TC, Goodarzi M, Gillet F, 
            Mendell JT
  TITLE     RBM33 directs the nuclear export of transcripts containing GC-rich 
            elements.
  JOURNAL   Genes Dev. 2022 May 1;36(9-10):550-565. doi: 10.1101/gad.349456.122.
            Epub 2022 May 19.
  PUBMED    35589130
REFERENCE   2  (bases 1 to 8345)
  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: "Genes 
            Dev."; date: "2022-05-1"; volume: "36(9-10):550-565. doi"; pages: " 
            10.1101/gad.349456.122. Epub 2022 May 19"
COMMENT     Sequence Label: pLJM1-FH-hnRNPC
FEATURES             Location/Qualifiers
     source          1..8345
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    209..334
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    827..1060
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1245..1289
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     primer_bind     1587..1607
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     promoter        1896..2099
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2050..2070
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2176..2199
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2200..2226
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    3193..3310
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3359..3869
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     3788..3806
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             3891..4490
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3891..3910)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4387..4407
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4618..4851
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4923..5044
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4960..4979)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5014..5033
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5084..5219
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5239..5258)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5240..5258)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5268..5285)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5268..5284)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5277..5299)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5426..5881
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5513..5532)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5723..5744
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5907..6011
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6012..6080,6081..6872)
                     /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(6230..6249)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7043..7631
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7532..7551
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7785..7802
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7919..7940
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7955..7960,7961..7978,7979..7985)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7993..8009
                     /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     7998..8020
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8017..8033
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8017..8033
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8052..8072
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8054..8072
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        8100..8326
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             join(8327..8345,1..162)
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
ORIGIN
        1 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
       61 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
      121 gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga
      181 cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc
      241 gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg
      301 aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc
      361 gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat
      421 agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc
      481 agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga
      541 acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat
      601 aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaccac
      661 cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg gacaattgga
      721 gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca
      781 aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc
      841 ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac
      901 aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg
      961 aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa
     1021 tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg
     1081 gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg
     1141 aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa
     1201 gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat
     1261 tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt
     1321 ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg
     1381 ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc
     1441 acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa ggtggagaga
     1501 gagacagaga cagatccatt cgattagtga acggatctcg acggtatcga tcacgagact
     1561 agcctcgagc ggccgccccc ttcaccgagg gcctatttcc catgattcct tcatatttgc
     1621 atatacgata caaggctgtt agagagataa ttggaattaa tttgactgta aacacaaaga
     1681 tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa
     1741 aattatgttt taaaatggac tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     1801 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     1861 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
     1921 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
     1981 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
     2041 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg
     2101 gtttagtgaa ccgtcagatc cgctagcgct accgggtaca agtccggact cagatctcga
     2161 gctcaagctt cgatggacta caaggacgac gatgacaagt acccttatga cgtgcccgat
     2221 tacgctagcg gccgcatggc cagcaacgtt accaacaaga cagatcctcg ctccatgaac
     2281 tcccgtgtat tcattgggaa tctcaacact cttgtggtca agaaatctga tgtggaggca
     2341 atcttttcga agtatggcaa aattgtgggc tgctctgttc ataagggctt tgccttcgtt
     2401 cagtatgtta atgagagaaa tgcccgggct gctgtagcag gagaggatgg cagaatgatt
     2461 gctggccagg ttttagatat taacctggct gcagagccaa aagtgaaccg aggaaaagca
     2521 ggtgtgaaac gatctgcagc ggagatgtac gggtcagtaa cagaacaccc ttctccgtcc
     2581 cctctactca gctcctcttt tgacttggac tatgactttc aacgggacta ttatgatagg
     2641 atgtacagtt acccagcacg tgtacctcct cctcctccta ttgctcgggc tgtagtgccc
     2701 tcgaaacgtc agcgtgtatc aggaaacact tcacgaaggg gcaaaagtgg cttcaattct
     2761 aagagtggac agcggggatc ttccaagtct ggaaagttga aaggagatga ccttcaggcc
     2821 attaagaagg agctgaccca gataaaacaa aaagtggatt ctctcctgga aaacctggaa
     2881 aaaattgaaa aggaacagag caaacaagca gtagagatga agaatgataa gtcagaagag
     2941 gagcagagca gcagctccgt gaagaaagat gagactaatg tgaagatgga gtctgggggg
     3001 ggtgcagatg actctgctga ggagggggac ctactggatg atgatgataa tgaagatcgg
     3061 ggggatgacc agctggagtt gatcaaggat gatgaaaaag aggctgagga aggagaggat
     3121 gacagagaca gcgccaatgg cgaggatgac tcttaactcg acctcgagac aaatggcagt
     3181 attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat
     3241 agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat
     3301 tcaaaatttt cgggtttatt acagggacag cagagatcca ctttggccgc ggctcgaggg
     3361 ggttggggtt gcgccttttc caaggcagcc ctgggtttgc gcagggacgc ggctgctctg
     3421 ggcgtggttc cgggaaacgc agcggcgccg accctgggtc tcgcacattc ttcacgtccg
     3481 ttcgcagcgt cacccggatc ttcgccgcta cccttgtggg ccccccggcg acgcttcctg
     3541 ctccgcccct aagtcgggaa ggttccttgc ggttcgcggc gtgccggacg tgacaaacgg
     3601 aagccgcacg tctcactagt accctcgcag acggacagcg ccagggagca atggcagcgc
     3661 gccgaccgcg atgggctgtg gccaatagcg gctgctcagc agggcgcgcc gagagcagcg
     3721 gccgggaagg ggcggtgcgg gaggcggggt gtggggcggt agtgtgggcc ctgttcctgc
     3781 ccgcgcggtg ttccgcattc tgcaagcctc cggagcgcac gtcggcagtc ggctccctcg
     3841 ttgaccgaat caccgacctc tctccccagg gggatccacc ggagcttacc atgaccgagt
     3901 acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg
     3961 ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg
     4021 agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg
     4081 tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag
     4141 cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg
     4201 ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt
     4261 tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg
     4321 tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg
     4381 cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg
     4441 tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga cgcccgcccc
     4501 acgacccgca gcgcccgacc gaaaggagcg cacgacccca tgcatcggta cctttaagac
     4561 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     4621 aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct
     4681 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     4741 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     4801 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt
     4861 catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga
     4921 ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     4981 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     5041 cttatcatgt ctggctctag ctatcccgcc cctaactccg cccatcccgc ccctaactcc
     5101 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     5161 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     5221 agggacgtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt
     5281 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
     5341 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     5401 ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     5461 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     5521 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     5581 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     5641 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     5701 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     5761 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     5821 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     5881 taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     5941 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     6001 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     6061 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     6121 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     6181 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     6241 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     6301 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     6361 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     6421 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     6481 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     6541 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     6601 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     6661 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     6721 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     6781 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     6841 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     6901 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     6961 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     7021 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     7081 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     7141 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     7201 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     7261 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     7321 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     7381 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     7441 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     7501 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     7561 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     7621 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     7681 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     7741 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     7801 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     7861 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     7921 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     7981 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     8041 acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctg caagcttaat
     8101 gtagtcttat gcaatactct tgtagtcttg caacatggta acgatgagtt agcaacatgc
     8161 cttacaagga gagaaaaagc accgtgcatg ccgattggtg gaagtaaggt ggtacgatcg
     8221 tgccttatta ggaaggcaac agacgggtct gacatggatt ggacgaacca ctgaattgcc
     8281 gcattgcaga gatattgtat ttaagtgcct agctcgatac ataaacgggt ctctctggtt
     8341 agacc
//
