LOCUS       sequence-119727-        8060 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human SNRNP70 fused to Flag-HA tag (N-terminus).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8060)
  AUTHORS   Beck DB, Narendra V, Drury WJ 3rd, Casey R, Jansen PW, Yuan ZF, 
            Garcia BA, Vermeulen M, Bonasio R
  TITLE     In Vivo Proximity Labeling for the Detection of Protein-Protein and 
            Protein-RNA Interactions.
  JOURNAL   J Proteome Res. 2014 Dec 5;13(12):6135-43. doi: 10.1021/pr500196b. 
            Epub 2014 Oct 29.
  PUBMED    25311790
REFERENCE   2  (bases 1 to 8060)
  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"; doi: "10.1021/pr500196b";
            journalName: "J Proteome Res"; date: "2014-12-5- 5"; volume: "13"; 
            issue: "12"; pages: "6135-43"
COMMENT     Sequence Label: pINTO-NFH::hSNRNP70
FEATURES             Location/Qualifiers
     source          1..8060
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(52..71)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1223..1602
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1603..1806
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1757..1777
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    1808..1826
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1829..1847
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1857..1881
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1964..1973
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1973..1996
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2006..2032
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2039..2059
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(3447..3464)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3453..3677
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3723..4151
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3810..3829)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4020..4041
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4160..4180)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4165..4494
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4345..4480
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4407..4426
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4542..4589
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4608..4982
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    5112..5233
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5149..5168)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5203..5222
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5282..5298)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5282..5298)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5295..5317)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5306..5322
                     /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(5330..5336,5337..5354,5355..5360))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5375..5396
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6145..6162)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6316..6901)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6396..6415)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7072..7863,7864..7932))
                     /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     7695..7714
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7933..8037)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg
       61 ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga
      121 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgttaatt
      181 aagctagcag tctgcaatga gttcgagttg gcgcgcctgt cattctaaat ctctctttca
      241 gcctaaagct ttttccccgt atccccccag gtgtctgcag gctcaaagag cagcgagaag
      301 cgttcagagg aaagcgatcc cgtgccacct tccccgtgcc cgggctgtcc ccgcacgctg
      361 ccggctcggg gatgcggggg gagcgccgga ccggagcgga gccccgggcg gctcgctgct
      421 gccccctagc gggggaggga cgtaattaca tccctggggg ctttgggagg gggctgtccc
      481 cgtgagctcc caggcgcgcc tgtcattcta aatctctctt tcagcctaaa gctttttccc
      541 cgtatccccc caggtgtctg caggctcaaa gagcagcgag aagcgttcag aggaaagcga
      601 tcccgtgcca ccttccccgt gcccgggctg tccccgcacg ctgccggctc ggggatgcgg
      661 ggggagcgcc ggaccggagc ggagccccgg gcggctcgct gctgccccct agcgggggag
      721 ggacgtaatt acatccctgg gggctttggg ggggggctgt ccccgtgagc tcccaggcgc
      781 gccaactcgg tactcgggga tgctagcgca ccatcaacga ggaggacatc ttggctaacc
      841 aacccctgtt gttacagtcc atccaggaca gacttggctc gctggtgggc caggacagcg
      901 ggtatgtggg gggtcttccc aagaacgtta aggaaaagct gctgagcttg aagactttac
      961 aaagcgagct attcgaagtt gaaaaggaat ttcaggtgga gatgtttgag ctagagaaca
     1021 agtttctgca gaagtacaag cccatttggg agcagcggtc caggatcatt tcaggacaag
     1081 agcaacccaa gccggaacag atcgcaaagg gccaagagat tgtggagtca ctcaatgaga
     1141 cggagttgca attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat
     1201 gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt
     1261 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     1321 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     1381 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     1441 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     1501 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     1561 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
     1621 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
     1681 gacgtcaatg ggagtttgtt ttggaaccaa aatcaacggg actttccaaa atgtcgtaac
     1741 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
     1801 agagctctcc ctatcagtga tagagatctc cctatcagtg atagagatcg tcgacgagct
     1861 cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga cctccataga
     1921 agacaccggg accgatccag cctccggact ctagcgttta aaggccacca tggattacaa
     1981 ggatgacgac gataagggct ccggataccc atacgatgtg ccagattacg ctggcagcga
     2041 gaacctgtac ttccagggct ccggaaccgg tagcggtacc gagctcggat ccatgaccca
     2101 gttcctgccg cccaaccttc tggccctctt tgccccccgt gaccctattc catacctgcc
     2161 acccctggag aaactgccac atgaaaaaca ccacaatcaa ccttattgtg gcattgcgcc
     2221 gtacattcga gagtttgagg acccccgaga tgcccctcct ccaactcgtg ctgaaacccg
     2281 agaggagcgc atggagagga aaagacggga aaagattgag cggcgacagc aagaagtgga
     2341 gacagagctt aaaatgtggg accctcacaa tgatcccaat gctcaggggg atgccttcaa
     2401 gactctcttc gtggcgagag tgaattatga cacaacagaa tccaagctcc ggagagagtt
     2461 tgaggtgtac ggacctatca aaagaataca catggtctac agtaagcggt caggaaagcc
     2521 ccgtggctat gccttcatcg agtacgaaca cgagcgagac atgcactccg cttacaaaca
     2581 cgcagatggc aagaagattg atggcaggag ggtccttgtg gacgtggaga ggggccgaac
     2641 cgtgaagggc tggaggcccc ggcggctagg aggaggcctc ggtggtacca gaagaggagg
     2701 ggctgatgtg aacatccggc attcaggccg cgatgacacc tcccgctacg atgagaggcc
     2761 cggcccctcc ccgcttccgc acagggaccg ggaccgggac cgtgagcggg agcgcagaga
     2821 gcggagccgg gagcgagaca aggagcgaga acggcgacgc tcccgctccc gggaccggcg
     2881 gaggcgctca cggagtcgcg acaaggagga gcggaggcgc tccagggagc ggagcaagga
     2941 caaggaccgg gaccggaagc ggcgaagcag ccggagtcgg gagcgggccc ggcgggagcg
     3001 ggagcgcaag gaggagctgc gtggtggcgg tggcgacatg gcggagccct ccgaggcggg
     3061 tgacgcgccc cctgatgatg ggcctccagg ggagctcggg cctgacggcc ctgacggtcc
     3121 agaggaaaag ggccgggatc gtgaccggga gcgacggcgg agccaccgga gcgagcgcga
     3181 gcggcgccgg gaccgggatc gtgaccgtga ccgtgaccgc gagcacaaac ggggggagcg
     3241 gggcagtgag cggggcaggg atgaggcccg aggtgggggc ggtggccagg acaacgggct
     3301 ggagggtctg ggcaacgaca gccgagacat gtacatggag tctgagggcg gcgacggcta
     3361 cctggctccg gagaatgggt atttgatgga ggctgcgccg gagtgatgac tcgagtctag
     3421 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     3481 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     3541 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     3601 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     3661 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     3721 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     3781 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     3841 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     3901 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     3961 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4021 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     4081 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     4141 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     4201 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     4261 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4321 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4381 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     4441 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     4501 ccgggagctt gtatatccat tttcggatct gatcagcacg tgttgacaat taatcatcgg
     4561 catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg gccaagttga
     4621 ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg
     4681 accggctcgg gttctcccgg gacttcgtgg aggacgactt cgccggtgtg gtccgggacg
     4741 acgtgaccct gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct
     4801 gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga
     4861 acttccggga cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg
     4921 agttcgccct gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggact
     4981 gacacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     5041 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     5101 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     5161 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     5221 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     5281 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     5341 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     5401 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     5461 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct agtctgcaat
     5521 gagttcgagt tggcgcgcct gtcattctaa atctctcttt cagcctaaag ctttttcccc
     5581 gtatcccccc aggtgtctgc aggctcaaag agcagcgaga agcgttcaga ggaaagcgat
     5641 cccgtgccac cttccccgtg cccgggctgt ccccgcacgc tgccggctcg gggatgcggg
     5701 gggagcgccg gaccggagcg gagccccggg cggctcgctg ctgcccccta gcgggggagg
     5761 gacgtaatta catccctggg ggctttggga gggggctgtc cccgtgagct cccaggcgcg
     5821 cctgtcattc taaatctctc tttcagccta aagctttttc cccgtatccc cccaggtgtc
     5881 tgcaggctca aagagcagcg agaagcgttc agaggaaagc gatcccgtgc caccttcccc
     5941 gtgcccgggc tgtccccgca cgctgccggc tcggggatgc ggggggagcg ccggaccgga
     6001 gcggagcccc gggcggctcg ctgctgcccc ctagcggggg agggacgtaa ttacatccct
     6061 gggggctttg ggggggggct gtccccgtga gctcccaggc gcgccaactc ggtactcggg
     6121 gatttaatta agctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     6181 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     6241 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     6301 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     6361 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     6421 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     6481 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     6541 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     6601 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     6661 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     6721 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     6781 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     6841 cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     6901 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6961 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     7021 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     7081 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     7141 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     7201 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     7261 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     7321 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     7381 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     7441 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     7501 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     7561 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     7621 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     7681 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     7741 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     7801 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     7861 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     7921 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     7981 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     8041 atttccccga aaagtgccac
//
