LOCUS       sequence-205780-        6994 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  open reading frame of U1-70K inserted into pcDNA5-FRT-TetO-fSNAPc 
            using the KpnI and NotI restriction sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6994)
  AUTHORS   Braun JE, Friedman LJ, Gelles J, Moore MJ
  TITLE     Synergistic assembly of human pre-spliceosomes across introns and 
            exons.
  JOURNAL   Elife. 2018 Jun 22;7. pii: 37751. doi: 10.7554/eLife.37751.
  PUBMED    29932423
REFERENCE   2  (bases 1 to 6994)
  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: "Elife."; 
            date: "2018-06-22"; volume: "7. pii: 37751. doi"; pages: " 
            10.7554/eLife.37751"
COMMENT     Sequence Label: pcDNA5-FRT-TetO-U1-70K-fSNAPc
FEATURES             Location/Qualifiers
     source          1..6994
                     /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"
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     869..893
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      983..992
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2360..2905
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     primer_bind     complement(2946..2963)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2952..3176
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3309..3328)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3460..3507
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             3515..4537
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3515..4537
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    4667..4788
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4704..4723)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4758..4777
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4837..4853)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4837..4853)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4850..4872)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4861..4877
                     /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(4885..4891,4892..4909,4910..4915))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4930..4951
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5068..5085)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5239..5827)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5319..5338)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5998..6789,6790..6858))
                     /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     6621..6640
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6859..6963)
                     /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 ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag ctgccaccat ggacaagctt tccagcggta ccatgaccca
     1021 gttcctgccg cccaaccttc tggccctctt tgccccccgt gaccctattc catacctgcc
     1081 acccctggag aaactgccac atgaaaaaca ccacaatcaa ccttattgtg gcattgcacc
     1141 ttatatccgt gaatttgagg accctcgtga tgcccctcct ccaactcgtg ctgaaacccg
     1201 agaggagcgc atggagagga aaagacggga aaagattgag cggcgacagc aagaagtgga
     1261 gacagagctt aaaatgtggg accctcacaa tgatcccaat gctcaggggg atgccttcaa
     1321 gactctcttc gtggcgagag tgaattatga cacaacagaa tccaagctcc gtagggaatt
     1381 cgaagtctac ggacctatca aaagaataca catggtctac agtaagcggt caggaaagcc
     1441 ccgtggctat gccttcatcg agtacgaaca cgagcgagat atgcattcag catataagca
     1501 tgctgatggc aagaagattg atggcaggag ggtccttgtg gacgtggaga ggggccgaac
     1561 cgtgaagggc tggaggcccc ggcggctagg aggaggcctc ggtggtacaa gaagaggagg
     1621 ggctgatgtg aacatccggc attcaggccg cgatgacacc tcccgctacg atgagaggcc
     1681 cggcccctcc ccgcttccgc acagggaccg ggaccgggac cgtgagcggg agcgcagaga
     1741 gcggagccgg gagcgagaca aggagcgaga acggcgacgc tcccgctccc gggaccggcg
     1801 gaggcgctca cggagtcgcg acaaggagga gcggaggcgc tccagggagc ggagcaagga
     1861 caaggaccgg gaccggaagc ggcgaagcag ccggagtcgg gagcgggccc ggcgggagcg
     1921 ggagcgcaag gaggagctgc gtggcggcgg tggcgacatg gcggagccct ccgaggcggg
     1981 tgacgcgccc cctgatgatg ggcctccagg ggagctcggg cctgacggcc ctgacggtcc
     2041 agaggaaaag ggccgggatc gtgaccggga gcgacggcgg agccaccgga gcgagcgcga
     2101 gcggcgccgg gaccgggatc gtgaccgtga ccgtgaccgc gagcacaaac ggggggagcg
     2161 gggcagtgag cggggcaggg atgaggcccg aggtgggggc ggtggccagg acaacgggct
     2221 ggagggtctg ggcaacgaca gccgagacat gtacatggag tctgagggcg gcgacggcta
     2281 cctggctccg gagaatgggt atttgatgga ggctgcgccg gagtgcggcc gcggctccgg
     2341 aggctccggc gggagcggca tggacaaaga ctgcgaaatg aagcgcacca ccctggatag
     2401 ccctctgggc aagctggaac tgtctgggtg cgaacagggc ctgcaccgta tcatcttcct
     2461 gggcaaagga acatctgccg ccgacgccgt ggaagtgcct gccccagccg ccgtgctggg
     2521 cggaccagag ccactgatgc aggccaccgc ctggctcaac gcctactttc accagcctga
     2581 ggccatcgag gagttccctg tgccagccct gcaccaccca gtgttccagc aggagagctt
     2641 tacccgccag gtgctgtgga aactgctgaa agtggtgaag ttcggagagg tcatcagcta
     2701 cagccacctg gccgccctgg ccggcaatcc cgccgccacc gccgccgtga aaaccgccct
     2761 gagcggaaat cccgtgccca ttctgatccc ctgccaccgg gtggtgcagg gcgacctgga
     2821 cgtggggggc tacgagggcg ggctcgccgt gaaagagtgg ctgctggccc acgagggcca
     2881 cagactgggc aagcctgggc tgggttagct cgagtctaga gggcccgttt aaacccgctg
     2941 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     3001 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     3061 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     3121 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     3181 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     3241 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     3301 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3361 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     3421 ccccaaaaaa cttgattagg gtgatggttc acgtacctag aagttcctat tccgaagttc
     3481 ctattctcta gaaagtatag gaacttcctt ggccaaaaag cctgaactca ccgcgacgtc
     3541 tgtcgagaag tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctcgga
     3601 gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt
     3661 aaatagctgc gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatcggc
     3721 cgcgctcccg attccggaag tgcttgacat tggggaattc agcgagagcc tgacctattg
     3781 catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc
     3841 tgttctgcag ccggtcgcgg aggccatgga tgcgatcgct gcggccgatc ttagccagac
     3901 gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt
     3961 catatgcgcg attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt
     4021 cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga
     4081 agtccggcac ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg
     4141 cataacagcg gtcattgact ggagcgaggc gatgttcggg gattcccaat acgaggtcgc
     4201 caacatcttc ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga
     4261 gcggaggcat ccggagcttg caggatcgcc gcggctccgg gcgtatatgc tccgcattgg
     4321 tcttgaccaa ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca
     4381 gggtcgatgc gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc
     4441 ccgcagaagc gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa
     4501 ccgacgcccc agcactcgtc cgagggcaaa ggaatagcac gtactacgag atttcgattc
     4561 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     4621 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     4681 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4741 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     4801 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     4861 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     4921 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     4981 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5041 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5101 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5161 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5221 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5281 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5341 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5401 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     5461 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5521 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5581 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5641 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     5701 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     5761 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     5821 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     5881 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     5941 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6001 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6061 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6121 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6181 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6241 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6301 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6361 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6421 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     6481 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     6541 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     6601 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     6661 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     6721 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     6781 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     6841 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     6901 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     6961 gcgcacattt ccccgaaaag tgccacctga cgtc
//
