LOCUS       sequence-387277-        5730 bp    DNA     circular SYN 14-APR-2025
DEFINITION  expresses U2AF35 with N-terminal tags RFP and FKBP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5730)
  AUTHORS   Vester K, Preussner M, Holton N, Feng S, Schultz C, Heyd F, Wahl MC
  TITLE     Recruitment of a splicing factor to the nuclear lamina for its 
            inactivation.
  JOURNAL   Commun Biol. 2022 Jul 22;5(1):736. doi: 10.1038/s42003-022-03689-y.
  PUBMED    35869234
REFERENCE   2  (bases 1 to 5730)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Commun 
            Biol."; date: "2022-07-22"; volume: "5(1):736. doi"; pages: " 
            10.1038/s42003-022-03689-y"
COMMENT     Sequence Label: RFP-FKBP-U2AF35
FEATURES             Location/Qualifiers
     source          1..5730
                     /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"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             613..1287
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             613..1287
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(743..761)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1011..1030
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1205..1228
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             join(1345..1347,1348..1668)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     polyA_signal    2518..2639
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2555..2574)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2609..2628
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2646..3101)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2783..2804)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2995..3014
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3128..3232
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3234..3591
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3234..3425
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3257..3277)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3442..3577
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3626..4420
                     /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             3626..4420
                     /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(3680..3699)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4290..4309
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4608..4627)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4652..4699
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5028..5616
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5517..5536
                     /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 gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggcctc ctccgaggac gtcatcaagg agttcatgcg cttcaaggtg
      661 cgcatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc
      721 ccctacgagg gcacccagac cgccaagctg aaggtgacca agggcggccc cctgcccttc
      781 gcctgggaca tcctgtcccc tcagttccag tacggctcca aggcctacgt gaagcacccc
      841 gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg
      901 atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc
      961 gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg
     1021 cagaagaaga ccatgggctg ggaggcctcc accgagcgga tgtaccccga ggacggcgcc
     1081 ctgaagggcg agatcaagat gaggctgaag ctgaaggacg gcggccacta cgacgccgag
     1141 gtcaagacca cctacatggc caagaagccc gtgcagctgc ccggcgccta caagaccgac
     1201 atcaagctgg acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgagcgc
     1261 gccgagggcc gccactccac cggcgcctcc ggactcagat ctcgaagcgc ggccgcggga
     1321 gcaggaggag cagctcgagc ggcgatggga gtgcaggtgg aaaccatctc cccaggagac
     1381 gggcgcacct tccccaagcg cggccagacc tgcgtggtgc actacaccgg gatgcttgaa
     1441 gatggaaaga aatttgattc ctcccgggac agaaacaagc cctttaagtt tatgctaggc
     1501 aagcaggagg tgatccgagg ctgggaagaa ggggttgccc agatgagtgt gggtcagaga
     1561 gccaaactga ctatatctcc agattatgcc tatggtgcca ctgggcaccc aggcatcatc
     1621 ccaccacatg ccactctcgt cttcgatgtg gagcttctaa aactggaagg atccaccgga
     1681 tctagaatgg cggaatactt ggcctccatc ttcggcaccg aaaaagacaa agtcaactgt
     1741 tcattttatt tcaaaatcgg agcatgtcgt catggagaca gatgttctcg gttgcacaat
     1801 aaaccaacct ttagccagac cattgccctc ttgaacattt accgtaaccc tcaaaactct
     1861 tcccagtctg ctgacggttt gcgctgtgct gtgagcgacg tggagatgca ggagcactat
     1921 gatgagttct ttgaggaagt cttcactgag atggaagaga agtacgggga agtcgaggag
     1981 atgaacgtct gcgacaacct aggggaccac ctggtgggga acgtgtatgt caagtttcga
     2041 cgtgaggagg atgcagagaa agccgtgatc gacttgaata accgttggtt taatggacag
     2101 ccgatccatg cagagctgtc cccagtaact gacttcaggg aagcctgctg ccgccagtat
     2161 gaaatgggag agtgcacaag agggggcttc tgcaacttca tgcatttgaa gcccatctca
     2221 agagagctac gacgggagct gtatgggcgc cggcgcaaga agcatagatc caggtcccga
     2281 tccagggaac ggcgttcccg atccagagac cgtggacgcg gtggtggagg tggaggtgga
     2341 ggtggcggag gacgggagcg tgacaggagg cggtcaagag accgggagag atctggacga
     2401 ttcggataac tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     2461 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     2521 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     2581 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     2641 cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     2701 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     2761 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     2821 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     2881 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     2941 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     3001 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     3061 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg
     3121 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3181 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc
     3241 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3301 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3361 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3421 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3481 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     3541 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca
     3601 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     3661 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     3721 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     3781 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac
     3841 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     3901 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     3961 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     4021 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     4081 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     4141 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     4201 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     4261 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     4321 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     4381 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     4441 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     4501 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     4561 ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag
     4621 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     4681 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
     4741 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
     4801 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
     4861 atagcctcag gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     4921 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     4981 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     5041 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     5101 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     5161 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     5221 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     5281 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     5341 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     5401 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     5461 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     5521 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     5581 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     5641 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     5701 tctgtggata accgtattac cgccatgcat
//
