LOCUS       sequence-456520-        6960 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6960)
  AUTHORS   Hoell JI, Larsson E, Runge S, Nusbaum JD, Duggimpudi S, Farazi TA, 
            Hafner M, Borkhardt A, Sander C, Tuschl T
  TITLE     RNA targets of wild-type and mutant FET family proteins.
  JOURNAL   Nat Struct Mol Biol. 2011 Nov 13. doi: 10.1038/nsmb.2163.
  PUBMED    22081015
REFERENCE   2  (bases 1 to 6960)
  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: "Nat Struct 
            Mol Biol. 2011 Nov 13. doi: 10.1038/nsmb.2163."
COMMENT     Sequence Label: pDEST_TO_FUS
FEATURES             Location/Qualifiers
     source          1..6960
                     /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"
     CDS             993..1016
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1017..1043
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    1059..1083
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1490..1501
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     protein_bind    complement(2714..2738)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2791..2832
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(2912..2929)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2918..3142
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3275..3294)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3426..3473
                     /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             3481..4503
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3481..4503
                     /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    4633..4754
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4670..4689)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4724..4743
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4803..4819)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4803..4819)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4816..4838)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4827..4843
                     /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(4851..4857,4858..4875,4876..4881))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4896..4917
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5034..5051)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5205..5793)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5285..5304)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5964..6755,6756..6824))
                     /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     6587..6606
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6825..6929)
                     /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 cttggtacca tggactacaa ggacgacgat gacaagtacc
     1021 cttatgacgt gcccgattac gctttgaacc agatatcaac aagtttgtac aaaaaagcag
     1081 gctccaccat gggaaccaat tcagtcgaca tggcctcaaa cgattatacc caacaagcaa
     1141 cccaaagcta tggggcctac cccacccagc ccgggcaggg ctattcccag cagagcagtc
     1201 agccctacgg acagcagagt tacagtggtt atagccagtc cacggacact tcaggatatg
     1261 gccagagcag ctattcttct tatggccaga gccagaacac aggctatgga actcagtcaa
     1321 ctccccaggg atatggctcg actggcggct atggcagtag ccagagctcc caatcgtctt
     1381 acgggcagca gtcctcctac cctggctatg gccagcagcc agctcccagc agcacctcgg
     1441 gaagttacgg tagcagttct cagagcagca gctatgggca gccccagagt gggagctaca
     1501 gccagcagcc tagctatggt ggacagcagc aaagctatgg acagcagcaa agctataatc
     1561 cccctcaggg ctatggacag cagaaccagt acaacagcag cagtggtggt ggaggtggag
     1621 gtggaggtgg aggtaactat ggccaagatc aatcctccat gagtagtggt ggtggcagtg
     1681 gtggcggtta tggcaatcaa gaccagagtg gtggaggtgg cagcggtggc tatggacagc
     1741 aggaccgtgg aggccgcggc aggggtggca gtggtggcgg cggcggcggc ggcggtggtg
     1801 gttacaaccg cagcagtggt ggctatgaac ccagaggtcg tggaggtggc cgtggaggca
     1861 gaggtggcat gggcggaagt gaccgtggtg gcttcaataa atttggtggc cctcgggacc
     1921 aaggatcacg tcatgactcc gaacaggata attcagacaa caacaccatc tttgtgcaag
     1981 gcctgggtga gaatgttaca attgagtctg tggctgatta cttcaagcag attggtatta
     2041 ttaagacaaa caagaaaacg ggacagccca tgattaattt gtacacagac agggaaactg
     2101 gcaagctgaa gggagaggca acggtctctt ttgatgaccc accttcagct aaagcagcta
     2161 ttgactggtt tgatggtaaa gaattctccg gaaatcctat caaggtctca tttgctactc
     2221 gccgggcaga ctttaatcgg ggtggtggca atggtcgtgg aggccgaggg cgaggaggac
     2281 ccatgggccg tggaggctat ggaggtggtg gcagtggtgg tggtggccga ggaggatttc
     2341 ccagtggagg tggtggcggt ggaggacagc agcgagctgg tgactggaag tgtcctaatc
     2401 ccacctgtga gaatatgaac ttctcttgga ggaatgaatg caaccagtgt aaggccccta
     2461 aaccagatgg cccaggaggg ggaccaggtg gctctcacat ggggggtaac tacggggatg
     2521 atcgtcgtgg tggcagagga ggctatgatc gaggcggcta ccggggccgc ggcggggacc
     2581 gtggaggctt ccgagggggc cggggtggtg gggacagagg tggctttggc cctggcaaga
     2641 tggattccag gggtgagcac agacaggatc gcagggagag gccgtattaa gcggccgcac
     2701 tcgagatatc tagacccagc tttcttgtac aaagtggttg atatccagca cagtggcggc
     2761 cgctcaagtc tagagggccc gcggttcgaa ggtaagccta tccctaaccc tctcctcggt
     2821 ctcgattcta cgcgtacctg atccgcggcc gcatagataa ctgatccagt gtgcctcgag
     2881 tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     2941 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     3001 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     3061 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     3121 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     3181 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3241 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     3301 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     3361 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     3421 acctagaagt tcctattccg aagttcctat tctctagaaa gtataggaac ttccttggcc
     3481 aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc
     3541 gtctccgacc tgatgcagct ctcggagggc gaagaatctc gtgctttcag cttcgatgta
     3601 ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg atggtttcta caaagatcgt
     3661 tatgtttatc ggcactttgc atcggccgcg ctcccgattc cggaagtgct tgacattggg
     3721 gaattcagcg agagcctgac ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa
     3781 gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc catggatgcg
     3841 atcgctgcgg ccgatcttag ccagacgagc gggttcggcc cattcggacc gcaaggaatc
     3901 ggtcaataca ctacatggcg tgatttcata tgcgcgattg ctgatcccca tgtgtatcac
     3961 tggcaaactg tgatggacga caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg
     4021 atgctttggg ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc
     4081 aacaatgtcc tgacggacaa tggccgcata acagcggtca ttgactggag cgaggcgatg
     4141 ttcggggatt cccaatacga ggtcgccaac atcttcttct ggaggccgtg gttggcttgt
     4201 atggagcagc agacgcgcta cttcgagcgg aggcatccgg agcttgcagg atcgccgcgg
     4261 ctccgggcgt atatgctccg cattggtctt gaccaactct atcagagctt ggttgacggc
     4321 aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg atccggagcc
     4381 gggactgtcg ggcgtacaca aatcgcccgc agaagcgcgg ccgtctggac cgatggctgt
     4441 gtagaagtac tcgccgatag tggaaaccga cgccccagca ctcgtccgag ggcaaaggaa
     4501 tagcacgtac tacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     4561 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     4621 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     4681 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     4741 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     4801 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     4861 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     4921 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     4981 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     5041 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     5101 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     5161 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5221 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5281 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5341 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     5401 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5461 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5521 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5581 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5641 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5701 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     5761 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     5821 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     5881 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     5941 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     6001 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     6061 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     6121 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     6181 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     6241 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6301 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6361 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6421 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6481 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6541 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6601 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6661 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     6721 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     6781 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     6841 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     6901 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
