LOCUS       Exported                8514 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Expresses Flag-tagged human UAP56 (DDX39B) with 4 mutations that 
            impact circRNA nuclear export activity.
ACCESSION   .
VERSION     .
KEYWORDS    Hy_pMT Flag-UAP56 MUT
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8514)
  AUTHORS   Huang C, Liang D, Tatomer DC, Wilusz JE
  TITLE     A length-dependent evolutionarily conserved pathway controls nuclear
            export of circular RNAs.
  JOURNAL   Genes Dev. 2018 May 17. pii: gad.314856.118. doi: 
            10.1101/gad.314856.118.
  PUBMED    29773557
REFERENCE   2  (bases 1 to 8514)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8514
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..427
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     CDS             446..511
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1819..1940
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2650..2667)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2821..3409)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2901..2920)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3580..4440)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4203..4222
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4441..4545)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4613..4631
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6027..6306
                     /label=copia promoter
                     /note="strong promoter from the Drosophila transposable 
                     element copia (Sinclair et al., 1986)"
     CDS             6352..7377
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     intron          7740..7805
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             7935..7955
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    8380..8514
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 aattcgttgc aggacaggat gtggtgcccg atgtgactag ctctttgctg caggccgtcc
       61 tatcctctgg ttccgataag agacccagaa ctccggcccc ccaccgccca ccgccacccc
      121 catacatatg tggtacgcaa gtaagagtgc ctgcgcatgc cccatgtgcc ccaccaagag
      181 ctttgcatcc catacaagtc cccaaagtgg agaaccgaac caattcttcg cgggcagaac
      241 aaaagcttct gcacacgtct ccactcgaat ttggagccgg ccggcgtgtg caaaagaggt
      301 gaatcgaacg aaagacccgt gtgtaaagcc gcgtttccaa aatgtataaa accgagagca
      361 tctggccaat gtgcatcagt tgtggtcagc agcaaaatca agtgaatcat ctcagtgcaa
      421 ctaaaggggg gatctcgaga ccatggacta caaagaccat gacggtgatt ataaagatca
      481 tgacatcgat tacaaggatg acgatgacaa gaccggtggt accatggcag agaacgatgt
      541 ggacaatgag ctcttggact atgaagatga tgaggtggag acagcagctg ggggagatgg
      601 ggctgaggcc cctgccaaga aggatgtcaa gggctcctat gtctccatcc acagctctgg
      661 ctttcgtgac ttcctgctca agccagagtt gctccgggcc attgtcgact gtggctttga
      721 gcatccgtca gaagtccagc atgagtgcat ccctcaggcc attctgggaa tggatgtcct
      781 gtgccaggcc aagtcgggca tgggaaagac agcagtgttt gtcttggcca cactgcaaca
      841 gctggagcca gttactgggc aggtgtctgt gctggtgatg tgtcacactc gggagttggc
      901 ttttcagatc agcaaggaat atgagcgctt ctctaaatac atgcccaatg tcaaggttgc
      961 tgtttttttt ggtggtctgt ctatcaagaa ggatgaagag gtgctgaaga agaactgccc
     1021 gcatatcgtc gtggggactc caggccgtat cctagccctg gctcgaaata ggagcttcag
     1081 cctcaaacac attaaacact ttattttgga tgaatgtgat aagatgcttg aacagctcga
     1141 catgcgtcgg gatgtccagg aaatttttcg catgaccccc cacgagaagc aggtcatgat
     1201 gttcagtgct accttgagca aagagatccg tccagtctgc cgcaagttca tgcaagatcc
     1261 aatggagatc ttcgtggatg atgagacgaa gttgacgctg catgggttgc agcagtacta
     1321 cgtgaaactg aaggacaacg agaagaaccg gaagctcttt gaccttctgg atgtccttga
     1381 gttcaaccag gtggtgatct ttgtgaagtc tgtgcagcgg tgcattgcct tggcccagct
     1441 actagtggag cagaacttcc cagccattgc catccaccgt gggatgcccc aggaggagag
     1501 gctttctcgg tatcagcagt ttaaagattt tcaacgacga attcttgtgg ctaccaacct
     1561 atttggccga ggcatggaca tcgagcgggt gaacattgct tttaattatg acatgcctga
     1621 ggattctgac acctacctgc atcgggtggc cagagcaggc cggtttggca ccaagggctt
     1681 ggctatcaca tttgtgtccg atgagaatga tgccaagatc ctcaatgatg tgcaggatcg
     1741 ctttgaggtc aatattagtg agctgcctga tgagatagac atctcctcct acattgaaca
     1801 gacacggtag gcggccgcaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     1861 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     1921 aaactcatca atgtatctta actagtggat ccactagggg ccgccgacgc gaggctggat
     1981 ggccttcccc attatgattc ttctcgcttc cggcggcatc gggatgcccg cgttgcaggc
     2041 catgctgtcc aggcaggtag atgacgacca tcagggacag cttcaaggat cgctcgcggc
     2101 tcttaccagc ctaacttcga tcattggacc gctgatcgtc acggcgattt atgccgcctc
     2161 ggcgagcaca tggaacgggt tggcatggat tgtaggcgcc gccctatacc ttgtctgcct
     2221 ccccgcgttg cgtcgcggtg catggagccg ggccacctcg acctgaatgg aagccggcgg
     2281 cacctcgcta acggattcac cactccaaga attggagcca atcaattctt gcggagaact
     2341 gtgaatgcgc aaaccaaccc ttggaagaac atatccatcg cgtccgccat ctccagcagc
     2401 cgcacgcggc gcatctcggg cagcgttggg tcctggccac gggtgcgcat gatcgtgctc
     2461 ctgtcgttga ggacccggct aggctggcgg ggttgcctta ctggttagca gaatgaatca
     2521 ccgatacgcg agcgaacgtg aagcgactgc tgctgcaaaa cgtctgcgac ctgagcaaca
     2581 acatgaatgg tcttcggttt ccgtgtttcg taaagtctgg aaacgcggaa gtcagcgctc
     2641 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     2701 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     2761 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     2821 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     2881 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     2941 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3001 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3061 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3121 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3181 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3241 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     3301 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3361 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3421 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     3481 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     3541 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     3601 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     3661 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     3721 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     3781 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     3841 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctgcagg
     3901 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     3961 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     4021 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     4081 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     4141 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg
     4201 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     4261 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     4321 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     4381 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     4441 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     4501 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     4561 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     4621 tatcacgagg ccctttcgtc ttcaagaatt cacatttgta cgaatttttt ttttatcaaa
     4681 agttcgagtt tttcaccaat ttcctcatca accgagcaag gcaaacggct ttgaataata
     4741 tggtgttata tatacatata tcaaatcgct gctgactgcg tgattgatgg ccccaagatt
     4801 acatattatc gaatcaggat tcagaaggag atcaatgtca aatgcggaca ggaacatgaa
     4861 agacgcctgt tatgcgcaat taaaaatttg ggtttaattg ctgtggaaac tgttgttggc
     4921 ggcatcttaa gttcctgttt aacaacatca actacttatg tacgtagaag cgtttaagcc
     4981 atttgcatac agatgagaac tggcttttgt gctaatcagt caagatgact ccgatgatga
     5041 tgactcatta cctgaccagt tttcgctgct ttcttttcaa caactacttg tatatgtatt
     5101 gtatccaata gcaatacatt gaatttccat ggtctagtca cgtattatca tttaattgac
     5161 accaagtcgt gttattgttg agctatcgag ttcagctcaa acatttctta ttcccatgaa
     5221 taagccggca aaaatatgca atctatgaaa gttaatataa gcaaacctta ctttgactca
     5281 ataccaatgc actttgtgtc gataggttca cgcaattgag gcgattattc cgataaccca
     5341 agcgattgac tgttcccgtt tcgattccaa ttgaaatttg gaaatgtaca atagttttgc
     5401 tatatgctgt caagtacgct cttatcttct ctgggttttc ttcagagttt cgaaacgctt
     5461 cttctttttt ttgttttttt ttttttggaa tctcgtattt tggaaggggc tcccctctgg
     5521 aatttgttac actgtcgtta tcattgcgaa caagcggccc gaagctatca gcgactttaa
     5581 catttacaat gcactttttt acgaccaatt aaatgtacat tttcctttct tcgcccgttg
     5641 ataagcgaac gcgatgtggc gcaggcaatg tgttgctctt gcgacacaaa cgcaatcaaa
     5701 atggattcaa tttcgctttt tcccagtgaa acgaagaacg aaccgaccat catgatatgc
     5761 tcctctgcat gttgcgtatt gaatcaatga caatttcaat taagccgccc gttcgtcatg
     5821 cgttttcgtg cgcttcgaaa tgctgataac gctgctgtcc tccaactgct ttgcatgtgg
     5881 acacaattcc atttatttaa ttcttttatt tggatcggtt aaattaaaaa gcgccttgtt
     5941 acgcatttaa cgttgtttcc ggtgcgtggt ggtttcatgc ttctgggaac ggcaaatggg
     6001 tttaggattg ggaacccctc atcatctgtt ggaatatact attcaaccta caaaagtaac
     6061 gttaaacaac actactttat atttgatatg aatggccaca ccttttatgc cataaaacat
     6121 attgtaagag aataccactc tttttattcc ttctttcctt cttgtacgtt ttttgctgta
     6181 agtaggtcgt ggtgctggtg ttgcagttga aataacttaa aatataaatc ataaaactca
     6241 aacataaact tgactattta tttatttatt aagaaaggaa atataaatta taaattacaa
     6301 caggttatgg acctgcagcc aagcttggcg ctcgtccggg ggcaatgaga tatgaaaaag
     6361 cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg aaaagttcga cagcgtctcc
     6421 gacctgatgc agctctcgga gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg
     6481 cgtggatatg tcctgcgggt aaatagctgc gccgatggtt tctacaaaga tcgttatgtt
     6541 tatcggcact ttgcatcggc cgcgctcccg attccggaag tgcttgacat tggggaattc
     6601 agcgagagcc tgacctattg catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg
     6661 cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg aggccatgga tgcgatcgct
     6721 gcggccgatc ttagccagac gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa
     6781 tacactacat ggcgtgattt catatgcgcg attgctgatc cccatgtgta tcactggcaa
     6841 actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt
     6901 tgggccgagg actgccccga agtccggcac ctcgtgcacg cggatttcgg ctccaacaat
     6961 gtcctgacgg acaatggccg cataacagcg gtcattgact ggagcgaggc gatgttcggg
     7021 gattcccaat acgaggtcgc caacatcttc ttctggaggc cgtggttggc ttgtatggag
     7081 cagcagacgc gctacttcga gcggaggcat ccggagcttg caggatcgcc gcggctccgg
     7141 gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga gcttggttga cggcaatttc
     7201 gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg agccgggact
     7261 gtcgggcgta cacaaatcgc ccgcagaagc gcggccgtct ggaccgatgg ctgtgtagaa
     7321 gtactcgccg atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa ggaatagagt
     7381 agatgccgac cgaacaagag ctgatttcga gaacgcctca gccagcaact cgcgcgagcc
     7441 tagcaaggca aatgcgagag aacggcctta cgcttggtgg cacagttctc gtccacagtt
     7501 cgctaagctc gctcggctgg gtcgcgggag ggccggtcgc agtgattcag gcccttctgg
     7561 attgtgttgg tccccagggc acgattgtca tgcccacgca ctcgggtgat ctgactgatc
     7621 ccgcagattg gagatcgccg cccgtgcctg ccgattgggt gcagatcttt gtgaaggaac
     7681 cttacttctg tggtgtgaca taattggaca aactacctac agagatttaa agctctaagg
     7741 taaatataaa atttttaagt gtataatgtg ttaaactact gattctaatt gtttgtgtat
     7801 tttagattcc aacctatgga actgatgaat gggagcagtg gtggaatgcc tttaatgagg
     7861 aaaacctgtt ttgctcagaa gaaatgccat ctagtgatga tgaggctact gctgactctc
     7921 aacattctac tcctccaaaa aagaagagaa aggtagaaga ccccaaggac tttccttcag
     7981 aattgctaag ttttttgagt catgctgtgt ttagtaatag aactcttgct tgctttgcta
     8041 tttacaccac aaaggaaaaa gctgcactgc tatacaagaa aattatggaa aaatattctg
     8101 taacctttat aagtaggcat aacagttata atcataacat actgtttttt cttactccac
     8161 acaggcatag agtgtctgct attaataact atgctcaaaa attgtgtacc tttagctttt
     8221 taatttgtaa aggggttaat aaggaatatt tgatgtatag tgccttgact agagatcata
     8281 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     8341 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     8401 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     8461 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatc
//
