LOCUS       Exported                9077 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Expresses a 700-nt firefly luciferase circular RNA in Drosophila 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    Hy_pMT dati MCS circfirefly_700
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9077)
  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 9077)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9077
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     polyA_signal    1..135
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        136..562
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     polyA_signal    2517..2638
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3348..3365)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3519..4107)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3599..3618)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4278..5138)
                     /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     4901..4920
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5139..5243)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5311..5329
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6725..7004
                     /label=copia promoter
                     /note="strong promoter from the Drosophila transposable 
                     element copia (Sinclair et al., 1986)"
     CDS             7050..8075
                     /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          8438..8503
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             8633..8653
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
       61 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
      121 tatcatgtct ggatcaattc gttgcaggac aggatgtggt gcccgatgtg actagctctt
      181 tgctgcaggc cgtcctatcc tctggttccg ataagagacc cagaactccg gccccccacc
      241 gcccaccgcc acccccatac atatgtggta cgcaagtaag agtgcctgcg catgccccat
      301 gtgccccacc aagagctttg catcccatac aagtccccaa agtggagaac cgaaccaatt
      361 cttcgcgggc agaacaaaag cttctgcaca cgtctccact cgaatttgga gccggccggc
      421 gtgtgcaaaa gaggtgaatc gaacgaaaga cccgtgtgta aagccgcgtt tccaaaatgt
      481 ataaaaccga gagcatctgg ccaatgtgca tcagttgtgg tcagcagcaa aatcaagtga
      541 atcatctcag tgcaactaaa ggggggatct cgagcctctg ccatgactat cgtagagtaa
      601 aaagggttaa gcatttccga ccatataaag tataaatact cttgatcaga atcaatagcc
      661 gagtcgaact ggccatgtcc gcctcaacct atgaggattg aaacaagcat tcattctgaa
      721 tacatagacg cagcgcaagt ttgttgactc atattgtcac actcactcta acgcccacaa
      781 agcgccttaa cctgccacgg acaccttttt gaaattcgaa atatttcgtt atcttttcat
      841 atttttatta gttttgtaaa tttatatcga gttgcaaaaa aaactattcc acgccaacta
      901 taactcccac aaaccgccca aaaatgccac acccactctt ttaagaaatg attcaatttt
      961 gttggattaa ataagtcgtt ttccttattt caatctatat gccgaaaaca tattgaccac
     1021 gcccactctg acccctaaaa ccgcccaatt ttaaaaattc aaaaaaccct tttacgctta
     1081 acaacttttt ttgttttaca ggcaggtacc atggaagatg ccaaaaacat taagaagggc
     1141 ccagcgccat tctacccact cgaagacggg accgccggcg agcagctgca caaagccatg
     1201 aagcgctacg ccctggtgcc cggcaccatc gcctttaccg acgcacatat cgaggtggac
     1261 attacctacg ccgagtactt cgagatgagc gttcggctgg cagaagctat gaagcgctat
     1321 gggctgaata caaaccatcg gatcgtggtg tgcagcgaga atagcttgca gttcttcatg
     1381 cccgtgttgg gtgccctgtt catcggtgtg gctgtggccc cagctaacga catctacaac
     1441 gagcgcgagc tgctgaacag catgggcatc agccagccca ccgtcgtatt cgtgagcaag
     1501 aaagggctgc aaaagatcct caacgtgcaa aagaagctac cgatcataca aaagatcatc
     1561 atcatggata gcaagaccga ctaccagggc ttccaaagca tgtacacctt cgtgacttcc
     1621 catttgccac ccggcttcaa cgagtacgac ttcgtgcccg agagcttcga ccgggacaaa
     1681 accatcgccc tgatcatgaa cagtagtggc agtaccggat tgcccaaggg cgtagcccta
     1741 ccgcaccgca ccgcttgtgt ccgattcagt catgcccgcg accccacccg gggagctcca
     1801 ggtattgttt tgaagatttt tctgcttatc tatatacttt aaagggaacc tggtaatcga
     1861 atattcgatg cagtccaaac aagagtagta aaaaatcaac agagaacgct attcgagttt
     1921 ttagactaga ctatatgtac acccgatatt tagctgttac gagcgcaaca atgaactttc
     1981 aaaattgtgt gactgttttg gacgttttgc aggtgtaagt aggggcgtgg ccaaagtgtt
     2041 tttggtatgt caataaaaat tggcaagaca atgaaaacga agattaatca aaacattttt
     2101 tgtgggggcg tggcaaacag ttttttggca aatcgatttt attttatcga tgaaataata
     2161 tgaatacatt ttgcaagagc gtgggagtgg cagttctgga cgctttgtgg gcgttagagg
     2221 atgcgtagca acaaacttgc gctgcgatct atgcttaatc tcaactttct agcttttagt
     2281 tagatagttt ctgaaatctc gacgttcata caggcggata tggccagttc gactcgtctt
     2341 ttgatgctaa tcaagaatat atgtagttta tatggtcgga aacgcttcct tctacctgtt
     2401 acataccttt aaacaaatct aatatacgct ttccctctac gagtaatggg tatgaaaaca
     2461 aaaatcaaca cgcatcaacg cattttggaa agttgaaggc gtgatagtgc ggccgcaact
     2521 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     2581 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac
     2641 tagtggatcc actaggggcc gccgacgcga ggctggatgg ccttccccat tatgattctt
     2701 ctcgcttccg gcggcatcgg gatgcccgcg ttgcaggcca tgctgtccag gcaggtagat
     2761 gacgaccatc agggacagct tcaaggatcg ctcgcggctc ttaccagcct aacttcgatc
     2821 attggaccgc tgatcgtcac ggcgatttat gccgcctcgg cgagcacatg gaacgggttg
     2881 gcatggattg taggcgccgc cctatacctt gtctgcctcc ccgcgttgcg tcgcggtgca
     2941 tggagccggg ccacctcgac ctgaatggaa gccggcggca cctcgctaac ggattcacca
     3001 ctccaagaat tggagccaat caattcttgc ggagaactgt gaatgcgcaa accaaccctt
     3061 ggcagaacat atccatcgcg tccgccatct ccagcagccg cacgcggcgc atctcgggca
     3121 gcgttgggtc ctggccacgg gtgcgcatga tcgtgctcct gtcgttgagg acccggctag
     3181 gctggcgggg ttgccttact ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa
     3241 gcgactgctg ctgcaaaacg tctgcgacct gagcaacaac atgaatggtc ttcggtttcc
     3301 gtgtttcgta aagtctggaa acgcggaagt cagcgctctt ccgcttcctc gctcactgac
     3361 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     3421 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     3481 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3541 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     3601 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3661 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3721 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3781 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3841 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3901 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     3961 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4021 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     4081 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     4141 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     4201 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     4261 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     4321 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     4381 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     4441 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     4501 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     4561 gttaatagtt tgcgcaacgt tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg
     4621 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     4681 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     4741 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     4801 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     4861 atgcggcgac cgagttgctc ttgcccggcg tcaacacggg ataataccgc gccacatagc
     4921 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     4981 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     5041 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     5101 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     5161 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     5221 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     5281 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt
     5341 caagaactca catttgtacg aatttttttt ttatcaaaag ttcgagtttt tcaccaattt
     5401 cctcatcaac cgagcaaggc aaacggcttt gaataatatg gtgttatata tacatatatc
     5461 aaatcgctgc tgactgcgtg attgatggcc ccaagattac atattatcga atcaggattc
     5521 agaaggagat caatgtcaaa tgcggacagg aacatgaaag acgcctgtta tgcgcaatta
     5581 aaaatttggg tttaattgct gtggaaactg ttgttggcgg catcttaagt tcctgtttaa
     5641 caacatcaac tacttatgta cgtagaagcg tttaagccat ttgcatacag atgagaactg
     5701 gcttttgtgc taatcagtca agatgactcc gatgatgatg actcattacc tgaccagttt
     5761 tcgctgcttt cttttcaaca actacttgta tatgtattgt atccaatagc aatacattga
     5821 atttccatgg tctagtcacg tattatcatt taattgacac caagtcgtgt tattgttgag
     5881 ctatcgagtt cagctcaaac atttcttatt cccatgaata agccggcaaa aatatgcaat
     5941 ctatgaaagt taatataagc aaaccttact ttgactcaat accaatgcac tttgtgtcga
     6001 taggttcacg caattgaggc gattattccg ataacccaag cgattgactg ttcccgtttc
     6061 gattccaatt gaaatttgga aatgtacaat agttttgcta tatgctgtca agtacgctct
     6121 tatcttctct gggttttctt cagagtttcg aaacgcttct tctttttttt gttttttttt
     6181 ttttggaatc tcgtattttg gaaggggctc ccctctggaa tttgttacac tgtcgttatc
     6241 attgcgaaca agcggcccga agctatcagc gactttaaca tttacaatgc acttttttac
     6301 gaccaattaa atgtacattt tcctttcttc gcccgttgat aagcgaacgc gatgtggcgc
     6361 aggcaatgtg ttgctcttgc gacacaaacg caatcaaaat ggattcaatt tcgctttttc
     6421 ccagtgaaac gaagaacgaa ccgaccatca tgatatgctc ctctgcatgt tgcgtattga
     6481 atcaatgaca atttcaatta agccgcccgt tcgtcatgcg ttttcgtgcg cttcgaaatg
     6541 ctgataacgc tgctgtcctc caactgcttt gcatgtggac acaattccat ttatttaatt
     6601 cttttatttg gatcggttaa attaaaaagc gccttgttac gcatttaacg ttgtttccgg
     6661 tgcgtggtgg tttcatgctt ctgggaacgg caaatgggtt taggattggg aacccctcat
     6721 catctgttgg aatatactat tcaacctaca aaagtaacgt taaacaacac tactttatat
     6781 ttgatatgaa tggccacacc ttttatgcca taaaacatat tgtaagagaa taccactctt
     6841 tttattcctt ctttccttct tgtacgtttt ttgctgtaag taggtcgtgg tgctggtgtt
     6901 gcagttgaaa taacttaaaa tataaatcat aaaactcaaa cataaacttg actatttatt
     6961 tatttattaa gaaaggaaat ataaattata aattacaaca ggttatggac ctgcagccaa
     7021 gcttggcgct cgtccggggg caatgagata tgaaaaagcc tgaactcacc gcgacgtctg
     7081 tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga cctgatgcag ctctcggagg
     7141 gcgaagaatc tcgtgctttc agcttcgatg taggagggcg tggatatgtc ctgcgggtaa
     7201 atagctgcgc cgatggtttc tacaaagatc gttatgttta tcggcacttt gcatcggccg
     7261 cgctcccgat tccggaagtg cttgacattg gggaattcag cgagagcctg acctattgca
     7321 tctcccgccg tgcacagggt gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg
     7381 ttctgcagcc ggtcgcggag gccatggatg cgatcgctgc ggccgatctt agccagacga
     7441 gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata cactacatgg cgtgatttca
     7501 tatgcgcgat tgctgatccc catgtgtatc actggcaaac tgtgatggac gacaccgtca
     7561 gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac tgccccgaag
     7621 tccggcacct cgtgcacgcg gatttcggct ccaacaatgt cctgacggac aatggccgca
     7681 taacagcggt cattgactgg agcgaggcga tgttcgggga ttcccaatac gaggtcgcca
     7741 acatcttctt ctggaggccg tggttggctt gtatggagca gcagacgcgc tacttcgagc
     7801 ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc gtatatgctc cgcattggtc
     7861 ttgaccaact ctatcagagc ttggttgacg gcaatttcga tgatgcagct tgggcgcagg
     7921 gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca caaatcgccc
     7981 gcagaagcgc ggccgtctgg accgatggct gtgtagaagt actcgccgat agtggaaacc
     8041 gacgccccag cactcgtccg agggcaaagg aatagagtag atgccgaccg aacaagagct
     8101 gatttcgaga acgcctcagc cagcaactcg cgcgagccta gcaaggcaaa tgcgagagaa
     8161 cggccttacg cttggtggca cagttctcgt ccacagttcg ctaagctcgc tcggctgggt
     8221 cgcgggaggg ccggtcgcag tgattcaggc ccttctggat tgtgttggtc cccagggcac
     8281 gattgtcatg cccacgcact cgggtgatct gactgatccc gcagattgga gatcgccgcc
     8341 cgtgcctgcc gattgggtgc agatctttgt gaaggaacct tacttctgtg gtgtgacata
     8401 attggacaaa ctacctacag agatttaaag ctctaaggta aatataaaat ttttaagtgt
     8461 ataatgtgtt aaactactga ttctaattgt ttgtgtattt tagattccaa cctatggaac
     8521 tgatgaatgg gagcagtggt ggaatgcctt taatgaggaa aacctgtttt gctcagaaga
     8581 aatgccatct agtgatgatg aggctactgc tgactctcaa cattctactc ctccaaaaaa
     8641 gaagagaaag gtagaagacc ccaaggactt tccttcagaa ttgctaagtt ttttgagtca
     8701 tgctgtgttt agtaatagaa ctcttgcttg ctttgctatt tacaccacaa aggaaaaagc
     8761 tgcactgcta tacaagaaaa ttatggaaaa atattctgta acctttataa gtaggcataa
     8821 cagttataat cataacatac tgttttttct tactccacac aggcatagag tgtctgctat
     8881 taataactat gctcaaaaat tgtgtacctt tagcttttta atttgtaaag gggttaataa
     8941 ggaatatttg atgtatagtg ccttgactag agatcataat cagccatacc acatttgtag
     9001 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     9061 atgcaattgt tgttgtt
//
