LOCUS       sequence-211048-        8877 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a 500-nt firefly luciferase circular RNA in Drosophila 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8877)
  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 8877)
  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: "Genes Dev. 
            2018 May 17. pii: gad.314856.118. doi: 10.1101/gad.314856.118."
COMMENT     Sequence Label: Hy_pMT dati MCS circfirefly_500
FEATURES             Location/Qualifiers
     source          1..8877
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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    2317..2438
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3148..3165)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3319..3907)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3399..3418)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4078..4869,4870..4938))
                     /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     4701..4720
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4939..5043)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5111..5129
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6525..6804
                     /label=copia promoter
                     /note="strong promoter from the Drosophila transposable 
                     element copia (Sinclair et al., 1986)"
     CDS             6850..7875
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             6850..7875
                     /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          8238..8303
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             8433..8453
                     /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 ctaccacccg gggagctcca ggtattgttt tgaagatttt
     1621 tctgcttatc tatatacttt aaagggaacc tggtaatcga atattcgatg cagtccaaac
     1681 aagagtagta aaaaatcaac agagaacgct attcgagttt ttagactaga ctatatgtac
     1741 acccgatatt tagctgttac gagcgcaaca atgaactttc aaaattgtgt gactgttttg
     1801 gacgttttgc aggtgtaagt aggggcgtgg ccaaagtgtt tttggtatgt caataaaaat
     1861 tggcaagaca atgaaaacga agattaatca aaacattttt tgtgggggcg tggcaaacag
     1921 ttttttggca aatcgatttt attttatcga tgaaataata tgaatacatt ttgcaagagc
     1981 gtgggagtgg cagttctgga cgctttgtgg gcgttagagg atgcgtagca acaaacttgc
     2041 gctgcgatct atgcttaatc tcaactttct agcttttagt tagatagttt ctgaaatctc
     2101 gacgttcata caggcggata tggccagttc gactcgtctt ttgatgctaa tcaagaatat
     2161 atgtagttta tatggtcgga aacgcttcct tctacctgtt acataccttt aaacaaatct
     2221 aatatacgct ttccctctac gagtaatggg tatgaaaaca aaaatcaaca cgcatcaacg
     2281 cattttggaa agttgaaggc gtgatagtgc ggccgcaact tgtttattgc agcttataat
     2341 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     2401 tctagttgtg gtttgtccaa actcatcaat gtatcttaac tagtggatcc actaggggcc
     2461 gccgacgcga ggctggatgg ccttccccat tatgattctt ctcgcttccg gcggcatcgg
     2521 gatgcccgcg ttgcaggcca tgctgtccag gcaggtagat gacgaccatc agggacagct
     2581 tcaaggatcg ctcgcggctc ttaccagcct aacttcgatc attggaccgc tgatcgtcac
     2641 ggcgatttat gccgcctcgg cgagcacatg gaacgggttg gcatggattg taggcgccgc
     2701 cctatacctt gtctgcctcc ccgcgttgcg tcgcggtgca tggagccggg ccacctcgac
     2761 ctgaatggaa gccggcggca cctcgctaac ggattcacca ctccaagaat tggagccaat
     2821 caattcttgc ggagaactgt gaatgcgcaa accaaccctt ggcagaacat atccatcgcg
     2881 tccgccatct ccagcagccg cacgcggcgc atctcgggca gcgttgggtc ctggccacgg
     2941 gtgcgcatga tcgtgctcct gtcgttgagg acccggctag gctggcgggg ttgccttact
     3001 ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg ctgcaaaacg
     3061 tctgcgacct gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta aagtctggaa
     3121 acgcggaagt cagcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     3181 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     3241 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3301 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3361 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3421 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3481 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3541 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3601 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3661 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3721 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     3781 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3841 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3901 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3961 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     4021 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     4081 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     4141 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     4201 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     4261 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4321 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4381 tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     4441 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     4501 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     4561 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4621 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4681 ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4741 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4801 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4861 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4921 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4981 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     5041 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     5101 aacctataaa aataggcgta tcacgaggcc ctttcgtctt caagaattca catttgtacg
     5161 aatttttttt ttatcaaaag ttcgagtttt tcaccaattt cctcatcaac cgagcaaggc
     5221 aaacggcttt gaataatatg gtgttatata tacatatatc aaatcgctgc tgactgcgtg
     5281 attgatggcc ccaagattac atattatcga atcaggattc agaaggagat caatgtcaaa
     5341 tgcggacagg aacatgaaag acgcctgtta tgcgcaatta aaaatttggg tttaattgct
     5401 gtggaaactg ttgttggcgg catcttaagt tcctgtttaa caacatcaac tacttatgta
     5461 cgtagaagcg tttaagccat ttgcatacag atgagaactg gcttttgtgc taatcagtca
     5521 agatgactcc gatgatgatg actcattacc tgaccagttt tcgctgcttt cttttcaaca
     5581 actacttgta tatgtattgt atccaatagc aatacattga atttccatgg tctagtcacg
     5641 tattatcatt taattgacac caagtcgtgt tattgttgag ctatcgagtt cagctcaaac
     5701 atttcttatt cccatgaata agccggcaaa aatatgcaat ctatgaaagt taatataagc
     5761 aaaccttact ttgactcaat accaatgcac tttgtgtcga taggttcacg caattgaggc
     5821 gattattccg ataacccaag cgattgactg ttcccgtttc gattccaatt gaaatttgga
     5881 aatgtacaat agttttgcta tatgctgtca agtacgctct tatcttctct gggttttctt
     5941 cagagtttcg aaacgcttct tctttttttt gttttttttt ttttggaatc tcgtattttg
     6001 gaaggggctc ccctctggaa tttgttacac tgtcgttatc attgcgaaca agcggcccga
     6061 agctatcagc gactttaaca tttacaatgc acttttttac gaccaattaa atgtacattt
     6121 tcctttcttc gcccgttgat aagcgaacgc gatgtggcgc aggcaatgtg ttgctcttgc
     6181 gacacaaacg caatcaaaat ggattcaatt tcgctttttc ccagtgaaac gaagaacgaa
     6241 ccgaccatca tgatatgctc ctctgcatgt tgcgtattga atcaatgaca atttcaatta
     6301 agccgcccgt tcgtcatgcg ttttcgtgcg cttcgaaatg ctgataacgc tgctgtcctc
     6361 caactgcttt gcatgtggac acaattccat ttatttaatt cttttatttg gatcggttaa
     6421 attaaaaagc gccttgttac gcatttaacg ttgtttccgg tgcgtggtgg tttcatgctt
     6481 ctgggaacgg caaatgggtt taggattggg aacccctcat catctgttgg aatatactat
     6541 tcaacctaca aaagtaacgt taaacaacac tactttatat ttgatatgaa tggccacacc
     6601 ttttatgcca taaaacatat tgtaagagaa taccactctt tttattcctt ctttccttct
     6661 tgtacgtttt ttgctgtaag taggtcgtgg tgctggtgtt gcagttgaaa taacttaaaa
     6721 tataaatcat aaaactcaaa cataaacttg actatttatt tatttattaa gaaaggaaat
     6781 ataaattata aattacaaca ggttatggac ctgcagccaa gcttggcgct cgtccggggg
     6841 caatgagata tgaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa
     6901 aagttcgaca gcgtctccga cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc
     6961 agcttcgatg taggagggcg tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc
     7021 tacaaagatc gttatgttta tcggcacttt gcatcggccg cgctcccgat tccggaagtg
     7081 cttgacattg gggaattcag cgagagcctg acctattgca tctcccgccg tgcacagggt
     7141 gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag
     7201 gccatggatg cgatcgctgc ggccgatctt agccagacga gcgggttcgg cccattcgga
     7261 ccgcaaggaa tcggtcaata cactacatgg cgtgatttca tatgcgcgat tgctgatccc
     7321 catgtgtatc actggcaaac tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct
     7381 ctcgatgagc tgatgctttg ggccgaggac tgccccgaag tccggcacct cgtgcacgcg
     7441 gatttcggct ccaacaatgt cctgacggac aatggccgca taacagcggt cattgactgg
     7501 agcgaggcga tgttcgggga ttcccaatac gaggtcgcca acatcttctt ctggaggccg
     7561 tggttggctt gtatggagca gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca
     7621 ggatcgccgc ggctccgggc gtatatgctc cgcattggtc ttgaccaact ctatcagagc
     7681 ttggttgacg gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc
     7741 cgatccggag ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg
     7801 accgatggct gtgtagaagt actcgccgat agtggaaacc gacgccccag cactcgtccg
     7861 agggcaaagg aatagagtag atgccgaccg aacaagagct gatttcgaga acgcctcagc
     7921 cagcaactcg cgcgagccta gcaaggcaaa tgcgagagaa cggccttacg cttggtggca
     7981 cagttctcgt ccacagttcg ctaagctcgc tcggctgggt cgcgggaggg ccggtcgcag
     8041 tgattcaggc ccttctggat tgtgttggtc cccagggcac gattgtcatg cccacgcact
     8101 cgggtgatct gactgatccc gcagattgga gatcgccgcc cgtgcctgcc gattgggtgc
     8161 agatctttgt gaaggaacct tacttctgtg gtgtgacata attggacaaa ctacctacag
     8221 agatttaaag ctctaaggta aatataaaat ttttaagtgt ataatgtgtt aaactactga
     8281 ttctaattgt ttgtgtattt tagattccaa cctatggaac tgatgaatgg gagcagtggt
     8341 ggaatgcctt taatgaggaa aacctgtttt gctcagaaga aatgccatct agtgatgatg
     8401 aggctactgc tgactctcaa cattctactc ctccaaaaaa gaagagaaag gtagaagacc
     8461 ccaaggactt tccttcagaa ttgctaagtt ttttgagtca tgctgtgttt agtaatagaa
     8521 ctcttgcttg ctttgctatt tacaccacaa aggaaaaagc tgcactgcta tacaagaaaa
     8581 ttatggaaaa atattctgta acctttataa gtaggcataa cagttataat cataacatac
     8641 tgttttttct tactccacac aggcatagag tgtctgctat taataactat gctcaaaaat
     8701 tgtgtacctt tagcttttta atttgtaaag gggttaataa ggaatatttg atgtatagtg
     8761 ccttgactag agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     8821 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgtt
//
