LOCUS       sequence-127583-        9377 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a GFP circular RNA in Drosophila.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9377)
  AUTHORS   Kramer MC, Liang D, Tatomer DC, Gold B, March ZM, Cherry S, Wilusz 
            JE
  TITLE     Combinatorial control of Drosophila circular RNA expression by 
            intronic repeats, hnRNPs, and SR proteins.
  JOURNAL   Genes Dev. 2015 Oct 8.
  PUBMED    26450910
REFERENCE   2  (bases 1 to 9377)
  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. 
            2015 Oct 8."
COMMENT     Sequence Label: Hy_pMT Laccase2 MCS-WT Split GFP
FEATURES             Location/Qualifiers
     source          1..9377
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(23..611)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(103..122)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(782..1573,1574..1642))
                     /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     1405..1424
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1643..1747)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1815..1833
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3229..3508
                     /label=copia promoter
                     /note="strong promoter from the Drosophila transposable 
                     element copia (Sinclair et al., 1986)"
     CDS             3554..4579
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             3554..4579
                     /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          4942..5007
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             5137..5157
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    5582..5716
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        5717..6143
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     polyA_signal    8398..8519
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9229..9246)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
       61 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
      121 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
      181 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
      241 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
      301 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
      361 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
      421 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
      481 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
      541 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
      601 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
      661 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
      721 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
      781 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
      841 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
      901 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
      961 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     1021 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     1081 acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     1141 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     1201 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     1261 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     1321 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     1381 gctcttgccc ggcgtcaaca cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     1441 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     1501 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     1561 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     1621 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     1681 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     1741 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     1801 cattaaccta taaaaatagg cgtatcacga ggccctttcg tcttcaagaa ttcacatttg
     1861 tacgaatttt ttttttatca aaagttcgag tttttcacca atttcctcat caaccgagca
     1921 aggcaaacgg ctttgaataa tatggtgtta tatatacata tatcaaatcg ctgctgactg
     1981 cgtgattgat ggccccaaga ttacatatta tcgaatcagg attcagaagg agatcaatgt
     2041 caaatgcgga caggaacatg aaagacgcct gttatgcgca attaaaaatt tgggtttaat
     2101 tgctgtggaa actgttgttg gcggcatctt aagttcctgt ttaacaacat caactactta
     2161 tgtacgtaga agcgtttaag ccatttgcat acagatgaga actggctttt gtgctaatca
     2221 gtcaagatga ctccgatgat gatgactcat tacctgacca gttttcgctg ctttcttttc
     2281 aacaactact tgtatatgta ttgtatccaa tagcaataca ttgaatttcc atggtctagt
     2341 cacgtattat catttaattg acaccaagtc gtgttattgt tgagctatcg agttcagctc
     2401 aaacatttct tattcccatg aataagccgg caaaaatatg caatctatga aagttaatat
     2461 aagcaaacct tactttgact caataccaat gcactttgtg tcgataggtt cacgcaattg
     2521 aggcgattat tccgataacc caagcgattg actgttcccg tttcgattcc aattgaaatt
     2581 tggaaatgta caatagtttt gctatatgct gtcaagtacg ctcttatctt ctctgggttt
     2641 tcttcagagt ttcgaaacgc ttcttctttt ttttgttttt ttttttttgg aatctcgtat
     2701 tttggaaggg gctcccctct ggaatttgtt acactgtcgt tatcattgcg aacaagcggc
     2761 ccgaagctat cagcgacttt aacatttaca atgcactttt ttacgaccaa ttaaatgtac
     2821 attttccttt cttcgcccgt tgataagcga acgcgatgtg gcgcaggcaa tgtgttgctc
     2881 ttgcgacaca aacgcaatca aaatggattc aatttcgctt tttcccagtg aaacgaagaa
     2941 cgaaccgacc atcatgatat gctcctctgc atgttgcgta ttgaatcaat gacaatttca
     3001 attaagccgc ccgttcgtca tgcgttttcg tgcgcttcga aatgctgata acgctgctgt
     3061 cctccaactg ctttgcatgt ggacacaatt ccatttattt aattctttta tttggatcgg
     3121 ttaaattaaa aagcgccttg ttacgcattt aacgttgttt ccggtgcgtg gtggtttcat
     3181 gcttctggga acggcaaatg ggtttaggat tgggaacccc tcatcatctg ttggaatata
     3241 ctattcaacc tacaaaagta acgttaaaca acactacttt atatttgata tgaatggcca
     3301 caccttttat gccataaaac atattgtaag agaataccac tctttttatt ccttctttcc
     3361 ttcttgtacg ttttttgctg taagtaggtc gtggtgctgg tgttgcagtt gaaataactt
     3421 aaaatataaa tcataaaact caaacataaa cttgactatt tatttattta ttaagaaagg
     3481 aaatataaat tataaattac aacaggttat ggacctgcag ccaagcttgg cgctcgtccg
     3541 ggggcaatga gatatgaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat
     3601 cgaaaagttc gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc
     3661 tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg
     3721 tttctacaaa gatcgttatg tttatcggca ctttgcatcg gccgcgctcc cgattccgga
     3781 agtgcttgac attggggaat tcagcgagag cctgacctat tgcatctccc gccgtgcaca
     3841 gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc agccggtcgc
     3901 ggaggccatg gatgcgatcg ctgcggccga tcttagccag acgagcgggt tcggcccatt
     3961 cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga
     4021 tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca
     4081 ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca
     4141 cgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga
     4201 ctggagcgag gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag
     4261 gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct
     4321 tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca
     4381 gagcttggtt gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat
     4441 cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt
     4501 ctggaccgat ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg
     4561 tccgagggca aaggaataga gtagatgccg accgaacaag agctgatttc gagaacgcct
     4621 cagccagcaa ctcgcgcgag cctagcaagg caaatgcgag agaacggcct tacgcttggt
     4681 ggcacagttc tcgtccacag ttcgctaagc tcgctcggct gggtcgcggg agggccggtc
     4741 gcagtgattc aggcccttct ggattgtgtt ggtccccagg gcacgattgt catgcccacg
     4801 cactcgggtg atctgactga tcccgcagat tggagatcgc cgcccgtgcc tgccgattgg
     4861 gtgcagatct ttgtgaagga accttacttc tgtggtgtga cataattgga caaactacct
     4921 acagagattt aaagctctaa ggtaaatata aaatttttaa gtgtataatg tgttaaacta
     4981 ctgattctaa ttgtttgtgt attttagatt ccaacctatg gaactgatga atgggagcag
     5041 tggtggaatg cctttaatga ggaaaacctg ttttgctcag aagaaatgcc atctagtgat
     5101 gatgaggcta ctgctgactc tcaacattct actcctccaa aaaagaagag aaaggtagaa
     5161 gaccccaagg actttccttc agaattgcta agttttttga gtcatgctgt gtttagtaat
     5221 agaactcttg cttgctttgc tatttacacc acaaaggaaa aagctgcact gctatacaag
     5281 aaaattatgg aaaaatattc tgtaaccttt ataagtaggc ataacagtta taatcataac
     5341 atactgtttt ttcttactcc acacaggcat agagtgtctg ctattaataa ctatgctcaa
     5401 aaattgtgta cctttagctt tttaatttgt aaaggggtta ataaggaata tttgatgtat
     5461 agtgccttga ctagagatca taatcagcca taccacattt gtagaggttt tacttgcttt
     5521 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     5581 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     5641 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     5701 ttatcatgtc tggatcaatt cgttgcagga caggatgtgg tgcccgatgt gactagctct
     5761 ttgctgcagg ccgtcctatc ctctggttcc gataagagac ccagaactcc ggccccccac
     5821 cgcccaccgc cacccccata catatgtggt acgcaagtaa gagtgcctgc gcatgcccca
     5881 tgtgccccac caagagcttt gcatcccata caagtcccca aagtggagaa ccgaaccaat
     5941 tcttcgcggg cagaacaaaa gcttctgcac acgtctccac tcgaatttgg agccggccgg
     6001 cgtgtgcaaa agaggtgaat cgaacgaaag acccgtgtgt aaagccgcgt ttccaaaatg
     6061 tataaaaccg agagcatctg gccaatgtgc atcagttgtg gtcagcagca aaatcaagtg
     6121 aatcatctca gtgcaactaa aggggggatc tcgaggaatt ccattgagaa atgactgagt
     6181 tccggtgctc tcaagtcatt gatctttgtc gacttttatt tggtctctgt aataacgact
     6241 tcaaaaacat taaattctgt tgcgaagcca gtaagctaca aaaagaaaaa acaagagaga
     6301 gtgctatagt cgtatagtat agtttcccga ctatctgata cccattactt atctaggggg
     6361 aatgcgaacc caaaatttta tcagttttct cggatatcga tagatattgg ggaataaatt
     6421 taaataaata aattttgggc gggtttaggg cgtggcaaaa agttttttgg caaatcgcta
     6481 gaaatttaca agacttataa aattatgaaa aaatacaaca aaattttaaa cacgtgggcg
     6541 tgacagtttt gggcggtttt agggcgttag agtaggcgag gacagggtta catcgactag
     6601 gctttgatcc tgatcaagaa tatatatact ttataccgct tccttctaca tgttacctat
     6661 ttttcaacga atctagtata cctttttact gtacgattta tgggtataat aataagctaa
     6721 atcgagacta agtttaaacg ttatatatat tttttttatt ttatgcagga tggcagcgtg
     6781 cagctggccg accactacca gcagaatacc cccatcggcg atggccctgt gctgctgccc
     6841 gataaccact acctgtccac ccagagcgcc ctgtccaagg accccaacga gaagcgcgat
     6901 cacatgatct acttcggctt cgtgaccgcc gccgccatca cccacggcat ggacgagctg
     6961 tacaagtgaa tcgataccgg tttaattaac acgtgggtaa ccgttccgag tcgttaacca
     7021 ccatggccag caagggcgcc gagctgttca ccggcatcgt gcccatcctg atcgagctga
     7081 atggcgatgt gaatggccac aagttcagcg tgagcggcga gggcgagggc gatgccacct
     7141 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcctgtg ccctggccca
     7201 ccctggtgac caccctgagc tacggcgtgc agtgcttctc acgctacccc gatcacatga
     7261 agcagcacga cttcttcaag agcgccatgc ctgagggcta catccaggag cgcaccatct
     7321 tcttcgagga tgacggcaac tacaagtcgc gcgccgaggt gaagttcgag ggcgataccc
     7381 tggtgaatcg catcgagctg accggcaccg atttcaagga ggatggcaac atcctgggca
     7441 ataagatgga gtacaactac aacgcccaca atgtgtacat catgaccgac aaggccaaga
     7501 atggcatcaa ggtgaacttc aagatccgcc acaacatcga ggtaagtatt caaaattcca
     7561 aaattttttc ctaggaatat tcgatttttt aataggcagt ttctatacta ttgtatacta
     7621 ttgtagattc gttgaaaagt atgtaacagg aagaataaag catttccgac catgtaaagt
     7681 atatatattc ttaataagga tcaatagccg agtcgatctc gccatgtccg tctgtcttat
     7741 tattttatta ccgccgagac atcaggaact ataaaagcta gaaggatgag ttttagcata
     7801 cagattctag agacaaggac gcagagcaag tttgttgatc catgctgcca cgctttaact
     7861 ttctcaaatt gcccaaaact gccatgccca catttttgaa ctattttcga aattttttca
     7921 taattgtatt actcgtgtaa atttccatca atttgccaaa aaactttttg tcacgcgtta
     7981 acgccctaaa gccgccaatt tggtcacgcc cacactattg aacaattatc aaattttttc
     8041 tcattttatt ccccaatatc tatcgatatc cccgattatg aaattattaa atttcgcgtt
     8101 cgcattcaca ctagctgagt aacgagtatc tgatagttgg ggaaatcgac ttatttttta
     8161 tatacaatga aaatgaattt aatcatatga atatcgatta tagcttttta tttaatatga
     8221 atatttattt gggcttaagg tgtaacctcc tcgacataag actcacatgg cgcaggcaca
     8281 ttgaagacaa aaatactcat tgtcgggtct cgcaccctcc agcagcacct aaaattatgt
     8341 cttcaattat tgccaacatt ggagacacaa ttagtctgtg gcacctcagg cggccgcaac
     8401 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     8461 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     8521 ctagtggatc cactaggggc cgccgacgcg aggctggatg gccttcccca ttatgattct
     8581 tctcgcttcc ggcggcatcg ggatgcccgc gttgcaggcc atgctgtcca ggcaggtaga
     8641 tgacgaccat cagggacagc ttcaaggatc gctcgcggct cttaccagcc taacttcgat
     8701 cattggaccg ctgatcgtca cggcgattta tgccgcctcg gcgagcacat ggaacgggtt
     8761 ggcatggatt gtaggcgccg ccctatacct tgtctgcctc cccgcgttgc gtcgcggtgc
     8821 atggagccgg gccacctcga cctgaatgga agccggcggc acctcgctaa cggattcacc
     8881 actccaagaa ttggagccaa tcaattcttg cggagaactg tgaatgcgca aaccaaccct
     8941 tggcagaaca tatccatcgc gtccgccatc tccagcagcc gcacgcggcg catctcgggc
     9001 agcgttgggt cctggccacg ggtgcgcatg atcgtgctcc tgtcgttgag gacccggcta
     9061 ggctggcggg gttgccttac tggttagcag aatgaatcac cgatacgcga gcgaacgtga
     9121 agcgactgct gctgcaaaac gtctgcgacc tgagcaacaa catgaatggt cttcggtttc
     9181 cgtgtttcgt aaagtctgga aacgcggaag tcagcgctct tccgcttcct cgctcactga
     9241 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     9301 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     9361 aaaggccagg aaccgta
//
