LOCUS       sequence-180700-        7897 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a non-polyadenylated, translatable EGFP mRNA in 
            Drosophila.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7897)
  AUTHORS   Liang D, Tatomer DC, Luo Z, Wu H, Yang L, Chen LL, Cherry S, Wilusz 
            JE
  TITLE     The Output of Protein-Coding Genes Shifts to Circular RNAs When the 
            Pre-mRNA Processing Machinery Is Limiting.
  JOURNAL   Mol Cell. 2017 Nov 9. pii: S1097-2765(17)30835-3. doi: 
            10.1016/j.molcel.2017.10.034.
  PUBMED    29174924
REFERENCE   2  (bases 1 to 7897)
  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: "Mol Cell. 
            2017 Nov 9. pii: S1097-2765(17)30835-3. doi: 
            10.1016/j.molcel.2017.10.034."
COMMENT     Sequence Label: Hy_pMT eGFP-RVFV NSs HhRz
FEATURES             Location/Qualifiers
     source          1..7897
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        complement(139..190)
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             complement(join(273..986,987..989,990..992))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(276..986,987..989,990..992))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(318..339)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     667..686
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     926..947
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1010..1026)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1011..1027)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(1035..1461)
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     polyA_signal    1462..1596
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1486..1505)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1540..1559
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2021..2041)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          2171..2236
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(2599..3624)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(2599..3624)
                     /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"
     promoter        complement(3670..3949)
                     /label=copia promoter
                     /note="strong promoter from the Drosophila transposable 
                     element copia (Sinclair et al., 1986)"
     primer_bind     complement(5345..5363)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5431..5535
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5536..5604,5605..6396)
                     /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     complement(5754..5773)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6567..7155
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7056..7075
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7309..7326
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gtccctgatg gtcgtcatct acctgcctgg acagcatggc ctgcaacgcg ggcatcccga
       61 tgccgccgga agcgagaaga atcataatgg ggaaggccat ccagcctcgc gtcggcggcc
      121 cctagtggat ccactagtcc tgtttcgtcc tcacggactc atcagaccgg aaaacacatc
      181 cggtgacagg ccagggggtt ggggggaagg ggagttgggg ttacggtcgg gattaggggg
      241 tggggggtgg ggcagcctta acctgcggcc gcttacttgt acagctcgtc catgccgaga
      301 gtgatcccgg cggcggtcac gaactccagc aggaccatgt gatcgcgctt ctcgttgggg
      361 tctttgctca gggcggactg ggtgctcagg tagtggttgt cgggcagcag cacggggccg
      421 tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct gcacgctgcc gtcctcgatg
      481 ttgtggcgga tcttgaagtt caccttgatg ccgttcttct gcttgtcggc catgatatag
      541 acgttgtggc tgttgtagtt gtactccagc ttgtgcccca ggatgttgcc gtcctccttg
      601 aagtcgatgc ccttcagctc gatgcggttc accagggtgt cgccctcgaa cttcacctcg
      661 gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg tgcgctcctg gacgtagcct
      721 tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta gcggctgaag
      781 cactgcacgc cgtaggtcag ggtggtcacg agggtgggcc agggcacggg cagcttgccg
      841 gtggtgcaga tgaacttcag ggtcagcttg ccgtaggtgg catcgccctc gccctcgccg
      901 gacacgctga acttgtggcc gtttacgtcg ccgtccagct cgaccaggat gggcaccacc
      961 ccggtgaaca gctcctcgcc cttgctcacc atggtgatat caagcttatc gataccgtcg
     1021 acctcgagat ccccccttta gttgcactga gatgattcac ttgattttgc tgctgaccac
     1081 aactgatgca cattggccag atgctctcgg ttttatacat tttggaaacg cggctttaca
     1141 cacgggtctt tcgttcgatt cacctctttt gcacacgccg gccggctcca aattcgagtg
     1201 gagacgtgtg cagaagcttt tgttctgccc gcgaagaatt ggttcggttc tccactttgg
     1261 ggacttgtat gggatgcaaa gctcttggtg gggcacatgg ggcatgcgca ggcactctta
     1321 cttgcgtacc acatatgtat gggggtggcg gtgggcggtg gggggccgga gttctgggtc
     1381 tcttatcgga accagaggat aggacggcct gcagcaaaga gctagtcaca tcgggcacca
     1441 catcctgtcc tgcaacgaat tgatccagac atgataagat acattgatga gtttggacaa
     1501 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     1561 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     1621 atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa
     1681 tgtggtatgg ctgattatga tctctagtca aggcactata catcaaatat tccttattaa
     1741 cccctttaca aattaaaaag ctaaaggtac acaatttttg agcatagtta ttaatagcag
     1801 acactctatg cctgtgtgga gtaagaaaaa acagtatgtt atgattataa ctgttatgcc
     1861 tacttataaa ggttacagaa tatttttcca taattttctt gtatagcagt gcagcttttt
     1921 cctttgtggt gtaaatagca aagcaagcaa gagttctatt actaaacaca gcatgactca
     1981 aaaaacttag caattctgaa ggaaagtcct tggggtcttc tacctttctc ttcttttttg
     2041 gaggagtaga atgttgagag tcagcagtag cctcatcatc actagatggc atttcttctg
     2101 agcaaaacag gttttcctca ttaaaggcat tccaccactg ctcccattca tcagttccat
     2161 aggttggaat ctaaaataca caaacaatta gaatcagtag tttaacacat tatacactta
     2221 aaaattttat atttacctta gagctttaaa tctctgtagg tagtttgtcc aattatgtca
     2281 caccacagaa gtaaggttcc ttcacaaaga tctgcaccca atcggcaggc acgggcggcg
     2341 atctccaatc tgcgggatca gtcagatcac ccgagtgcgt gggcatgaca atcgtgccct
     2401 ggggaccaac acaatccaga agggcctgaa tcactgcgac cggccctccc gcgacccagc
     2461 cgagcgagct tagcgaactg tggacgagaa ctgtgccacc aagcgtaagg ccgttctctc
     2521 gcatttgcct tgctaggctc gcgcgagttg ctggctgagg cgttctcgaa atcagctctt
     2581 gttcggtcgg catctactct attcctttgc cctcggacga gtgctggggc gtcggtttcc
     2641 actatcggcg agtacttcta cacagccatc ggtccagacg gccgcgcttc tgcgggcgat
     2701 ttgtgtacgc ccgacagtcc cggctccgga tcggacgatt gcgtcgcatc gaccctgcgc
     2761 ccaagctgca tcatcgaaat tgccgtcaac caagctctga tagagttggt caagaccaat
     2821 gcggagcata tacgcccgga gccgcggcga tcctgcaagc tccggatgcc tccgctcgaa
     2881 gtagcgcgtc tgctgctcca tacaagccaa ccacggcctc cagaagaaga tgttggcgac
     2941 ctcgtattgg gaatccccga acatcgcctc gctccagtca atgaccgctg ttatgcggcc
     3001 attgtccgtc aggacattgt tggagccgaa atccgcgtgc acgaggtgcc ggacttcggg
     3061 gcagtcctcg gcccaaagca tcagctcatc gagagcctgc gcgacggacg cactgacggt
     3121 gtcgtccatc acagtttgcc agtgatacac atggggatca gcaatcgcgc atatgaaatc
     3181 acgccatgta gtgtattgac cgattccttg cggtccgaat gggccgaacc cgctcgtctg
     3241 gctaagatcg gccgcagcga tcgcatccat ggcctccgcg accggctgca gaacagcggg
     3301 cagttcggtt tcaggcaggt cttgcaacgt gacaccctgt gcacggcggg agatgcaata
     3361 ggtcaggctc tcgctgaatt ccccaatgtc aagcacttcc ggaatcggga gcgcggccga
     3421 tgcaaagtgc cgataaacat aacgatcttt gtagaaacca tcggcgcagc tatttacccg
     3481 caggacatat ccacgccctc ctacatcgaa gctgaaagca cgagattctt cgccctccga
     3541 gagctgcatc aggtcggaga cgctgtcgaa cttttcgatc agaaacttct cgacagacgt
     3601 cgcggtgagt tcaggctttt tcatatctca ttgcccccgg acgagcgcca agcttggctg
     3661 caggtccata acctgttgta atttataatt tatatttcct ttcttaataa ataaataaat
     3721 agtcaagttt atgtttgagt tttatgattt atattttaag ttatttcaac tgcaacacca
     3781 gcaccacgac ctacttacag caaaaaacgt acaagaagga aagaaggaat aaaaagagtg
     3841 gtattctctt acaatatgtt ttatggcata aaaggtgtgg ccattcatat caaatataaa
     3901 gtagtgttgt ttaacgttac ttttgtaggt tgaatagtat attccaacag atgatgaggg
     3961 gttcccaatc ctaaacccat ttgccgttcc cagaagcatg aaaccaccac gcaccggaaa
     4021 caacgttaaa tgcgtaacaa ggcgcttttt aatttaaccg atccaaataa aagaattaaa
     4081 taaatggaat tgtgtccaca tgcaaagcag ttggaggaca gcagcgttat cagcatttcg
     4141 aagcgcacga aaacgcatga cgaacgggcg gcttaattga aattgtcatt gattcaatac
     4201 gcaacatgca gaggagcata tcatgatggt cggttcgttc ttcgtttcac tgggaaaaag
     4261 cgaaattgaa tccattttga ttgcgtttgt gtcgcaagag caacacattg cctgcgccac
     4321 atcgcgttcg cttatcaacg ggcgaagaaa ggaaaatgta catttaattg gtcgtaaaaa
     4381 agtgcattgt aaatgttaaa gtcgctgata gcttcgggcc gcttgttcgc aatgataacg
     4441 acagtgtaac aaattccaga ggggagcccc ttccaaaata cgagattcca aaaaaaaaaa
     4501 aacaaaaaaa agaagaagcg tttcgaaact ctgaagaaaa cccagagaag ataagagcgt
     4561 acttgacagc atatagcaaa actattgtac atttccaaat ttcaattgga atcgaaacgg
     4621 gaacagtcaa tcgcttgggt tatcggaata atcgcctcaa ttgcgtgaac ctatcgacac
     4681 aaagtgcatt ggtattgagt caaagtaagg tttgcttata ttaactttca tagattgcat
     4741 atttttgccg gcttattcat gggaataaga aatgtttgag ctgaactcga tagctcaaca
     4801 ataacacgac ttggtgtcaa ttaaatgata atacgtgact agaccatgga aattcaatgt
     4861 attgctattg gatacaatac atatacaagt agttgttgaa aagaaagcag cgaaaactgg
     4921 tcaggtaatg agtcatcatc atcggagtca tcttgactga ttagcacaaa agccagttct
     4981 catctgtatg caaatggctt aaacgcttct acgtacataa gtagttgatg ttgttaaaca
     5041 ggaacttaag atgccgccaa caacagtttc cacagcaatt aaacccaaat ttttaattgc
     5101 gcataacagg cgtctttcat gttcctgtcc gcatttgaca ttgatctcct tctgaatcct
     5161 gattcgataa tatgtaatct tggggccatc aatcacgcag tcagcagcga tttgatatat
     5221 gtatatataa caccatatta ttcaaagccg tttgccttgc tcggttgatg aggaaattgg
     5281 tgaaaaactc gaacttttga taaaaaaaaa attcgtacaa atgtgaattc ttgaagacga
     5341 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     5401 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     5461 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     5521 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     5581 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     5641 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     5701 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     5761 ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat
     5821 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     5881 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     5941 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     6001 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     6061 cgtgacacca cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa
     6121 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     6181 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     6241 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     6301 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     6361 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     6421 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     6481 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     6541 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     6601 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     6661 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     6721 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     6781 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     6841 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     6901 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     6961 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     7021 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     7081 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     7141 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     7201 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     7261 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     7321 agcgaggaag cggaagagcg ctgacttccg cgtttccaga ctttacgaaa cacggaaacc
     7381 gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt
     7441 tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg
     7501 ggtcctcaac gacaggagca cgatcatgcg cacccgtggc caggacccaa cgctgcccga
     7561 gatgcgccgc gtgcggctgc tggagatggc ggacgcgatg gatatgttct gccaagggtt
     7621 ggtttgcgca ttcacagttc tccgcaagaa ttgattggct ccaattcttg gagtggtgaa
     7681 tccgttagcg aggtgccgcc ggcttccatt caggtcgagg tggcccggct ccatgcaccg
     7741 cgacgcaacg cggggaggca gacaaggtat agggcggcgc ctacaatcca tgccaacccg
     7801 ttccatgtgc tcgccgaggc ggcataaatc gccgtgacga tcagcggtcc aatgatcgaa
     7861 gttaggctgg taagagccgc gagcgatcct tgaagct
//
