LOCUS       sequence-129567-        8011 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a GFP circular RNA containing the antisense EMCV IRES in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8011)
  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 8011)
  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: pcDNA3.1(+) ZKSCAN1 MCS-WT Split GFP + Antisen...
FEATURES             Location/Qualifiers
     source          1..8011
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    1659..2211
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1661..2234
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1821..1840)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     2050..2067
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     complement(3605..3622)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3611..3835
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3881..4309
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3968..3987)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4178..4199
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4318..4338)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4323..4652
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4503..4638
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4719..5513
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4773..4792)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5383..5402
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5687..5808
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5724..5743)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5778..5797
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5857..5873)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5857..5873)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5870..5892)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5881..5897
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5905..5911,5912..5929,5930..5935))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5950..5971
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6088..6105)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6259..6844)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6339..6358)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7015..7806,7807..7875))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7638..7657
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7876..7980)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcagt
      961 gacagtggag attgtacagt tttttcctcg atttgtcagg attttttttt tttgacggag
     1021 tttaacttct tgtctcccag gtaggaagtg cagtggcgta atctcggctc actacaacct
     1081 ccacctcctg ggttcaagcg tttctcctgc ctcagctttc cgagtagctg ggattacagg
     1141 cgcctgccac catgccctgc tgacttttgt atttttagta gagacggggt ttcaccatgt
     1201 tggccaggct ggtcttgaac tcctgaccgc aggcgattgg cctgcctcgg cctcccaaag
     1261 tgctgagatt acaggcgtga gccaccaccc ccggcctcag gagcgttctg atagtgcctc
     1321 gatgtgctgc ctcctataaa gtgttagcag cacagatcac tttttgtaaa ggtacgtact
     1381 aatgactttt tttttatact tcaggatggc agcgtgcagc tggccgacca ctaccagcag
     1441 aataccccca tcggcgatgg ccctgtgctg ctgcccgata accactacct gtccacccag
     1501 agcgccctgt ccaaggaccc caacgagaag cgcgatcaca tgatctactt cggcttcgtg
     1561 accgccgccg ccatcaccca cggcatggac gagctgtaca agtgaatcga taccggttta
     1621 attaacacgt gggtaaccgt tccgagtcgt taacgccata ttatcatcgt gtttttcaaa
     1681 ggaaaaccac gtccccgtgg ttcggggggc ctagacgttt ttttaacctc gactaaacac
     1741 atgtaaagca tgtgcaccga ggccccagat cagatcccat acaatggggt accttctggg
     1801 catccttcag ccccttgttg aatacgcttg aggagagcca tttgactctt tccacaacta
     1861 tccaactcac aacgtggcac tggggttgtg ccgcctttgc aggtgtatct tatacacgtg
     1921 gcttttggcc gcagaggcac ctgtcgccag gtggggggtt ccgctgcctg caaagggtcg
     1981 ctacagacgt tgtttgtctt caagaagctt ccagaggaac tgcttccttc acgacattca
     2041 acagaccttg cattcctttg gcgagagggg aaagacccct aggaatgctc gtcaagaaga
     2101 cagggccagg tttccgggcc ctcacattgc caaaagacgg caatatggtg gaaaataaca
     2161 tatagacaaa cgcacaccgg ccttattcca agcggcttcg gccagtaacg ttaggggggg
     2221 gggagggaga ggggcgttaa ccaccatggc cagcaagggc gccgagctgt tcaccggcat
     2281 cgtgcccatc ctgatcgagc tgaatggcga tgtgaatggc cacaagttca gcgtgagcgg
     2341 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
     2401 caagctgcct gtgccctggc ccaccctggt gaccaccctg agctacggcg tgcagtgctt
     2461 ctcacgctac cccgatcaca tgaagcagca cgacttcttc aagagcgcca tgcctgaggg
     2521 ctacatccag gagcgcacca tcttcttcga ggatgacggc aactacaagt cgcgcgccga
     2581 ggtgaagttc gagggcgata ccctggtgaa tcgcatcgag ctgaccggca ccgatttcaa
     2641 ggaggatggc aacatcctgg gcaataagat ggagtacaac tacaacgccc acaatgtgta
     2701 catcatgacc gacaaggcca agaatggcat caaggtgaac ttcaagatcc gccacaacat
     2761 cgaggtaaga agcaaggttt catttagggg aagggaaatg attcaggacg agagtctttg
     2821 tgctgctgag tgcctgtgat gaagaagcat gttagtcctg ggcaacgtag cgagacccca
     2881 tctctacaaa aaatagaaaa attagccagg tatagtggcg cacacctgtg attccagcta
     2941 cgcaggaggc tgaggtggga ggattgcttg agcccaggag gttgaggctg cagtgagctg
     3001 taatcatgcc actactccaa cctgggcaac acagcaagga ccctgtctca aaagctactt
     3061 acagaaaaga attaggctcg gcacggtagc tcacacctgt aatcccagca ctttgggagg
     3121 ctgaggcggg cagatcactt gaggtcagga gtttgagacc agcctggcca acatggtgaa
     3181 accttgtctc tactaaaaat atgaaaatta gccaggcatg gtggcacatt cctgtaatcc
     3241 cagctactcg ggaggctgag gcaggagaat cacttgaacc caggaggtgg aggttgcagt
     3301 aagccgagat cgtaccactg tgctctagcc ttggtgacag agcgagactg tcttaaaaaa
     3361 aaaaaaaaaa aaaaaagaat taattaaaaa tttaaaaaaa aatgaaaaaa agctgcatgc
     3421 ttgttttttg tttttagtta ttctacattg ttgtcattat taccaaatat tggggaaaat
     3481 acaacttaca gaccaatctc aggagttaaa tgttactacg aaggcaaatg aactatgcgt
     3541 aatgaacctg gtaggcatta gcggccgctc gagtctagag ggcccgttta aacccgctga
     3601 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     3661 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     3721 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     3781 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     3841 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     3901 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     3961 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     4021 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     4081 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     4141 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     4201 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     4261 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     4321 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4381 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     4441 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     4501 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     4561 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     4621 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     4681 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     4741 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     4801 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     4861 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     4921 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4981 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     5041 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     5101 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     5161 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     5221 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     5281 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     5341 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     5401 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     5461 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     5521 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     5581 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     5641 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     5701 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     5761 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     5821 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     5881 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     5941 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     6001 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     6061 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     6121 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     6181 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     6241 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     6301 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     6361 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     6421 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     6481 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6541 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6601 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6661 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     6721 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6781 caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6841 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6901 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6961 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     7021 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     7081 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     7141 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     7201 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     7261 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     7321 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     7381 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     7441 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     7501 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     7561 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     7621 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     7681 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     7741 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     7801 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7861 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7921 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     7981 cacatttccc cgaaaagtgc cacctgacgt c
//
