LOCUS       sequence-124090-        8012 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 8012)
  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 8012)
  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..8012
                     /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    1660..2212
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1662..2235
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1822..1841)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     2051..2068
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     complement(3606..3623)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3612..3836
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3882..4310
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3969..3988)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4179..4200
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4319..4339)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4324..4653
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4504..4639
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4720..5514
                     /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(4774..4793)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5384..5403
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5688..5809
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5725..5744)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5779..5798
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5858..5874)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5858..5874)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5871..5893)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5882..5898
                     /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(5906..5912,5913..5930,5931..5936))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5951..5972
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6089..6106)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6260..6845)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6340..6359)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7016..7807,7808..7876))
                     /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     7639..7658
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7877..7981)
                     /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 ttttgacgga
     1021 gtttaacttc ttgtctccca ggtaggaagt gcagtggcgt aatctcggct cactacaacc
     1081 tccacctcct gggttcaagc gtttctcctg cctcagcttt ccgagtagct gggattacag
     1141 gcgcctgcca ccatgccctg ctgacttttg tatttttagt agagacgggg tttcaccatg
     1201 ttggccaggc tggtcttgaa ctcctgaccg caggcgattg gcctgcctcg gcctcccaaa
     1261 gtgctgagat tacaggcgtg agccaccacc cccggcctca ggagcgttct gatagtgcct
     1321 cgatgtgctg cctcctataa agtgttagca gcacagatca ctttttgtaa aggtacgtac
     1381 taatgacttt ttttttatac ttcaggatgg cagcgtgcag ctggccgacc actaccagca
     1441 gaataccccc atcggcgatg gccctgtgct gctgcccgat aaccactacc tgtccaccca
     1501 gagcgccctg tccaaggacc ccaacgagaa gcgcgatcac atgatctact tcggcttcgt
     1561 gaccgccgcc ggcatcaccc acggcatgga cgagctgtac aagtgaatcg ataccggttt
     1621 aattaacacg tgggtaaccg ttccgagtcg ttaacgccat attatcatcg tgtttttcaa
     1681 aggaaaacca cgtccccgtg gttcgggggg cctagacgtt tttttaacct cgactaaaca
     1741 catgtaaagc atgtgcaccg aggccccaga tcagatccca tacaatgggg taccttctgg
     1801 gcatccttca gccccttgtt gaatacgctt gaggagagcc atttgactct ttccacaact
     1861 atccaactca caacgtggca ctggggttgt gccgcctttg caggtgtatc ttatacacgt
     1921 ggcttttggc cgcagaggca cctgtcgcca ggtggggggt tccgctgcct gcaaagggtc
     1981 gctacagacg ttgtttgtct tcaagaagct tccagaggaa ctgcttcctt cacgacattc
     2041 aacagacctt gcattccttt ggcgagaggg gaaagacccc taggaatgct cgtcaagaag
     2101 acagggccag gtttccgggc cctcacattg ccaaaagacg gcaatatggt ggaaaataac
     2161 atatagacaa acgcacaccg gccttattcc aagcggcttc ggccagtaac gttagggggg
     2221 ggggagggag aggggcgtta accaccatgg ccagcaaggg cgccgagctg ttcaccggca
     2281 tcgtgcccat cctgatcgag ctgaatggcg atgtgaatgg ccacaagttc agcgtgagcg
     2341 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     2401 gcaagctgcc tgtgccctgg cccaccctgg tgaccaccct gagctacggc gtgcagtgct
     2461 tctcacgcta ccccgatcac atgaagcagc acgacttctt caagagcgcc atgcctgagg
     2521 gctacatcca ggagcgcacc atcttcttcg aggatgacgg caactacaag tcgcgcgccg
     2581 aggtgaagtt cgagggcgat accctggtga atcgcatcga gctgaccggc accgatttca
     2641 aggaggatgg caacatcctg ggcaataaga tggagtacaa ctacaacgcc cacaatgtgt
     2701 acatcatgac cgacaaggcc aagaatggca tcaaggtgaa cttcaagatc cgccacaaca
     2761 tcgaggtaag aagcaaggtt tcatttaggg gaagggaaat gattcaggac gagagtcttt
     2821 gtgctgctga gtgcctgtga tgaagaagca tgttagtcct gggcaacgta gcgagacccc
     2881 atctctacaa aaaatagaaa aattagccag gtatagtggc gcacacctgt gattccagct
     2941 acgcaggagg ctgaggtggg aggattgctt gagcccagga ggttgaggct gcagtgagct
     3001 gtaatcatgc cactactcca acctgggcaa cacagcaagg accctgtctc aaaagctact
     3061 tacagaaaag aattaggctc ggcacggtag ctcacacctg taatcccagc actttgggag
     3121 gctgaggcgg gcagatcact tgaggtcagg agtttgagac cagcctggcc aacatggtga
     3181 aaccttgtct ctactaaaaa tatgaaaatt agccaggcat ggtggcacat tcctgtaatc
     3241 ccagctactc gggaggctga ggcaggagaa tcacttgaac ccaggaggtg gaggttgcag
     3301 taagccgaga tcgtaccact gtgctctagc cttggtgaca gagcgagact gtcttaaaaa
     3361 aaaaaaaaaa aaaaaaagaa ttaattaaaa atttaaaaaa aaatgaaaaa aagctgcatg
     3421 cttgtttttt gtttttagtt attctacatt gttgtcatta ttaccaaata ttggggaaaa
     3481 tacaacttac agaccaatct caggagttaa atgttactac gaaggcaaat gaactatgcg
     3541 taatgaacct ggtaggcatt agcggccgct cgagtctaga gggcccgttt aaacccgctg
     3601 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     3661 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     3721 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     3781 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     3841 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     3901 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     3961 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4021 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     4081 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4141 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4201 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     4261 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     4321 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4381 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     4441 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     4501 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     4561 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     4621 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     4681 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     4741 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     4801 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     4861 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     4921 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     4981 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     5041 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     5101 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     5161 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     5221 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     5281 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     5341 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     5401 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     5461 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     5521 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     5581 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     5641 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
     5701 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     5761 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
     5821 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     5881 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     5941 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     6001 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     6061 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     6121 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     6181 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     6241 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6301 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6361 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6421 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     6481 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     6541 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     6601 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     6661 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     6721 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     6781 ccaccgctgg tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     6841 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     6901 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     6961 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     7021 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     7081 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     7141 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     7201 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     7261 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     7321 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     7381 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     7441 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     7501 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     7561 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     7621 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     7681 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     7741 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     7801 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     7861 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     7921 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     7981 gcacatttcc ccgaaaagtg ccacctgacg tc
//
