LOCUS       sequence-135398-        6798 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of circular RNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6798)
  AUTHORS   Zhang XO, Wang HB, Zhang Y, Lu X, Chen LL, Yang L
  TITLE     Complementary sequence-mediated exon circularization.
  JOURNAL   Cell. 2014 Sep 25;159(1):134-47. doi: 10.1016/j.cell.2014.09.001. 
            Epub 2014 Sep 18.
  PUBMED    25242744
REFERENCE   2  (bases 1 to 6798)
  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"; doi: 
            "10.1016/j.cell.2014.09.001"; journalName: "Cell"; date: 
            "2014-09-25- 25"; volume: "159"; issue: "1"; pages: "134-47"
COMMENT     Sequence Label: pZW1-FCS-circRNA
FEATURES             Location/Qualifiers
     source          1..6798
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3044..3063)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3391..3412
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3587..3707
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3624..3643)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3677..3696
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3714..4169)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3851..3872)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4063..4082
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4196..4300
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4302..4659
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4302..4493
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4325..4345)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4510..4645
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4694..5488
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4694..5488
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4748..4767)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5358..5377
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5676..5695)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5720..5767
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6096..6684
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6585..6604
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggca
      901 cgtagcaggc cagggcctct ctcagccacc tgagcagaaa gctttccaag atagggcagg
      961 ctgggttagg ccatctgagt ctgtctcgtt cattgggatc cagacttgac tgtcttgtta
     1021 aaggctgttg ctgcccaggt gtgcagggag ctgttggtct ctggcattca gggtgggggt
     1081 ggtataaacc cggggcagct tgcatatggc agggaagagg gatccgtgga ggaacagtgc
     1141 agaaggcttt atgttcagaa tctctcttgc ttttcttcta gactgagttc cttgagattg
     1201 gtgaatgctg tgtattattc atccctgata acctggtgtt tggcccaggg ccttgtccag
     1261 aggagtgttt gataagtgtt tcaagtgaat tagcaccacg atgtcatctc ttttcagttt
     1321 acaaaggacg gacaccccct gacccggtct cagaaagcct gaaagcagaa ttagtcatta
     1381 gaagggtggt tggcttggtc ggcatagact ttgagcagaa agaggttgaa aatgttgagc
     1441 ctgatttctc ttaggcccct ctgcagtgtc tgttgtggag gccagatacg taactgcttc
     1501 cgcttttttt ggtctcattc aaggtgagca aatccccttc atgtttctca ccagacaatg
     1561 cagctgatga ggttccagct ttgcaaatgt agtcatccat gaggactgtc ttcctgagat
     1621 ttcatcaggc tcgagggact tgcaaaggac tttaggtcca ttgtcctttt attcttagat
     1681 acctctttca ctgagacctt ttccttacct cacctctcta ggtggaacgg cacatgtgtg
     1741 atggggacat tgttatcttc aaccggcagc caactctgca caaaatgtcc atgatggggc
     1801 atcgggtccg cattctccca tggtctacct ttcgcttgaa tcttaggtca gtccctggct
     1861 gagggaagca ggctggaatt ggtgggaggc gggcaggctg ggtggctcct caaggtttcg
     1921 ctgcagacat cttcccaacc ctgacttttc tctttaactg tagtgtgaca actccgtaca
     1981 atgcagactt tgacggggat gagatgaact tgcacctgcc acagtctctg gagacgcgag
     2041 cagagatcca ggagctggcc atggttcctc gcatgattgt caccccccag agcaatcggc
     2101 ctgtcatggg tattgtgcag gacacactca cagcagtgcg caaattcacc aagagagacg
     2161 tcttcctgga gcgggtgtgt ggtccaaatg gaaacctggc ttaagtgggc agtggggctc
     2221 tggggtgcaa ggtggaggct agagaggaag agctgtgttt tttttcctga cttacccagc
     2281 agtggtctgt gagattgtct tttctggtgg gcgaacaaaa agggggttag gaaaactcag
     2341 gccaaaaaag tgtaaggcgt taattcccca tttaattcct taaaatttca tgtaatacca
     2401 ggtattgcct gtaaaggaaa gataaaggga aaaataagta agaccttgtt aaaattttat
     2461 ttttctattt taaccttcac ttatttccta attattaaaa gaaatttatg cttattgtta
     2521 agaacaaaaa aatttcagta ttacaatgaa tttttaatta aaagtttttg gcctgatgaa
     2581 atctcaggaa gacagtcctc atggatgact acatttgcaa agctggaacc tcatcagctg
     2641 cattgtctgg tgagaaacat gaaggggatt tgctcacctt gaatgagacc aaaaaaagcg
     2701 gaagcagtta cgtatctggc ctccacaaca gacactgcag aggggcctaa gagaaatcag
     2761 gctcaacatt ttcaacctct ttctgctcaa agtctatgcc gaccaagcca accacccttc
     2821 taatgactaa ttctgctttc aggctttctg agaccgggtc agggggtgtc cttaaaggtt
     2881 ggaaaaaact tttcctgtca tctttgcctc caaaatctgg ctttctccct tgggcaggga
     2941 aacctcccca acatttctct atcatccctg agatgtgggg cctgcactct gacttctgtc
     3001 tgccttactc tttgtcttac aggagcgcac catcttcttc aaggacgacg gcaactacaa
     3061 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     3121 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     3181 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     3241 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     3301 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
     3361 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     3421 cgggatcact ctcggcatgg acgagctgta caagtaagga tccaccggat ctagataact
     3481 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     3541 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     3601 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     3661 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt
     3721 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     3781 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     3841 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     3901 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     3961 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     4021 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     4081 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     4141 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     4201 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4261 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     4321 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     4381 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     4441 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4501 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4561 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     4621 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
     4681 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     4741 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     4801 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     4861 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     4921 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     4981 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     5041 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     5101 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     5161 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     5221 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     5281 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     5341 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     5401 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     5461 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     5521 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     5581 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     5641 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     5701 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     5761 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     5821 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     5881 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     5941 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     6001 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     6061 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6121 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6181 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6241 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6301 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6361 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6421 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6481 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6541 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6601 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6661 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6721 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     6781 cgtattaccg ccatgcat
//
