LOCUS       sequence-140445-        6926 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expression plasmid for expressing circular RNAs of a desired 
            sequence in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6926)
  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 6926)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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(+) Laccase2 MCS Exon Vector
FEATURES             Location/Qualifiers
     source          1..6926
                     /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"
     primer_bind     complement(2520..2537)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2526..2750
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2796..3224
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2883..2902)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3093..3114
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3233..3253)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3238..3567
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3418..3553
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3480..3499
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3634..4428
                     /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(3688..3707)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4298..4317
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4602..4723
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4639..4658)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4693..4712
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4772..4788)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4772..4788)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4785..4807)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4796..4812
                     /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(4820..4826,4827..4844,4845..4850))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4865..4886
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5003..5020)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5174..5759)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5254..5273)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5930..6721,6722..6790))
                     /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     6553..6572
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6791..6895)
                     /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 ggaattccat
      961 tgagaaatga ctgagttccg gtgctctcaa gtcattgatc tttgtcgact tttatttggt
     1021 ctctgtaata acgacttcaa aaacattaaa ttctgttgcg aagccagtaa gctacaaaaa
     1081 gaaaaaacaa gagagaatgc tatagtcgta tagtatagtt tcccgactat ctgataccca
     1141 ttacttatct agggggaatg cgaacccaaa attttatcag ttttctcgga tatcgataga
     1201 tattggggaa taaatttaaa taaataaatt ttgggcgggt ttagggcgtg gcaaaaagtt
     1261 ttttggcaaa tcgctagaaa tttacaagac ttataaaatt atgaaaaaat acaacaaaat
     1321 tttaaacacg tgggcgtgac agttttgggc ggttttaggg cgttagagta ggcgaggaca
     1381 gggttacatc gactaggctt tgatcctgat caagaatata tatactttat accgcttcct
     1441 tctacatgtt acctattttt caacgaatct agtatacctt tttactgtac gatttatggg
     1501 tataataata agctaaatcg agactaagtt ttattgttat atatattttt tttattttat
     1561 gcagaaatta attaaaccgg tcctgcaggt gatcaggcgc gccggttacc ggccggcccc
     1621 gcggagcgta agtattcaaa attccaaaat tttttactag aaatattcga ttttttaata
     1681 ggcagtttct atactattgt atactattgt agattcgttg aaaagtatgt aacaggaaga
     1741 ataaagcatt tccgaccatg taaagtatat atattcttaa taaggatcaa tagccgagtc
     1801 gatctcgcca tgtccgtctg tcttattgtt ttattaccgc cgagacatca ggaactataa
     1861 aagctagaag gatgagtttt agcatacaga ttctagagac aaggacgcag agcaagtttg
     1921 ttgatccatg ctgccacgct ttaactttct caaattgccc aaaactgcca tgcccacatt
     1981 tttgaactat tttcgaaatt ttttcataat tgtattactc gtgtaaattt ccatcaattt
     2041 gccaaaaaac tttttgtcac gcgttaacgc cctaaagccg ccaatttggt cacgcccaca
     2101 ctattgagca attatcaaat tttttctcat tttattcccc aatatctatc gatatccccg
     2161 attatgaaat tattaaattt cgcgttcgca ttcacactag ctgagtaacg agtatctgat
     2221 agttggggaa atcgacttat tttttatata caatgaaaat gaatttaatc atatgaatat
     2281 cgattatagc tttttattta atatgaatat ttatttgggc ttaaggtgta acctcctcga
     2341 cataagactc acatggcgca ggcacattga agacaaaaat actcattgtc gggtctcgca
     2401 ccctccagca gcacctaaaa ttatgtcttc aattattgcc aacattggag acacaattag
     2461 tctgtggcac ctcaggcggc cgctcgagtc tagagggccc gtttaaaccc gctgatcagc
     2521 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     2581 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     2641 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     2701 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     2761 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     2821 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     2881 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     2941 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     3001 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     3061 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     3121 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     3181 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     3241 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3301 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3361 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3421 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3481 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     3541 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     3601 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     3661 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     3721 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     3781 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     3841 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     3901 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     3961 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     4021 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     4081 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     4141 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     4201 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     4261 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     4321 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     4381 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     4441 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     4501 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     4561 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     4621 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     4681 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     4741 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     4801 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     4861 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     4921 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     4981 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     5041 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     5101 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     5161 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     5221 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     5281 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5341 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     5401 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5461 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5521 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5581 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     5641 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5701 ctggtagcgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5761 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5821 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     5881 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     5941 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     6001 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     6061 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     6121 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     6181 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     6241 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     6301 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6361 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6421 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     6481 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     6541 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     6601 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     6661 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     6721 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6781 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6841 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     6901 ttccccgaaa agtgccacct gacgtc
//
