LOCUS       sequence-127150-        8150 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the human ciRS7/CDR1as circular RNA in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8150)
  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 8150)
  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-ciRS7 Exon
FEATURES             Location/Qualifiers
     source          1..8150
                     /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(3744..3761)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3750..3974
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4020..4448
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4107..4126)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4317..4338
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4457..4477)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4462..4791
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4642..4777
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4858..5652
                     /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(4912..4931)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5522..5541
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5826..5947
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5863..5882)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5917..5936
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5996..6012)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5996..6012)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6009..6031)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6020..6036
                     /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(6044..6050,6051..6068,6069..6074))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6089..6110
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6227..6244)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6398..6983)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6478..6497)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7154..7945,7946..8014))
                     /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     7777..7796
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8015..8119)
                     /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 tcaggatatc ggtttccgat ggcacctgtg tcaaggtctt
     1441 ccaacaactc cgggtcttcc agcgacttca agtcttccaa taatctcaag gtcttccaga
     1501 taatcctgag cttccagaaa atccacatct tccagacaat ccatgtcttc cggacaatcc
     1561 atgtcttcca agaagctcca agtcttccag taaatcaagt cttccagcaa atccagtctt
     1621 ccagcaatta ctggtcttcc accaaatcca gatcttccag gaaaatccac gtcttccagg
     1681 aaatccatgt cttccaataa tttcaaggtc ttccatcaaa tacagatctt ccagctaatc
     1741 catgtcttcc agaaaaatct gtgtcttcca ccaaatccaa gtcttccagt aaatctagtt
     1801 cttccagaaa aatctagatc ttccagtcaa tcagtgtctt ccagaaagaa atccaggtct
     1861 tccagtcaat cagtgtcttc cagaaagaaa tccaggtctt ccagtcagtc agtgtcttcc
     1921 agaaaaatct acgtcttcca ccaaatccag gtcttccagt caatccacat cttccggaaa
     1981 aaatccaggt cttccagcca atatatgtct tcctgaagat ccacgtcttc cagaaaatcc
     2041 atgtcttcca gaaaatccat gtcttccagt aacctcccag tcttccagaa aatccacgtc
     2101 ttcccaacaa tccaagtctt ccggataatt tgggtcttcc tgaaaatcta cgtcttccaa
     2161 aaaagccatg tcttccagaa aatccacatc ttccaatggc ctccaggtct tccagactat
     2221 ccatgtcttc cagaaaatcc ttgtcttccc ttaaatctat agcttccaaa aaatccgggt
     2281 cttccaggaa atccgtgtct tccagcaagt ccacgtcttc caacaaagcc atgtcttcca
     2341 gactatccat gtcttccaga aaatccttgt cttccctcaa atccatagct tccgaaaaat
     2401 ccaggtcttc caggaaatcc gtgtcttcca gcaaatccac gtcttccaac aaagccatgt
     2461 cttccatcaa attaatgtct tccagcctac ttgtgtcttc caacaaaggt acgtcttcca
     2521 acaaaggtac gtcttccaac aaaggtatgt cttccaacaa aggtacgtct tccagaaaat
     2581 ccacgtcttc caaccaagcc atgtcttcca gaaaatccac gtcttccaga aaatatatgt
     2641 cttccaacta agctacgtct tccaacaaat ccatgtcttc ctatatctcc aggtcttcca
     2701 gcatctccag ggcttccagc atctgctcgt cttccaacat ctccacgtct tccagcatct
     2761 ctgtgtcttc cagcatcttc atgtcttcca acaactaccc agtcttccat caactggctc
     2821 aatatccatg tcttccaacg tctccagtgt gctgatcttc tgacattcag gtcttccagt
     2881 gtctgcaata tccagccgcg gaggtaagaa gcaaggtttc atttagggga agggaaatga
     2941 ttcaggacga gagtctttgt gctgctgagt gcctgtgatg aagaagcatg ttagtcctgg
     3001 gcaacgtagc gagaccccat ctctacaaaa aatagaaaaa ttagccaggt atagtggcgc
     3061 acacctgtga ttccagctac gcaggaggct gaggtgggag gattgcttga gcccaggagg
     3121 ttgaggctgc agtgagctgt aatcatgcca ctactccaac ctgggcaaca cagcaaggac
     3181 cctgtctcaa aagctactta cagaaaagaa ttaggctcgg cacggtagct cacacctgta
     3241 atcccagcac tttgggaggc tgaggcgggc agatcacttg aggtcaggag tttgagacca
     3301 gcctggccaa catggtgaaa ccttgtctct actaaaaata tgaaaattag ccaggcatgg
     3361 tggcacattc ctgtaatccc agctactcgg gaggctgagg caggagaatc acttgaaccc
     3421 aggaggtgga ggttgcagta agccgagatc gtaccactgt gctctagcct tggtgacaga
     3481 gcgagactgt cttaaaaaaa aaaaaaaaaa aaaaagaatt aattaaaaat ttaaaaaaaa
     3541 atgaaaaaaa gctgcatgct tgttttttgt ttttagttat tctacattgt tgtcattatt
     3601 accaaatatt ggggaaaata caacttacag accaatctca ggagttaaat gttactacga
     3661 aggcaaatga actatgcgta atgaacctgg taggcattag cggccgctcg agtctagagg
     3721 gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     3781 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     3841 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     3901 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     3961 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     4021 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4081 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4141 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     4201 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     4261 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4321 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     4381 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4441 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     4501 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     4561 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4621 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4681 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     4741 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     4801 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     4861 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     4921 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     4981 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     5041 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     5101 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     5161 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     5221 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     5281 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     5341 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     5401 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     5461 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     5521 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     5581 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     5641 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     5701 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     5761 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     5821 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     5881 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     5941 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     6001 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     6061 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     6121 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     6181 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     6241 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     6301 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     6361 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     6421 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6481 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6541 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6601 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6661 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6721 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6781 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     6841 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6901 cttgatccgg caaacaaacc accgctggta gcggtttttt tgtttgcaag cagcagatta
     6961 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     7021 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     7081 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     7141 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     7201 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     7261 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     7321 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     7381 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     7441 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     7501 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     7561 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     7621 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     7681 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     7741 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     7801 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     7861 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     7921 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     7981 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     8041 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     8101 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
