LOCUS       sequence-127079-        7183 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a miniature version of the ZKSCAN1 circular RNA in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7183)
  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 7183)
  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(+) Laccase2 MCS-ZKSCAN1 548-847
FEATURES             Location/Qualifiers
     source          1..7183
                     /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(2777..2794)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2783..3007
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3053..3481
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3140..3159)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3350..3371
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3490..3510)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3495..3824
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3675..3810
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3737..3756
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3891..4685
                     /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(3945..3964)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4555..4574
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4859..4980
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4896..4915)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4950..4969
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5029..5045)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5029..5045)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5042..5064)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5053..5069
                     /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(5077..5083,5084..5101,5102..5107))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5122..5143
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5260..5277)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5431..6016)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5511..5530)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6187..6978,6979..7047))
                     /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     6810..6829
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7048..7152)
                     /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 attaagaata gtaaagaaac acatcataaa acctcccagg acataaaggt
     1621 gagcacagac cctgtttgga tcaagtcagt tcctggagcc tgaatgatga ctgctgaatc
     1681 acgggaagcc acgggtctgt ccccacaggc tgcacaggag aaggatggta tcgtaatagt
     1741 gaaggtggaa gaggaagatg aggaagacca catgtggggg caggattcca ccctacagga
     1801 cacgcctcct ccagacccag agatattccg ccaacgcttc aggcgcttct gttaccagaa
     1861 cacttttggg ccccgccgcg gagcgtaagt attcaaaatt ccaaaatttt ttactagaaa
     1921 tattcgattt tttaataggc agtttctata ctattgtata ctattgtaga ttcgttgaaa
     1981 agtatgtaac aggaagaata aagcatttcc gaccatgtaa agtatatata ttcttaataa
     2041 ggatcaatag ccgagtcgat ctcgccatgt ccgtctgtct tattatttta ttaccgccga
     2101 gacatcagga actataaaag ctagaaggat gagttttagc atacagattc tagagacaag
     2161 gacgcagagc aagtttgttg atccatgctg ccacgcttta actttctcaa attgcccaaa
     2221 actgccatgc ccacattttt gaactatttt cgaaattttt tcataattgt attactcgtg
     2281 taaatttcca tcaatttgcc aaaaaacttt ttgtcacgcg ttaacgccct aaagccgcca
     2341 atttggtcac gcccacacta ttgaacaatt atcaaatttt ttctcatttt attccccaat
     2401 atctatcgat atccccgatt atgaaattat taaatttcgc gttcgcattc acactagctg
     2461 agtaacgagt atctgatagt tggggaaatc gacttatttt ttatatacaa tgaaaatgaa
     2521 tttaatcata tgaatatcga ttatagcttt ttatttaata tgaatattta tttgggctta
     2581 aggtgtaacc tcctcgacat aagactcaca tggcgcaggc acattgaaga caaaaatact
     2641 cattgtcggg tctcgcaccc tccagcagca cctaaaatta tgtcttcaat tattgccaac
     2701 attggagaca caattagtct gtggcacctc aggcggccgc tcgagtctag agggcccgtt
     2761 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     2821 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     2881 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     2941 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     3001 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     3061 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     3121 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     3181 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     3241 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     3301 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     3361 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     3421 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     3481 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3541 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     3601 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     3661 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     3721 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     3781 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     3841 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     3901 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     3961 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     4021 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     4081 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     4141 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     4201 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     4261 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     4321 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     4381 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     4441 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     4501 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     4561 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     4621 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     4681 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     4741 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     4801 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     4861 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4921 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     4981 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     5041 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     5101 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     5161 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     5221 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5281 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5341 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5401 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5461 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5521 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5581 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5641 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5701 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5761 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5821 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     5881 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5941 cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag
     6001 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6061 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6121 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6181 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6241 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6301 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6361 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6421 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6481 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6541 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6601 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6661 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6721 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6781 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     6841 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6901 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6961 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7021 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7081 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7141 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
