LOCUS       sequence-127117-        7383 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 7383)
  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 7383)
  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 548-1047
FEATURES             Location/Qualifiers
     source          1..7383
                     /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(2977..2994)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2983..3207
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3253..3681
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3340..3359)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3550..3571
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3690..3710)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3695..4024
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3875..4010
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3937..3956
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4091..4885
                     /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(4145..4164)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4755..4774
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5059..5180
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5096..5115)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5150..5169
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5229..5245)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5229..5245)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5242..5264)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5253..5269
                     /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(5277..5283,5284..5301,5302..5307))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5322..5343
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5460..5477)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5631..6216)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5711..5730)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6387..7178,7179..7247))
                     /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     7010..7029
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7248..7352)
                     /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 ccccgagagg ctctcagtcg gctgaaggaa ctttgtcatc agtggctgcg
     1921 gccagaaata aacaccaagg aacagatcct ggagcttctg gtgctagagc agtttctttc
     1981 catcctgccc aaggagctcc aggtctggct gcaggaatac cgccccgata gtggagagga
     2041 ggccgtgacc cttctagaag acttggagct tgattccgcg gagcgtaagt attcaaaatt
     2101 ccaaaatttt ttactagaaa tattcgattt tttaataggc agtttctata ctattgtata
     2161 ctattgtaga ttcgttgaaa agtatgtaac aggaagaata aagcatttcc gaccatgtaa
     2221 agtatatata ttcttaataa ggatcaatag ccgagtcgat ctcgccatgt ccgtctgtct
     2281 tattatttta ttaccgccga gacatcagga actataaaag ctagaaggat gagttttagc
     2341 atacagattc tagagacaag gacgcagagc aagtttgttg atccatgctg ccacgcttta
     2401 actttctcaa attgcccaaa actgccatgc ccacattttt gaactatttt cgaaattttt
     2461 tcataattgt attactcgtg taaatttcca tcaatttgcc aaaaaacttt ttgtcacgcg
     2521 ttaacgccct aaagccgcca atttggtcac gcccacacta ttgaacaatt atcaaatttt
     2581 ttctcatttt attccccaat atctatcgat atccccgatt atgaaattat taaatttcgc
     2641 gttcgcattc acactagctg agtaacgagt atctgatagt tggggaaatc gacttatttt
     2701 ttatatacaa tgaaaatgaa tttaatcata tgaatatcga ttatagcttt ttatttaata
     2761 tgaatattta tttgggctta aggtgtaacc tcctcgacat aagactcaca tggcgcaggc
     2821 acattgaaga caaaaatact cattgtcggg tctcgcaccc tccagcagca cctaaaatta
     2881 tgtcttcaat tattgccaac attggagaca caattagtct gtggcacctc aggcggccgc
     2941 tcgagtctag agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc
     3001 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     3061 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     3121 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     3181 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     3241 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     3301 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     3361 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     3421 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     3481 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     3541 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     3601 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     3661 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     3721 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     3781 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3841 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     3901 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     3961 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     4021 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg
     4081 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     4141 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     4201 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     4261 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     4321 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     4381 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     4441 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     4501 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     4561 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     4621 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     4681 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     4741 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     4801 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     4861 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     4921 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     4981 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     5041 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     5101 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     5161 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     5221 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     5281 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     5341 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     5401 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     5461 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     5521 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     5581 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     5641 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5701 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     5761 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     5821 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5881 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5941 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     6001 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     6061 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     6121 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggttt ttttgtttgc
     6181 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     6241 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     6301 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     6361 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6421 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6481 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6541 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6601 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6661 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6721 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6781 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6841 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6901 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6961 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     7021 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     7081 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     7141 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     7201 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     7261 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7321 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     7381 gtc
//
