LOCUS       sequence-97440-e        7615 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the human ZKSCAN1 exon 2/3 circular RNA .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7615)
  AUTHORS   Liang D, Wilusz JE
  TITLE     Short intronic repeat sequences facilitate circular RNA production.
  JOURNAL   Genes Dev. 2014 Oct 3. pii: gad.251926.114.
  PUBMED    25281217
REFERENCE   2  (bases 1 to 7615)
  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. 
            2014 Oct 3. pii: gad.251926.114."
COMMENT     Sequence Label: pCRII-TOPO CMV-ZKSCAN1 nt 400-1782-SV40 pA Sense
FEATURES             Location/Qualifiers
     source          1..7615
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        239..257
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     239..256
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        1075..1453
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1454..1657
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1608..1628
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    3087..3208
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3124..3143)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3178..3197
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4048..4067)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4049..4067)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4074..4091)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4074..4090)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4083..4105)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4231..4659
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4318..4337)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4528..4549
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             5003..5797
                     /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(5057..5076)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5667..5686
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(5815..5883,5884..6675)
                     /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     complement(6033..6052)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6846..7434
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7335..7354
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7588..7605
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctat
      241 ttaggtgaca ctatagaata ctcaagctat gcatcaagct tctttaaaag gccacttgaa
      301 ctcgctttcc atggcgattt gccttgtgag cactttcagg agagcctgga agctgaaaaa
      361 cggtagaaaa atttccgtgc gggccgtggg gggctggcgg caactggggg gccgcagatc
      421 agagtgggcc actggcagcc aacggccccc ggggctcagg cggggagcag ctctgtggtg
      481 tgggattgag gcgttttcca agagtgggtt ttcacgtttc taagatttcc caagcagaca
      541 gcccgtgctg ctccgatttc tcgaacaaaa aagcaaaacg tgtggctgtc ttgggagcaa
      601 gtcgcaggac tgcaagcagt tgggggagaa agtccgccat tttgccactt ctcaaccgtc
      661 cctgcaaggc tggggctcag ttgcgtaatg gaaagtaaag ccctgaacta tcacacttta
      721 atcttccttc aaaaggtggt aaactatacc tactgtccct caagagaaca caagaagtgc
      781 tttaagaggt attttaaaag ttccgggggt tttgtgaggt gtttgatgac ccgtttaaaa
      841 tatgatttcc atgtttcttt tgtctaaagt ttgcagctca aatctttcca cacgctagta
      901 atttaagtat ttctgcatgt gtagtttgca ttcaagttcc ataagctgtt aagaaaaatc
      961 tagaaaagta aaactagaac ctatttttaa ccgaagaact actttttgcc tccctcacaa
     1021 aggcggcgga aggtgatcga attccggtga tgcgagttgt tctccgtcgg atccacattg
     1081 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
     1141 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     1201 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     1261 ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac atcaagtgta
     1321 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     1381 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
     1441 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
     1501 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     1561 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
     1621 taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta gagaacccac
     1681 tgcttactgg cgatatcaaa gtgctgagat tacaggcgtg agccaccacc cccggcctca
     1741 ggagcgttct gatagtgcct cgatgtgctg cctcctataa agtgttagca gcacagatca
     1801 ctttttgtaa aggtacgtac taatgacttt ttttttatac ttcaggaata gtaaagaaac
     1861 acatcataaa acctcccagg acataaaggt gagcacagac cctgtttgga tcaagtcagt
     1921 tcctggagcc tgaatgatga ctgctgaatc acgggaagcc acgggtctgt ccccacaggc
     1981 tgcacaggag aaggatggta tcgtaatagt gaaggtggaa gaggaagatg aggaagacca
     2041 catgtggggg caggattcca ccctacagga cacgcctcct ccagacccag agatattccg
     2101 ccaacgcttc aggcgcttct gttaccagaa cacttttggg ccccgagagg ctctcagtcg
     2161 gctgaaggaa ctttgtcatc agtggctgcg gccagaaata aacaccaagg aacagatcct
     2221 ggagcttctg gtgctagagc agtttctttc catcctgccc aaggagctcc aggtctggct
     2281 gcaggaatac cgccccgata gtggagagga ggccgtgacc cttctagaag acttggagct
     2341 tgatttatca ggacaacagg taaaaagagg tgaaacctat tatgtgtgag cagggcacag
     2401 acgttgaaac tggagccagg agaagtattg gcaggcttta ggttattagg tggttactct
     2461 gtcttaaaaa tgttctggct ttcttcctgc atccactggc atactcatgg tctgttttta
     2521 aatattttaa ttcccattta caaagtgatt tacccacaag cccaacctgt ctgtcttcag
     2581 gtcccaggtc aagttcatgg acctgagatg ctcgcaaggg ggatggtgcc tctggatcca
     2641 gttcaggagt cctcgagcct tgaccttcat cacgaggcca cccagtccca cttcaaacat
     2701 tcgtctcgga aaccccgcct cttacagtca cgaggtaaga agcaaggttt catttagggg
     2761 aagggaaatg attcaggacg agagtctttg tgctgctgag tgcctgtgat gaagaagcat
     2821 gttagtcctg ggcaacgtag cgagacccca tctctacaaa aaatagaaaa attagccagg
     2881 tatagtggcg cacacctgtg attccagcta cgcaggaggc tgaggtggga ggattgcttg
     2941 agcccaggag gttgaggctg cagtgagctg gaatcatgcc actactccaa cctgggcaac
     3001 acagcaagga ccctgtctca aaagctactt acagaaaaga attaggctcg gcacggtagc
     3061 tcacacctgt aatcccagca gtcgacaact tgtttattgc agcttataat ggttacaaat
     3121 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     3181 gtttgtccaa actcatcaat gtatcttagc ggccgcgctg tgcggtaggc attgaggcag
     3241 ccagcgcagg ggcttctgct gagggggcag gcggagcttg aggaaaccgc agataagttt
     3301 ttttctcttt gaaagataga gattaataca actacttaaa aaatatagtc aataggttac
     3361 taagatattg cttagcgtta agtttttaac gtaattttaa tagcttaaga ttttaagaga
     3421 aaatatgaag acttagaaga gtagcatgag gaaggaaaag ataaaaggtt tctaaaacat
     3481 gacggaggtt gagatgaagc ttcttcatgg agtaaaaaat gtatttaaaa gaaaattgag
     3541 agaaaggact acagagcccc gaattaatac caatagaagg gcaatgcttt tagattaaaa
     3601 tgaaggtgac ttaaacagct taaagtttag tttaaaagtt gtaggtgatt aaaataattt
     3661 gaaggcgatc ttttaaaaag agattaaacc gaaggtgatt aaaagacctt gaaatccatg
     3721 acgcagggag aattgcgtca tttaaagcct agttaacgca tttactaaac gcagacgaaa
     3781 atggaaagat taattgggag tggtaggatg aaacaatttg gagaagatag aagtttgaag
     3841 tggaaaactg gaagacagaa gtacgggaag gcgaagaaaa gaatagagaa gatagggaaa
     3901 ttagaagata aaaacatact tttagaagaa aaaagataaa tttaaacctg aaaagtagga
     3961 agcagaagaa aaaagacaag ctaggaaaca aaaagctaag ggcaaaatgt acaaactctc
     4021 gagcatgcat ctagagggcc caattcgccc tatagtgagt cgtattacaa ttcactggcc
     4081 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     4141 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     4201 caacagttgc gcagcctgaa tggcgaatgg acgcgccctg tagcggcgca ttaagcgcgg
     4261 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     4321 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     4381 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     4441 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     4501 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     4561 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     4621 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaatt cagggcgcaa
     4681 gggctgctaa aggaagcgga acacgtagaa agccagtccg cagaaacggt gctgaccccg
     4741 gatgaatgtc agctactggg ctatctggac aagggaaaac gcaagcgcaa agagaaagca
     4801 ggtagcttgc agtgggctta catggcgata gctagactgg gcggttttat ggacagcaag
     4861 cgaaccggaa ttgccagctg gggcgccctc tggtaaggtt gggaagccct gcaaagtaaa
     4921 ctggatggct ttcttgccgc caaggatctg atggcgcagg ggatcaagat ctgatcaaga
     4981 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     5041 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     5101 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     5161 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     5221 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     5281 attgggcgaa gtgccggggc aggatctcct gtcatcccac cttgctcctg ccgagaaagt
     5341 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     5401 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     5461 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     5521 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     5581 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     5641 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     5701 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     5761 catcgccttc tatcgccttc ttgacgagtt cttctgaatt gaaaaaggaa gagtatgagt
     5821 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     5881 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     5941 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     6001 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     6061 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     6121 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     6181 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     6241 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     6301 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     6361 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     6421 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     6481 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     6541 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     6601 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     6661 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     6721 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     6781 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     6841 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     6901 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     6961 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     7021 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     7081 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     7141 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     7201 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     7261 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     7321 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     7381 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     7441 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     7501 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     7561 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaag
//
