LOCUS       sequence-97427-e        7676 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 7676)
  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 7676)
  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: pcDNA3.1(+) ZKSCAN1 Sense
FEATURES             Location/Qualifiers
     source          1..7676
                     /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(3270..3287)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3276..3500
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3546..3974
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3633..3652)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3843..3864
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3983..4003)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3988..4317
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4168..4303
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4384..5178
                     /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(4438..4457)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5048..5067
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5352..5473
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5389..5408)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5443..5462
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5522..5538)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5522..5538)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5535..5557)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5546..5562
                     /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(5570..5576,5577..5594,5595..5600))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5615..5636
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5753..5770)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5924..6509)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6004..6023)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6680..7471,7472..7540))
                     /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     7303..7322
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7541..7645)
                     /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 ggaattcact
      961 agtgattgct gtcaacctgc aggtaaatca ctctgactcc aagcaaacac cctgagggtg
     1021 gagtcggcct gtttgaggtt tctgtttgca aatcttgata tatgaagtga cagtggagat
     1081 tgtacagttt tttcctcgat ttgtcaggat tttttttttt ttgacggagt ttaacttctt
     1141 gtctcccagg taggaagtgc agtggcgtaa tctcggctca ctacaacctc cacctcctgg
     1201 gttcaagcgt ttctcctgcc tcagctttcc gagtagctgg gattacaggc gcctgccacc
     1261 atgccctgct gacttttgta tttttagtag agacggggtt tcaccatgtt ggccaggctg
     1321 gtcttgaact cctgaccgca ggcgattggc ctgcctcggc ctcccaaagt gctgagatta
     1381 caggcgtgag ccaccacccc cggcctcagg agcgttctga tagtgcctcg atgtgctgcc
     1441 tcctataaag tgttagcagc acagatcact ttttgtaaag gtacgtacta atgacttttt
     1501 ttttatactt caggaatagt aaagaaacac atcataaaac ctcccaggac ataaaggtga
     1561 gcacagaccc tgtttggatc aagtcagttc ctggagcctg aatgatgact gctgaatcac
     1621 gggaagccac gggtctgtcc ccacaggctg cacaggagaa ggatggtatc gtaatagtga
     1681 aggtggaaga ggaagatgag gaagaccaca tgtgggggca ggattccacc ctacaggaca
     1741 cgcctcctcc agacccagag atattccgcc aacgcttcag gcgcttctgt taccagaaca
     1801 cttttgggcc ccgagaggct ctcagtcggc tgaaggaact ttgtcatcag tggctgcggc
     1861 cagaaataaa caccaaggaa cagatcctgg agcttctggt gctagagcag tttctttcca
     1921 tcctgcccaa ggagctccag gtctggctgc aggaataccg ccccgatagt ggagaggagg
     1981 ccgtgaccct tctagaagac ttggagcttg atttatcagg acaacaggta aaaagaggtg
     2041 aaacctatta tgtgtgagca gggcacagac gttgaaactg gagccaggag aagtattggc
     2101 aggctttagg ttattaggtg gttactctgt cttaaaaatg ttctggcttt cttcctgcat
     2161 ccactggcat actcatggtc tgtttttaaa tattttaatt cccatttaca aagtgattta
     2221 cccacaagcc caacctgtct gtcttcaggt cccaggtcaa gttcatggac ctgagatgct
     2281 cgcaaggggg atggtgcctc tggatccagt tcaggagtcc tcgagctttg accttcatca
     2341 cgaggccacc cagtcccact tcaaacattc gtctcggaaa ccccgcctct tacagtcacg
     2401 aggtaagaag caaggtttca tttaggggaa gggaaatgat tcaggacgag agtctttgtg
     2461 ctgctgagtg cctgtgatga agaagcatgt tagtcctggg caacgtagcg agaccccatc
     2521 tctacaaaaa atagaaaaat tagccaggta tagtggcgca cacctgtgat tccagctacg
     2581 caggaggctg aggtgggagg attgcttgag cccaggaggt tgaggctgca gtgagctgga
     2641 atcatgccac tactccaacc tgggcaacac agcaaggacc ctgtctcaaa agctacttac
     2701 agaaaagaat taggctcggc acggtagctc acacctgtaa tcccagcact ttgggaggct
     2761 gaggcgggca gatcacttga ggtcaggagt ttgagaccag cctggccaac atggtgaaac
     2821 cttgtctcta ctaaaaatat gaaaattagc caggcatggt ggcacattcc tgtaatccca
     2881 gctactcggg aggctgaggc aggagaatca cttgaaccca ggaggtggag gttgcagtaa
     2941 gccgagatcg taccactgtg ctctagcctt ggtgacagag cgagactgtc ttaaaaaaaa
     3001 aaaaaaaaaa agaattaatt aaaaatttaa aaaaaaatga aaaaaagctg catgcttgtt
     3061 ttttgttttt agttattcta cattgttgtc attattacca aatattgggg aaaatacaac
     3121 ttacagacca atctcaggag ttaaatgtta ctacgaaggc aaatgaacta tgcgtaatga
     3181 acctggtagg cattaaatcg aattctgcag atatccagca cagtggcggc cgctcgagtc
     3241 tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     3301 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     3361 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     3421 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     3481 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     3541 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3601 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     3661 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     3721 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     3781 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     3841 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     3901 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     3961 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     4021 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     4081 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4141 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     4201 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     4261 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     4321 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     4381 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     4441 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     4501 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     4561 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     4621 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     4681 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     4741 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     4801 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     4861 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     4921 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     4981 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     5041 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     5101 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     5161 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     5221 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     5281 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     5341 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     5401 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     5461 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     5521 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     5581 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     5641 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     5701 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     5761 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     5821 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     5881 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     5941 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     6001 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6061 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     6121 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     6181 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6241 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     6301 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     6361 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     6421 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc
     6481 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6541 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6601 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6661 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6721 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6781 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     6841 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6901 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6961 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     7021 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     7081 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     7141 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     7201 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     7261 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     7321 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     7381 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     7441 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     7501 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     7561 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7621 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
