LOCUS       sequence-97446-e        6068 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a miniature version of the ZKSCAN1 circular RNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6068)
  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 6068)
  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(+) CircRNA Mini Vector-ZKSCAN1 nt 548...
FEATURES             Location/Qualifiers
     source          1..6068
                     /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(1662..1679)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1668..1892
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1938..2366
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2025..2044)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2235..2256
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2375..2395)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2380..2709
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2560..2695
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2622..2641
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2776..3570
                     /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(2830..2849)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3440..3459
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3744..3865
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3781..3800)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3835..3854
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3914..3930)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3914..3930)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3927..3949)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3938..3954
                     /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(3962..3968,3969..3986,3987..3992))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4007..4028
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4145..4162)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4316..4901)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4396..4415)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5072..5863,5864..5932))
                     /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     5695..5714
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5933..6037)
                     /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 ggaattcaaa
      961 gtgctgagat tacaggcgtg agccaccacc cccggcccac tttttgtaaa ggtacgtact
     1021 aatgactttt tttttatact tcaggatatc gaatagtaaa gaaacacatc ataaaacctc
     1081 ccaggacata aaggtgagca cagaccctgt ttggatcaag tcagttcctg gagcctgaat
     1141 gatgactgct gaatcacggg aagccacggg tctgtcccca caggctgcac aggagaagga
     1201 tggtatcgta atagtgaagg tggaagagga agatgaggaa gaccacatgt gggggcagga
     1261 ttccacccta caggacacgc ctcctccaga cccagagata ttccgccaac gcttcaggcg
     1321 cttctgttac cagaacactt ttgggccccg agaggctctc agtcggctga aggaactttg
     1381 tcatcagtgg ctgcggccag aaataaacac caaggaacag atcctggagc ttctggtgct
     1441 agagcagttt ctttccatcc tgcccaagga gctccaggtc tggctgcagg aataccgccc
     1501 cgatagtgga gaggaggccg tgacccttct agaagacttg gagcttgatt ccgcggaggt
     1561 aagaagcaag gaaaagaatt aggctcggca cggtagctca cacctgtaat cccagcagcg
     1621 gccgctcgag tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag
     1681 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     1741 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     1801 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     1861 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     1921 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     1981 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     2041 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     2101 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     2161 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     2221 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     2281 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     2341 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     2401 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     2461 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     2521 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     2581 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     2641 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     2701 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     2761 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     2821 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     2881 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     2941 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     3001 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     3061 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     3121 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     3181 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     3241 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     3301 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     3361 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     3421 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     3481 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     3541 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     3601 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     3661 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     3721 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     3781 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     3841 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     3901 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     3961 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     4021 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     4081 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     4141 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     4201 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     4261 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     4321 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     4381 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     4441 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     4501 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     4561 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     4621 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     4681 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     4741 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     4801 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtttttttg
     4861 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     4921 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     4981 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     5041 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     5101 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5161 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     5221 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     5281 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     5341 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     5401 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     5461 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     5521 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     5581 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     5641 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     5701 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     5761 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     5821 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     5881 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     5941 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     6001 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     6061 ctgacgtc
//
