LOCUS       sequence-97433-e        6443 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ZKSCAN1 circular RNA expression plasmid containing minimal 
            sufficient introns.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6443)
  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 6443)
  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 nt 400-1782 delta440-500
            d...
FEATURES             Location/Qualifiers
     source          1..6443
                     /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(2037..2054)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2043..2267
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2313..2741
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2400..2419)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2610..2631
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2750..2770)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2755..3084
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2935..3070
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2997..3016
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3151..3945
                     /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(3205..3224)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3815..3834
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4119..4240
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4156..4175)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4210..4229
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4289..4305)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4289..4305)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4302..4324)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4313..4329
                     /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(4337..4343,4344..4361,4362..4367))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4382..4403
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4520..4537)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4691..5276)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4771..4790)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5447..6238,6239..6307))
                     /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     6070..6089
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6308..6412)
                     /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 tcaggaatag taaagaaaca catcataaaa cctcccagga
     1081 cataaaggtg agcacagacc ctgtttggat caagtcagtt cctggagcct gaatgatgac
     1141 tgctgaatca cgggaagcca cgggtctgtc cccacaggct gcacaggaga aggatggtat
     1201 cgtaatagtg aaggtggaag aggaagatga ggaagaccac atgtgggggc aggattccac
     1261 cctacaggac acgcctcctc cagacccaga gatattccgc caacgcttca ggcgcttctg
     1321 ttaccagaac acttttgggc cccgagaggc tctcagtcgg ctgaaggaac tttgtcatca
     1381 gtggctgcgg ccagaaataa acaccaagga acagatcctg gagcttctgg tgctagagca
     1441 gtttctttcc atcctgccca aggagctcca ggtctggctg caggaatacc gccccgatag
     1501 tggagaggag gccgtgaccc ttctagaaga cttggagctt gatttatcag gacaacaggt
     1561 aaaaagaggt gaaacctatt atgtgtgagc agggcacaga cgttgaaact ggagccagga
     1621 gaagtattgg caggctttag gttattaggt ggttactctg tcttaaaaat gttctggctt
     1681 tcttcctgca tccactggca tactcatggt ctgtttttaa atattttaat tcccatttac
     1741 aaagtgattt acccacaagc ccaacctgtc tgtcttcagg tcccaggtca agttcatgga
     1801 cctgagatgc tcgcaagggg gatggtgcct ctggatccag ttcaggagtc ctcgagcctt
     1861 gaccttcatc acgaggccac ccagtcccac ttcaaacatt cgtctcggaa accccgcctc
     1921 ttacagtcac gaggtaagaa gcaaggaaaa gaattaggct cggcacggta gctcacacct
     1981 gtaatcccag cagcggccgc tcgagtctag agggcccgtt taaacccgct gatcagcctc
     2041 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     2101 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     2161 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     2221 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     2281 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     2341 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     2401 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2461 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     2521 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     2581 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     2641 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     2701 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     2761 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     2821 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     2881 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     2941 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     3001 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     3061 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     3121 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     3181 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     3241 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     3301 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     3361 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     3421 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     3481 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     3541 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     3601 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     3661 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     3721 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     3781 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     3841 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     3901 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     3961 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     4021 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     4081 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     4141 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4201 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     4261 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     4321 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     4381 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     4441 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     4501 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4561 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4621 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4681 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4741 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4801 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4861 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4921 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4981 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     5041 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     5101 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     5161 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5221 gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5281 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5341 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5401 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5461 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5521 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5581 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5641 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5701 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5761 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5821 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5881 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5941 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6001 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6061 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6121 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6181 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6241 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6301 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6361 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6421 cccgaaaagt gccacctgac gtc
//
