LOCUS       sequence-127146-        7314 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 7314)
  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 7314)
  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(+) ZKSCAN1 MCS-ZKSCAN1 548-1417 (No i...
FEATURES             Location/Qualifiers
     source          1..7314
                     /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(2908..2925)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2914..3138
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3184..3612
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3271..3290)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3481..3502
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3621..3641)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3626..3955
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3806..3941
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4022..4816
                     /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(4076..4095)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4686..4705
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4990..5111
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5027..5046)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5081..5100
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5160..5176)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5160..5176)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5173..5195)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5184..5200
                     /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(5208..5214,5215..5232,5233..5238))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5253..5274
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5391..5408)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5562..6147)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5642..5661)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6318..7109,7110..7178))
                     /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     6941..6960
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7179..7283)
                     /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 ggaattcagt
      961 gacagtggag attgtacagt tttttcctcg atttgtcagg attttttttt tttgacggag
     1021 tttaacttct tgtctcccag gtaggaagtg cagtggcgta atctcggctc actacaacct
     1081 ccacctcctg ggttcaagcg tttctcctgc ctcagctttc cgagtagctg ggattacagg
     1141 cgcctgccac catgccctgc tgacttttgt atttttagta gagacggggt ttcaccatgt
     1201 tggccaggct ggtcttgaac tcctgaccgc aggcgattgg cctgcctcgg cctcccaaag
     1261 tgctgagatt acaggcgtga gccaccaccc ccggcctcag gagcgttctg atagtgcctc
     1321 gatgtgctgc ctcctataaa gtgttagcag cacagatcac tttttgtaaa ggtacgtact
     1381 aatgactttt tttttatact tcaggatatc gaatagtaaa gaaacacatc ataaaacctc
     1441 ccaggacata aaggtgagca cagaccctgt ttggatcaag tcagttcctg gagcctgaat
     1501 gatgactgct gaatcacggg aagccacggg tctgtcccca caggctgcac aggagaagga
     1561 tggtatcgta atagtgaagg tggaagagga agatgaggaa gaccacatgt gggggcagga
     1621 ttccacccta caggacacgc ctcctccaga cccagagata ttccgccaac gcttcaggcg
     1681 cttctgttac cagaacactt ttgggccccg agaggctctc agtcggctga aggaactttg
     1741 tcatcagtgg ctgcggccag aaataaacac caaggaacag atcctggagc ttctggtgct
     1801 agagcagttt ctttccatcc tgcccaagga gctccaggtc tggctgcagg aataccgccc
     1861 cgatagtgga gaggaggccg tgacccttct agaagacttg gagcttgatt tatcaggaca
     1921 acaggtccca ggtcaagttc atggacctga gatgctcgca agggggatgg tgcctctgga
     1981 tccagttcag gagtcctcga gccttgacct tcatcacgag gccacccagt cccacttcaa
     2041 acattcgtct cggaaacccc cgcggaggta agaagcaagg tttcatttag gggaagggaa
     2101 atgattcagg acgagagtct ttgtgctgct gagtgcctgt gatgaagaag catgttagtc
     2161 ctgggcaacg tagcgagacc ccatctctac aaaaaataga aaaattagcc aggtatagtg
     2221 gcgcacacct gtgattccag ctacgcagga ggctgaggtg ggaggattgc ttgagcccag
     2281 gaggttgagg ctgcagtgag ctgtaatcat gccactactc caacctgggc aacacagcaa
     2341 ggaccctgtc tcaaaagcta cttacagaaa agaattaggc tcggcacggt agctcacacc
     2401 tgtaatccca gcactttggg aggctgaggc gggcagatca cttgaggtca ggagtttgag
     2461 accagcctgg ccaacatggt gaaaccttgt ctctactaaa aatatgaaaa ttagccaggc
     2521 atggtggcac attcctgtaa tcccagctac tcgggaggct gaggcaggag aatcacttga
     2581 acccaggagg tggaggttgc agtaagccga gatcgtacca ctgtgctcta gccttggtga
     2641 cagagcgaga ctgtcttaaa aaaaaaaaaa aaaaaaaaag aattaattaa aaatttaaaa
     2701 aaaaatgaaa aaaagctgca tgcttgtttt ttgtttttag ttattctaca ttgttgtcat
     2761 tattaccaaa tattggggaa aatacaactt acagaccaat ctcaggagtt aaatgttact
     2821 acgaaggcaa atgaactatg cgtaatgaac ctggtaggca ttagcggccg ctcgagtcta
     2881 gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     2941 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     3001 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     3061 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     3121 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc
     3181 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     3241 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     3301 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     3361 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     3421 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     3481 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     3541 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     3601 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     3661 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3721 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3781 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3841 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     3901 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     3961 cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg
     4021 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     4081 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     4141 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     4201 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     4261 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     4321 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     4381 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     4441 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     4501 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     4561 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     4621 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     4681 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     4741 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     4801 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     4861 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     4921 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     4981 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     5041 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     5101 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     5161 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     5221 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     5281 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     5341 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     5401 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     5461 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     5521 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5581 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5641 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5701 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     5761 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5821 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5881 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5941 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     6001 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     6061 agctcttgat ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg caagcagcag
     6121 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6181 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6241 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     6301 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     6361 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     6421 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6481 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6541 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6601 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6661 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6721 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6781 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6841 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6901 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6961 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     7021 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     7081 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     7141 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     7201 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     7261 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
