LOCUS       sequence-97442-e        7667 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 7667)
  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 7667)
  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-mMALAT1_3' ...
FEATURES             Location/Qualifiers
     source          1..7667
                     /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"
     primer_bind     complement(4100..4119)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4101..4119)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4126..4143)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4126..4142)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4135..4157)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4283..4711
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4370..4389)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4580..4601
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             5055..5849
                     /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(5109..5128)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5719..5738
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(5867..5935,5936..6727)
                     /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(6085..6104)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6898..7486
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7387..7406
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7640..7657
                     /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 gtcgacgatt cgtcagtagg gttgtaaagg tttttctttt
     3121 cctgagaaaa caaccttttg ttttctcagg ttttgctttt tggcctttcc ctagctttaa
     3181 aaaaaaaaaa gcaaaagacg ctggtggctg gcactcctgg tttccaggac ggggttcaag
     3241 tccctgcggt gtctttgctt gcggccgcgc tgtgcggtag gcattgaggc agccagcgca
     3301 ggggcttctg ctgagggggc aggcggagct tgaggaaacc gcagataagt ttttttctct
     3361 ttgaaagata gagattaata caactactta aaaaatatag tcaataggtt actaagatat
     3421 tgcttagcgt taagttttta acgtaatttt aatagcttaa gattttaaga gaaaatatga
     3481 agacttagaa gagtagcatg aggaaggaaa agataaaagg tttctaaaac atgacggagg
     3541 ttgagatgaa gcttcttcat ggagtaaaaa atgtatttaa aagaaaattg agagaaagga
     3601 ctacagagcc ccgaattaat accaatagaa gggcaatgct tttagattaa aatgaaggtg
     3661 acttaaacag cttaaagttt agtttaaaag ttgtaggtga ttaaaataat ttgaaggcga
     3721 tcttttaaaa agagattaaa ccgaaggtga ttaaaagacc ttgaaatcca tgacgcaggg
     3781 agaattgcgt catttaaagc ctagttaacg catttactaa acgcagacga aaatggaaag
     3841 attaattggg agtggtagga tgaaacaatt tggagaagat agaagtttga agtggaaaac
     3901 tggaagacag aagtacggga aggcgaagaa aagaatagag aagataggga aattagaaga
     3961 taaaaacata cttttagaag aaaaaagata aatttaaacc tgaaaagtag gaagcagaag
     4021 aaaaaagaca agctaggaaa caaaaagcta agggcaaaat gtacaaactc tcgagcatgc
     4081 atctagaggg cccaattcgc cctatagtga gtcgtattac aattcactgg ccgtcgtttt
     4141 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     4201 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     4261 gcgcagcctg aatggcgaat ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     4321 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     4381 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     4441 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     4501 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     4561 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     4621 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     4681 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa ttcagggcgc aagggctgct
     4741 aaaggaagcg gaacacgtag aaagccagtc cgcagaaacg gtgctgaccc cggatgaatg
     4801 tcagctactg ggctatctgg acaagggaaa acgcaagcgc aaagagaaag caggtagctt
     4861 gcagtgggct tacatggcga tagctagact gggcggtttt atggacagca agcgaaccgg
     4921 aattgccagc tggggcgccc tctggtaagg ttgggaagcc ctgcaaagta aactggatgg
     4981 ctttcttgcc gccaaggatc tgatggcgca ggggatcaag atctgatcaa gagacaggat
     5041 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     5101 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     5161 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     5221 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     5281 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     5341 aagtgccggg gcaggatctc ctgtcatccc accttgctcc tgccgagaaa gtatccatca
     5401 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     5461 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     5521 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     5581 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     5641 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     5701 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     5761 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     5821 tctatcgcct tcttgacgag ttcttctgaa ttgaaaaagg aagagtatga gtattcaaca
     5881 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     5941 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     6001 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     6061 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     6121 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     6181 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     6241 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     6301 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     6361 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     6421 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     6481 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     6541 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     6601 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     6661 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     6721 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     6781 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     6841 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     6901 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     6961 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     7021 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     7081 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     7141 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     7201 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     7261 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     7321 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     7381 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     7441 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     7501 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     7561 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     7621 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaag
//
