LOCUS       sequence-97441-e        7721 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 7721)
  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 7721)
  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-bGH pA Sense
FEATURES             Location/Qualifiers
     source          1..7721
                     /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"
     polyA_signal    3090..3314
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4154..4173)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4155..4173)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4180..4197)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4180..4196)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4189..4211)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4337..4765
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4424..4443)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4634..4655
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             5109..5903
                     /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(5163..5182)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5773..5792
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(5921..5989,5990..6781)
                     /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(6139..6158)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6952..7540
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7441..7460
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7694..7711
                     /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 gtcgaccgac tgtgccttct agttgccagc catctgttgt
     3121 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     3181 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     3241 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     3301 ggtgggctct atgggcggcc gcgctgtgcg gtaggcattg aggcagccag cgcaggggct
     3361 tctgctgagg gggcaggcgg agcttgagga aaccgcagat aagttttttt ctctttgaaa
     3421 gatagagatt aatacaacta cttaaaaaat atagtcaata ggttactaag atattgctta
     3481 gcgttaagtt tttaacgtaa ttttaatagc ttaagatttt aagagaaaat atgaagactt
     3541 agaagagtag catgaggaag gaaaagataa aaggtttcta aaacatgacg gaggttgaga
     3601 tgaagcttct tcatggagta aaaaatgtat ttaaaagaaa attgagagaa aggactacag
     3661 agccccgaat taataccaat agaagggcaa tgcttttaga ttaaaatgaa ggtgacttaa
     3721 acagcttaaa gtttagttta aaagttgtag gtgattaaaa taatttgaag gcgatctttt
     3781 aaaaagagat taaaccgaag gtgattaaaa gaccttgaaa tccatgacgc agggagaatt
     3841 gcgtcattta aagcctagtt aacgcattta ctaaacgcag acgaaaatgg aaagattaat
     3901 tgggagtggt aggatgaaac aatttggaga agatagaagt ttgaagtgga aaactggaag
     3961 acagaagtac gggaaggcga agaaaagaat agagaagata gggaaattag aagataaaaa
     4021 catactttta gaagaaaaaa gataaattta aacctgaaaa gtaggaagca gaagaaaaaa
     4081 gacaagctag gaaacaaaaa gctaagggca aaatgtacaa actctcgagc atgcatctag
     4141 agggcccaat tcgccctata gtgagtcgta ttacaattca ctggccgtcg ttttacaacg
     4201 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     4261 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     4321 cctgaatggc gaatggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     4381 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     4441 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     4501 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     4561 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     4621 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     4681 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     4741 atttaacaaa aatttaacgc gaattttaac aaaattcagg gcgcaagggc tgctaaagga
     4801 agcggaacac gtagaaagcc agtccgcaga aacggtgctg accccggatg aatgtcagct
     4861 actgggctat ctggacaagg gaaaacgcaa gcgcaaagag aaagcaggta gcttgcagtg
     4921 ggcttacatg gcgatagcta gactgggcgg ttttatggac agcaagcgaa ccggaattgc
     4981 cagctggggc gccctctggt aaggttggga agccctgcaa agtaaactgg atggctttct
     5041 tgccgccaag gatctgatgg cgcaggggat caagatctga tcaagagaca ggatgaggat
     5101 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     5161 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     5221 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     5281 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     5341 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     5401 cggggcagga tctcctgtca tcccaccttg ctcctgccga gaaagtatcc atcatggctg
     5461 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     5521 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     5581 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     5641 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     5701 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     5761 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     5821 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     5881 gccttcttga cgagttcttc tgaattgaaa aaggaagagt atgagtattc aacatttccg
     5941 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     6001 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     6061 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     6121 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     6181 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     6241 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     6301 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     6361 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     6421 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     6481 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     6541 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     6601 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     6661 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     6721 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     6781 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     6841 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     6901 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     6961 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     7021 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     7081 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     7141 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     7201 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     7261 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     7321 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     7381 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     7441 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     7501 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     7561 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     7621 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     7681 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa g
//
