LOCUS       sequence-127126-        7981 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the human HIPK3 exon 2 circular RNA in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7981)
  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 7981)
  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(+) Laccase2 MCS-HIPK3 Exon
FEATURES             Location/Qualifiers
     source          1..7981
                     /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(3575..3592)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3581..3805
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3851..4279
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3938..3957)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4148..4169
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4288..4308)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4293..4622
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4473..4608
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4535..4554
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4689..5483
                     /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(4743..4762)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5353..5372
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5657..5778
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5694..5713)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5748..5767
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5827..5843)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5827..5843)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5840..5862)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5851..5867
                     /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(5875..5881,5882..5899,5900..5905))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5920..5941
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6058..6075)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6229..6814)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6309..6328)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6985..7776,7777..7845))
                     /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     7608..7627
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7846..7950)
                     /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 ggaattccat
      961 tgagaaatga ctgagttccg gtgctctcaa gtcattgatc tttgtcgact tttatttggt
     1021 ctctgtaata acgacttcaa aaacattaaa ttctgttgcg aagccagtaa gctacaaaaa
     1081 gaaaaaacaa gagagaatgc tatagtcgta tagtatagtt tcccgactat ctgataccca
     1141 ttacttatct agggggaatg cgaacccaaa attttatcag ttttctcgga tatcgataga
     1201 tattggggaa taaatttaaa taaataaatt ttgggcgggt ttagggcgtg gcaaaaagtt
     1261 ttttggcaaa tcgctagaaa tttacaagac ttataaaatt atgaaaaaat acaacaaaat
     1321 tttaaacacg tgggcgtgac agttttgggc ggttttaggg cgttagagta ggcgaggaca
     1381 gggttacatc gactaggctt tgatcctgat caagaatata tatactttat accgcttcct
     1441 tctacatgtt acctattttt caacgaatct agtatacctt tttactgtac gatttatggg
     1501 tataataata agctaaatcg agactaagtt ttattgttat atatattttt tttattttat
     1561 gcagaaatta attaagtatg gcctcacaag tcttggtcta cccaccatat gtttatcaaa
     1621 ctcagtcaag tgccttttgt agtgtgaaga aactcaaagt agagccaagc agttgtgtat
     1681 tccaggaaag aaactatcca cggacctatg tgaatggtag aaactttgga aattctcatc
     1741 ctcccactaa gggtagtgct tttcagacaa agataccatt taatagacct cgaggacaca
     1801 acttttcatt gcagacaagt gctgttgttt tgaaaaacac tgcaggtgct acaaaggtca
     1861 tagcagctca ggcacagcaa gctcacgtgc aggcacctca gattggggcg tggcgaaaca
     1921 gattgcattt cctagaaggc ccccagcgat gtggattgaa gcgcaagagt gaggagttgg
     1981 ataatcatag cagcgcaatg cagattgtcg atgaattgtc catacttcct gcaatgttgc
     2041 aaaccaacat gggaaatcca gtgacagttg tgacagctac cacaggatca aaacagaatt
     2101 gtaccactgg agaaggtgac tatcagttag tacagcatga agtcttatgc tccatgaaaa
     2161 tacttacgaa gtccttgatt ttcttggtcg aggcacgttt ggccaggtag ttaaatgctg
     2221 gaaaagaggg acaaatgaaa ttgtagcaat caaaattttg aagaatcatc cttcttatgc
     2281 ccgtcaaggt caaatagaag tgagcatatt agcaaggctc agtactgaaa atgctgatga
     2341 atataacttt gtacgagctt atgaatgctt tcagcaccgt aaccatactt gtttagtctt
     2401 tgagatgctg gaacaaaact tgtatgactt tctgaaacaa aataaattta gtcccctgcc
     2461 actaaaagtg attcggccca ttcttcaaca agtggccact gcactgaaaa aattgaaaag
     2521 tcttggttta attcatgctg atctcaagcc agagaatatt atgttggtgg atcctgttcg
     2581 gcagccttac agggttaaag taatagactt tgggtcggcc agtcatgtat caaagactgt
     2641 ttgttcaaca tatctacaat ctcggtacta cagccgcgga gcgtaagtat tcaaaattcc
     2701 aaaatttttt actagaaata ttcgattttt taataggcag tttctatact attgtatact
     2761 attgtagatt cgttgaaaag tatgtaacag gaagaataaa gcatttccga ccatgtaaag
     2821 tatatatatt cttaataagg atcaatagcc gagtcgatct cgccatgtcc gtctgtctta
     2881 ttattttatt accgccgaga catcaggaac tataaaagct agaaggatga gttttagcat
     2941 acagattcta gagacaagga cgcagagcaa gtttgttgat ccatgctgcc acgctttaac
     3001 tttctcaaat tgcccaaaac tgccatgccc acatttttga actattttcg aaattttttc
     3061 ataattgtat tactcgtgta aatttccatc aatttgccaa aaaacttttt gtcacgcgtt
     3121 aacgccctaa agccgccaat ttggtcacgc ccacactatt gaacaattat caaatttttt
     3181 ctcattttat tccccaatat ctatcgatat ccccgattat gaaattatta aatttcgcgt
     3241 tcgcattcac actagctgag taacgagtat ctgatagttg gggaaatcga cttatttttt
     3301 atatacaatg aaaatgaatt taatcatatg aatatcgatt atagcttttt atttaatatg
     3361 aatatttatt tgggcttaag gtgtaacctc ctcgacataa gactcacatg gcgcaggcac
     3421 attgaagaca aaaatactca ttgtcgggtc tcgcaccctc cagcagcacc taaaattatg
     3481 tcttcaatta ttgccaacat tggagacaca attagtctgt ggcacctcag gcggccgctc
     3541 gagtctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag
     3601 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     3661 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     3721 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     3781 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
     3841 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     3901 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     3961 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     4021 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     4081 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     4141 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     4201 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     4261 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     4321 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     4381 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4441 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     4501 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     4561 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     4621 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat
     4681 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     4741 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     4801 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     4861 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     4921 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     4981 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     5041 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     5101 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     5161 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     5221 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     5281 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     5341 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     5401 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     5461 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     5521 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     5581 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     5641 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     5701 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     5761 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     5821 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     5881 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     5941 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     6001 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     6061 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     6121 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     6181 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     6241 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     6301 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     6361 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     6421 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     6481 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     6541 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     6601 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     6661 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     6721 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggttttt ttgtttgcaa
     6781 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     6841 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     6901 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     6961 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     7021 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     7081 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     7141 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     7201 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     7261 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     7321 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     7381 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     7441 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     7501 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     7561 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     7621 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     7681 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     7741 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     7801 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     7861 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     7921 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     7981 c
//
