LOCUS       sequence-127059-        7353 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Drosophila laccase2 exon 2 circular RNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7353)
  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 7353)
  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 Sense
FEATURES             Location/Qualifiers
     source          1..7353
                     /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(2947..2964)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2953..3177
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3223..3651
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3310..3329)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3520..3541
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3660..3680)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3665..3994
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3845..3980
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3907..3926
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4061..4855
                     /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(4115..4134)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4725..4744
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5029..5150
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5066..5085)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5120..5139
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5199..5215)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5199..5215)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5212..5234)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5223..5239
                     /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(5247..5253,5254..5271,5272..5277))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5292..5313
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5430..5447)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5601..6186)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5681..5700)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6357..7148,7149..7217))
                     /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     6980..6999
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7218..7322)
                     /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 gcagaaaatc aaccagatct tacaccaatg gggtccttct ggcagcaaca tgtggccact
     1621 cctagtagtt caactccatt cgattcacca tttttttcgg caacacacgg ccttgtgcag
     1681 actcacccgt ctataggtgg aagccactct gggcccgctc cattaaatgg caatagcctt
     1741 gggtctggag ctccatcctc aatcctagct ggtactaagc cagccctaac cgctggaagc
     1801 ggttacctaa actcacgcgg gagtctaccg acctctgcac gttatccagc taataagatc
     1861 acaggaaccg tcgtggaacc caacccaaaa tctccattcc gccacttaga tttctctact
     1921 agtgcaactg ccgagctgcg gcgcaatcct gcgctatctg ctcctgatga atgtgcacgc
     1981 gcttgtcgcg aaggcgagcc tccgagaatt tgctactatc attttacgtt ggaatactac
     2041 acagtattgg gagcgtaagt attcaaaatt ccaaaatttt ttactagaaa tattcgattt
     2101 tttaataggc agtttctata ctattgtata ctattgtaga ttcgttgaaa agtatgtaac
     2161 aggaagaata aagcatttcc gaccatgtaa agtatatata ttcttaataa ggatcaatag
     2221 ccgagtcgat ctcgccatgt ccgtctgtct tattgtttta ttaccgccga gacatcagga
     2281 actataaaag ctagaaggat gagttttagc atacagattc tagagacaag gacgcagagc
     2341 aagtttgttg atccatgctg ccacgcttta actttctcaa attgcccaaa actgccatgc
     2401 ccacattttt gaactatttt cgaaattttt tcataattgt attactcgtg taaatttcca
     2461 tcaatttgcc aaaaaacttt ttgtcacgcg ttaacgccct aaagccgcca atttggtcac
     2521 gcccacacta ttgagcaatt atcaaatttt ttctcatttt attccccaat atctatcgat
     2581 atccccgatt atgaaattat taaatttcgc gttcgcattc acactagctg agtaacgagt
     2641 atctgatagt tggggaaatc gacttatttt ttatatacaa tgaaaatgaa tttaatcata
     2701 tgaatatcga ttatagcttt ttatttaata tgaatattta tttgggctta aggtgtaacc
     2761 tcctcgacat aagactcaca tggcgcaggc acattgaaga caaaaatact cattgtcggg
     2821 tctcgcaccc tccagcagca cctaaaatta tgtcttcaat tattgccaac attggagaca
     2881 caattagtct gtggcacctc aggcggccgc tcgagtctag agggcccgtt taaacccgct
     2941 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     3001 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     3061 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     3121 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     3181 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     3241 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     3301 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     3361 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     3421 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     3481 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     3541 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     3601 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     3661 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3721 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     3781 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3841 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3901 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     3961 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     4021 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     4081 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     4141 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     4201 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     4261 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     4321 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     4381 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     4441 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     4501 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     4561 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     4621 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     4681 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     4741 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     4801 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     4861 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     4921 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     4981 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     5041 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     5101 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     5161 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     5221 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     5281 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     5341 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     5401 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     5461 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     5521 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     5581 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5641 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5701 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5761 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     5821 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5881 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5941 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     6001 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     6061 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     6121 accaccgctg gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6181 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6241 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6301 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     6361 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     6421 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     6481 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     6541 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     6601 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     6661 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     6721 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     6781 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     6841 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     6901 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     6961 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     7021 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     7081 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     7141 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     7201 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     7261 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     7321 cgcacatttc cccgaaaagt gccacctgac gtc
//
