LOCUS       sequence-358804-        4770 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses GFP CDS-derived linear and circular RNA transcript. 
            Flanking introns contain NOVA2 binding sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4770)
  AUTHORS   Knupp D, Cooper DA, Saito Y, Darnell RB, Miura P
  TITLE     NOVA2 regulates neural circRNA biogenesis.
  JOURNAL   Nucleic Acids Res. 2021 Jul 9;49(12):6849-6862. doi: 
            10.1093/nar/gkab523.
  PUBMED    34157123
REFERENCE   2  (bases 1 to 4770)
  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"; doi: "10"; journalName: 
            "Nucleic Acids Res"; date: "2021-07-9- 9"; volume: "49"; issue: 
            "12"; pages: "6849-6862"
COMMENT     Sequence Label: pMini-Both
FEATURES             Location/Qualifiers
     source          1..4770
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     118..137
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        371..750
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        751..954
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     905..925
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     complement(990..1018)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2355..2375
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     promoter        complement(2466..2484)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2467..2484)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2504..2521)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2510..2734
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2816..2835)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2881..2936
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2935..2954)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     promoter        3263..3367
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3368..3436,3437..4228)
                     /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(3586..3605)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4399..4770,1..217)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
       61 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
      121 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
      181 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
      241 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
      301 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
      361 gaccgagcgc gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
      421 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
      481 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      541 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      601 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      661 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      721 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      781 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      841 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      901 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctgctgga
      961 tgagtaaagg agaagaactt tttactggag ttgtcccaat tcttgttgaa ttagatggtg
     1021 atgttaatgg gcacaaattt tctgtcagtg gtaagtccgg cgtccggggc gtcgtgccct
     1081 ggccgcctct gccttttttt tgacggagtt taacttcttg tctccaacgc acgaagtgca
     1141 gtggcgtaat ctcggctaac tacaacctca acctcctggg ttcaagcgtt tctcctgcct
     1201 aacctttctg ggattacacg cgcctgcaac aatgccctgc tgacttttgt atttttacta
     1261 cacacggggt ttaacaatgt ttttatatta actgtgattt tttgtcctta caatttaaat
     1321 gtaacccatc catccattca tccatccatc catccactca ttcatattga agttccagtc
     1381 tattgctaac acggaatttt aatttttctt tacaggagag ggtgaaggtg atgcaacata
     1441 cggaaaactt acccttaaat ttatttgcac tactggaaaa ctacctgttc cctggccaac
     1501 acttgtaact actttctgtt atggtgttca atgcttctcg agatacccag ataatatgaa
     1561 acggcatgac tttttcaaga gtgctatgcc cgaaggttat gtacaggaaa gaactatatt
     1621 tttcaaagat gacgggaact acaagacacg tgctgaagtc aagtttgaag gtgataccct
     1681 tgttaataga atcgagttaa aaggcattga ttttaaagaa gatggaaaca ttcttggaca
     1741 caaattggaa tacaactata acgcacacaa tgtatacatt atggcagaca aacaaaagaa
     1801 tggaatcaaa gttaacttca aaattagaca caacattgaa gatggaagcg ttcaactagc
     1861 agacaattat caacaaaata ctccaattgg cgatggccct gtccttttac cagtaagtta
     1921 aaacttgatt cagtgacctc ttttatctat tctaacatac tgccatccat ccattcatcc
     1981 atccatccat ccactcattc atgttaaaaa tacagactat aatttaattt agcacgtcca
     2041 atggaaagct ggccaacatg gtgaaacctt gtctctacta aaaatatgaa aattagccag
     2101 gcatggtggc acattcctgt aatcccagga ggctgaggca ggagaataac ttgaacccag
     2161 gaggtggagg ttgcagcaag ccgagatcgt acaactgtgc tctagccttg gtgacagagc
     2221 gagactgtct taaaaaaaat gttgctctaa cattgtcaat ttaaaatatc tacttttgtt
     2281 tcgtaacagg acaacaatta cctgtctaca caatctgccc tttcgaaaga tcccaacgaa
     2341 aagagagaca acatggtcct tcttgagttt gtaacagctg ctgggattac acatggcatg
     2401 gatgaactat acaaatagat caataacact ggcggccgct cgagcatgca tctagagggc
     2461 cctattctat agtgtcacct aaatgctaga gctcgctgat cagcctcgac tgtgccttct
     2521 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     2581 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     2641 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     2701 agcaggcatg ctggggatgc ggtgggctct atgggttcac tcctcaggtg caggctgcct
     2761 atcagaaggt ggtggctggt gtggccaatg ccctggctca caaataccac tgagatcttt
     2821 ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct
     2881 aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg
     2941 gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt
     3001 tggcaacata tgcccatatg ctggctgcca tgaacaaagg ttggctataa agaggtcatc
     3061 agtatatgaa acagccccct gctgtccatt ccttattcca tagaaaagcc ttgacttgag
     3121 gttagatttt ttttatattt tgttttgtgt tatttttttc tttaacatcc ctaaaatttt
     3181 ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg
     3241 tccctcttct cttatggaga tccgcggaac ccctatttgt ttatttttct aaatacattc
     3301 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     3361 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     3421 ccttcctgtt tttgctcacc ccgaaacgct ggtgaaagta aaagatgctg aagatcagtt
     3481 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggcaagatcc ttgagagttt
     3541 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     3601 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     3661 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     3721 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     3781 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     3841 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     3901 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     3961 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     4021 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     4081 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     4141 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     4201 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     4261 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     4321 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     4381 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     4441 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     4501 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     4561 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     4621 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     4681 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     4741 cagcttggag cgaacgacct acaccgaact
//
