LOCUS       sequence-358815-        4690 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses GFP CDS-derived linear and circular RNA transcript. 
            Flanking introns are devoid of NOVA2 binding sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4690)
  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 4690)
  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-WT
FEATURES             Location/Qualifiers
     source          1..4690
                     /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     2275..2295
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     promoter        complement(2386..2404)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2387..2404)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2424..2441)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2430..2654
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2736..2755)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2801..2856
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2855..2874)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     promoter        3183..3287
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3288..3356,3357..4148)
                     /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(3506..3525)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4319..4690,1..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 gtaacattga agttccagtc tattgctaac acggaatttt aatttttctt tacaggagag
     1381 ggtgaaggtg atgcaacata cggaaaactt acccttaaat ttatttgcac tactggaaaa
     1441 ctacctgttc cctggccaac acttgtaact actttctgtt atggtgttca atgcttctcg
     1501 agatacccag ataatatgaa acggcatgac tttttcaaga gtgctatgcc cgaaggttat
     1561 gtacaggaaa gaactatatt tttcaaagat gacgggaact acaagacacg tgctgaagtc
     1621 aagtttgaag gtgataccct tgttaataga atcgagttaa aaggcattga ttttaaagaa
     1681 gatggaaaca ttcttggaca caaattggaa tacaactata acgcacacaa tgtatacatt
     1741 atggcagaca aacaaaagaa tggaatcaaa gttaacttca aaattagaca caacattgaa
     1801 gatggaagcg ttcaactagc agacaattat caacaaaata ctccaattgg cgatggccct
     1861 gtccttttac cagtaagtta aaacttgatt cagtgacctc ttttatctat tctaacatag
     1921 tggttaaaaa tacagactat aatttaattt agcacgtcca atggaaagct ggccaacatg
     1981 gtgaaacctt gtctctacta aaaatatgaa aattagccag gcatggtggc acattcctgt
     2041 aatcccagga ggctgaggca ggagaataac ttgaacccag gaggtggagg ttgcagcaag
     2101 ccgagatcgt acaactgtgc tctagccttg gtgacagagc gagactgtct taaaaaaaat
     2161 gttgctctaa cattgtcaat ttaaaatatc tacttttgtt tcgtaacagg acaacaatta
     2221 cctgtctaca caatctgccc tttcgaaaga tcccaacgaa aagagagaca acatggtcct
     2281 tcttgagttt gtaacagctg ctgggattac acatggcatg gatgaactat acaaatagat
     2341 caataacact ggcggccgct cgagcatgca tctagagggc cctattctat agtgtcacct
     2401 aaatgctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     2461 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     2521 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     2581 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     2641 ggtgggctct atgggttcac tcctcaggtg caggctgcct atcagaaggt ggtggctggt
     2701 gtggccaatg ccctggctca caaataccac tgagatcttt ttccctctgc caaaaattat
     2761 ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc
     2821 attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca
     2881 aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata tgcccatatg
     2941 ctggctgcca tgaacaaagg ttggctataa agaggtcatc agtatatgaa acagccccct
     3001 gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt ttttatattt
     3061 tgttttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt tttactagcc
     3121 agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct cttatggaga
     3181 tccgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3241 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     3301 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     3361 ccgaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     3421 tcgaactgga tctcaacagc ggcaagatcc ttgagagttt tcgccccgaa gaacgttttc
     3481 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     3541 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     3601 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     3661 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     3721 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     3781 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     3841 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     3901 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     3961 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     4021 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     4081 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     4141 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     4201 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     4261 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     4321 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     4381 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     4441 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     4501 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     4561 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     4621 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     4681 acaccgaact
//
