LOCUS       sequence-371448-        2486 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Part 3-4-5 plasmid to introduce 3xFLAG-NanoLuc into circRNAs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2486)
  AUTHORS   Chen R, Wang SK, Belk JA, Amaya L, Li Z, Cardenas A, Abe BT, Chen 
            CK, Wender PA, Chang HY
  TITLE     Engineering circular RNA for enhanced protein production.
  JOURNAL   Nat Biotechnol. 2022 Jul 18. pii: 10.1038/s41587-022-01393-0. doi: 
            10.1038/s41587-022-01393-0.
  PUBMED    35851375
REFERENCE   2  (bases 1 to 2486)
  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: "Nat 
            Biotechnol. 2022 Jul 18. pii: 10.1038/s41587-022-01393-0. doi: 
            10.1038/s41587-022-01393-0."
COMMENT     Sequence Label: 3xFLAG-NanoLuc
FEATURES             Location/Qualifiers
     source          1..2486
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(15..119)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      222..810
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     711..730
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             936..1001
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1017..1529
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     CDS             complement(join(1640..2431,2432..2486,1..14))
                     /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     2263..2282
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
       61 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
      121 acatttcccc gaaaagtgcc acctgtcatg accaaaatcc cttaacgtga gttttcgttc
      181 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
      241 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
      301 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
      361 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
      421 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
      481 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
      541 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
      601 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
      661 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
      721 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
      781 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
      841 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
      901 gataaccgtg cggccgctcg gtctcaatgg gatccgacta taaggaccac gacggagact
      961 acaaggatca tgatattgat tacaaggacg acgatgacaa gggaggagga ggaagcgtct
     1021 tcacactcga agatttcgtt ggggactggc gacagacagc cggctacaac ctggaccaag
     1081 tccttgaaca gggaggtgtg tccagtttgt ttcagaatct cggggtgtcc gtaactccga
     1141 tccaaaggat tgtcctgagc ggtgaaaatg ggctgaagat cgacatccat gtcatcatcc
     1201 cgtatgaagg tctgagcggc gaccaaatgg gccagatcga aaaaattttt aaggtggtgt
     1261 accctgtgga tgatcatcac tttaaggtga tcctgcacta tggcacactg gtaatcgacg
     1321 gggttacgcc gaacatgatc gactatttcg gacggccgta tgaaggcatc gccgtgttcg
     1381 acggcaaaaa gatcactgta acagggaccc tgtggaacgg caacaaaatt atcgacgagc
     1441 gcctgatcaa ccccgacggc tccctgctgt tccgagtaac catcaacgga gtgaccggct
     1501 ggcggctgtg cgaacgcatt ctggcgtagt tgagaccgcg gccgcgatta tcaaaaagga
     1561 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     1621 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     1681 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     1741 agggcttacc atctggcccc agtgctgcaa tgataccgcg ggacccacgc tcaccggctc
     1801 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     1861 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     1921 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     1981 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     2041 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     2101 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     2161 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     2221 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     2281 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     2341 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     2401 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     2461 aaaagggaat aagggcgaca cggaaa
//
