LOCUS       sequence-374478-        7218 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template plasmid to transcribe circular RNA encoding mNeonGreen.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7218)
  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 7218)
  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: circRNA-synIRES-R25-mNeonGreen
FEATURES             Location/Qualifiers
     source          1..7218
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        66..369
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          647..1663
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1587..1609
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1671..1690
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     1733..1752
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1733..1751
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      3920..3967
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3981..4205
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    4248..4264
                     /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)."
     primer_bind     4253..4275
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4272..4288
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4272..4288
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    4337..4458
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4348..4367)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4402..4421
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(4618..5217)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4701..4721)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     5198..5217
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(5228..5556)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5242..5377
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5296..5315)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             complement(5590..6405)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(5590..6405)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     6313..6332
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6397..6416
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6630..7218
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7119..7138
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
       61 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      121 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      181 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      241 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      301 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      361 attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc
      421 ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg
      481 gggggggggg ggcgcgccrg gggggggggg gggggggggg gggggggggg gggsgrggsg
      541 grgrggkgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag
      601 gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg
      661 ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact
      721 gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta
      781 gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct
      841 ccgggagggc cctttgtgcg gggggagcgg ctcggggggt gcgtgcgtgt gtgtgtgcgt
      901 ggggagcgcc gcgtgcggct ccgcgctgcc cggcggctgt gagcgctgcg ggcgcggcgc
      961 ggggctttgt gcgctccgca gtgtgcgcga ggggagcgcg gccgggggcg gtgccccgcg
     1021 gtgcgggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg tgggggggtg
     1081 agcagggggt gtgggcgcgt cggtcgggct gcaacccccc ctgcaccccc ctccccgagt
     1141 tgctgagcac ggcccggctt cgggtgcggg gctccgtacg gggcgtggcg cggggctcgc
     1201 cgtgccgggc ggggggtggc ggcaggtggg ggtgccgggc ggggcggggc cgcctcgggc
     1261 cggggagggc tcgggggagg ggcgcggcgg cccccggagc gccggcggct gtcgaggcgc
     1321 ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg gacttccttt
     1381 gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct ctagcgggcg
     1441 cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc ttcgtgcgtc
     1501 gccgcgccgc cgtccccttc tccctctcca gcctcggggc tgtccgcggg gggacggctg
     1561 ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg gcggctctag
     1621 agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg gcaacgtgct
     1681 ggttattgtg ctgtctcatc attttggcaa agcggccgcg gccagtgaat tgtaatacga
     1741 ctcactatag gggttaacaa aaaaaaaaaa aaaaaaaaaa aatcaccgac cgatctatct
     1801 gaaccggtgg gaattctaga gaaaatttcg tctggattag ttacttatcg tgtaaaatct
     1861 gataaatgga attggttcta cataaatgcc taacgactat ccctttgggg agtagggtca
     1921 agtgactcga aacgatagac aacttgcttt aacaagttgg agatatagtc tgctctgcat
     1981 ggtgacatgc agctggatat aattccgggg taagattaac gaccttatct gaacataacg
     2041 ctaccgttta atattgcgtc atattcggcg accatttgtg tggtaaaaaa aaaaaaccaa
     2101 aaaaaaaaaa caaaaaaaaa aaataattga ctaagatatc ttaaaacagc ggatgggtac
     2161 cccaccatcc gacccactgg gtgtagtact ctggtacttc gtacctttgt acgcctgttc
     2221 ttcccattgt acccttcctg aacttccaac ccaagtaacg ttagaagctc aacatttagt
     2281 acaacaggaa gcaccacatc cagtggtgtt tagtacaagc acttctgttt ccccggagcg
     2341 aggtataggc tgtacccact gccaaaaacc tttaaccgtt atccgccaac caactacgta
     2401 aaagctagta gtattatgtt tttaactagg cgttcgatca ggtggatttc ccctccacta
     2461 gtttggtcga tgaggctagg aattccccac gggtgaccgt gtcctagcct gcgtggcggc
     2521 caacccagcc cactcactat ttgttttcgc gcccagttgc aaaaagtgtc ggggctggga
     2581 cgccttttta tagacatggt gtgaagactc gcatgtgctt ggttgtgatt cctccggccc
     2641 ctgaatgcgg ctaaccttaa ccctggagcc ttgtgtcaca aaccagtgat gataaggtcg
     2701 taatgagcaa ttccgggacg ggaccgacta ctttgggtgt ccgtgtttct tatttttctt
     2761 attattgtct tatggtcaca gcatatatat aacatatact gtgatcatgg gatccgtgtc
     2821 taaaggagag gaagataaca tggcgagcct cccagccact catgaactcc acatattcgg
     2881 aagcatcaac ggggtggact ttgacatggt aggtcagggg accggcaatc ccaacgacgg
     2941 ttacgaggaa ttgaatctca agtccactaa gggcgacctc cagttttctc cgtggattct
     3001 cgtcccacat atcgggtatg gtttccacca atacttgcct tacccggatg gaatgtcacc
     3061 gttccaggct gccatggttg atggctctgg ttatcaggta cacagaacga tgcaatttga
     3121 ggacggagcg tcattgactg taaactacag atacacatac gaaggctctc acataaaggg
     3181 cgaagcacaa gttaaaggca cgggattccc cgctgatggt cctgtcatga caaattctct
     3241 taccgcagcc gactggtgtc gatccaagaa aacgtatccc aatgacaaaa ctatcatatc
     3301 tacgttcaaa tggtcatata caacgggaaa cggtaagaga taccggagca ctgccagaac
     3361 cacctataca tttgccaaac caatggcggc taactacctg aagaaccagc ccatgtatgt
     3421 gttcaggaaa acggaactga aacactccaa gacagagctg aatttcaagg aatggcaaaa
     3481 agccttcacg gatgttatgg ggatggatga gctgtataaa tagtaacaat tggctggagc
     3541 ctcggtggcc atgcttcttg ccccttgggc ctccccccag cccctcctcc ccttcctgca
     3601 cccgtacccc cgtggtcttt gaataaagtc tgaaccacac aaatggtcgc cgactcagta
     3661 gatgttttct tgggttaatt gaggcctgag tataaggtga cttatacttg taatctatct
     3721 aaacggggaa cctctctagt agacaatccc gtgctaaatt gtaggactgc cctttaataa
     3781 atacttctat atttaaagag gtatttatga aaagcggaat ttatcagatt aaaaatactt
     3841 tctctagagt cgacctgcag acgcgttcag atagatcggt cggtgaaaaa aaaaaaaaaa
     3901 aaaaaaaaaa agcggccgcc tagcataacc ccttggggcc tctaaacggg tcttgagggg
     3961 ttttttggct agcggccggc ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     4021 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     4081 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     4141 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     4201 tatggcggcg cgccacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     4261 acaatttcac acaggaaaca gctatgacca tgattacgcc aagctctagc tagaggtcga
     4321 cggtatacag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     4381 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     4441 agctgcaata aacaagttgg ggtgggcgaa gaactccagc atgagatccc cgcgctggag
     4501 gatcatccag ccggcgtccc ggaaaacgat tccgaagccc aacctttcat agaaggcggc
     4561 ggtggaatcg aaatctcgtg atggcaggtt gggcgtcgct tggtcggtca tttcgaatca
     4621 ggcaccgggc ttgcgggtca tgcaccaggt gcgcggtcct tcgggcacct cgacgtcggc
     4681 ggtgacggtg aagccgagcc gctcgtagaa ggggaggttg cggggcgcgg aggtttccag
     4741 gaaggcgggc accccggcgc gctcggccgc ctccactccg gggagcacga cggcgctgcc
     4801 cagacccttg ccctggtggt cgggcgacac gccgacggtg gccaggaacc acgcgggctc
     4861 cttgggccgg tgcggcgcca ggaggccttc catctgttgc tgcgcggcca gccgggaacc
     4921 gctcaactcg gccatgcgcg ggccgatctc ggcgaacacc gcccccgctt cgacgctctc
     4981 cggcgtggtc cagaccgcca ccgcggcgcc gtcgtccgcg acccacacct tgccgatgtc
     5041 gagcccgacg cgcgtgagga agagttcttg cagctcggtg acccgctcga tgtggcggtc
     5101 cggatcgacg gtgtggcgcg tggcggggta gtcggcgaac gcggcggcga gggtgcgtac
     5161 ggccctgggg acgtcgtcgc gggtggcgag gcgcaccgtg ggcttgtact cggtcatgcg
     5221 ggagcttttt gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct
     5281 cagaggcaga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg
     5341 gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta
     5401 tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg
     5461 actttccaca cctggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg
     5521 gggagcctgg ggactttcca caccctaact gacacagagc tcgtgttaca accaattaac
     5581 caattctgat tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg
     5641 attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag
     5701 gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc
     5761 aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg
     5821 agtgacgact gaatccggtg agaatggcaa aagcttatgc atttctttcc agacttgttc
     5881 aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat
     5941 tcgtgattgc gcctgagcga ggcgaaatac gcgatcgctg ttaaaaggac aattacaaac
     6001 aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga
     6061 atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag tggtgagtaa
     6121 ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt
     6181 cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg
     6241 tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga
     6301 ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt
     6361 taatcgcggc ctggagcaag acgtttcccg ttgaatatgg ctcataacac cccttgtatt
     6421 actgtttatg taagcagaca gttttattgt tcatgatgat atatttttat cttgtgcaat
     6481 gtaacatcag agattttgag acacaacgtg gctttgttga ataaatcgaa cttttgctga
     6541 gttgaaggat cagtcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     6601 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     6661 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     6721 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     6781 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     6841 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     6901 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     6961 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     7021 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     7081 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     7141 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     7201 gggcggagcc tatggaaa
//
