LOCUS       sequence-254691-        6242 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  circular Pepper RNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6242)
  AUTHORS   Wu J, Zaccara S, Khuperkar D, Kim H, Tanenbaum ME, Jaffrey SR
  TITLE     Live imaging of mRNA using RNA-stabilized fluorogenic proteins.
  JOURNAL   Nat Methods. 2019 Sep;16(9):862-865. doi: 10.1038/s41592-019-0531-7.
            Epub 2019 Aug 30.
  PUBMED    31471614
REFERENCE   2  (bases 1 to 6242)
  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: 
            "10.1038/s41592-019-0531-7"; journalName: "Nat Methods"; date: 
            "2019-09"; volume: "16"; issue: "9"; pages: "862-865"
COMMENT     Sequence Label: pAV-U6+27-Tornado-F30-Pepper(TAR Variant-2)
FEATURES             Location/Qualifiers
     source          1..6242
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..330
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      181..316
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             687..1481
                     /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(741..760)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1351..1370
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     intron          1901..1966
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             2096..2116
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2541..2675
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2944..3532)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3024..3043)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3703..4494,4495..4563))
                     /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     4326..4345
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4564..4668)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4695..5150
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4782..4801)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4992..5013
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     repeat_region   5244..5373
                     /label=AAV2 ITR
     polyA_signal    5422..5543
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(5962..6210)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(6020..6039)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(6190..6210)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(join(6238..6242,1..16))
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
       61 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
      121 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
      181 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
      241 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
      301 ggcttttttg gaggcctagg cttttgcaaa aagcttcacg ctgccgcaag cactcagggc
      361 gcaagggctg ctaaaggaag cggaacacgt agaaagccag tccgcagaaa cggtgctgac
      421 cccggatgaa tgtcagctac tgggctatct ggacaaggga aaacgcaagc gcaaagagaa
      481 agcaggtagc ttgcagtggg cttacatggc gatagctaga ctgggcggtt ttatggacag
      541 caagcgaacc ggaattgcca gctggggcgc cctctggtaa ggttgggaag ccctgcaaag
      601 taaactggat ggctttcttg ccgccaagga tctgatggcg caggggatca agatctgatc
      661 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
      721 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
      781 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
      841 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
      901 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
      961 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     1021 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     1081 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     1141 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     1201 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     1261 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     1321 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     1381 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     1441 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     1501 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     1561 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     1621 cggggatctc atgctggagt tcttcgccca ccccgggctc gatcccctcg cgagttggtt
     1681 cagctgctgc ctgaggctgg acgacctcgc ggagttctac cggcagtgca aatccgtcgg
     1741 catccaggaa accagcagcg gctatccgcg catccatgcc cccgaactgc aggagtgggg
     1801 aggcacgatg gccgctttgg tccggatctt tgtgaaggaa ccttacttct gtggtgtgac
     1861 ataattggac aaactaccta cagagattta aagctctaag gtaaatataa aatttttaag
     1921 tgtataatgt gttaaactac tgattctaat tgtttgtgta ttttagattc caacctatgg
     1981 aactgatgaa tgggagcagt ggtggaatgc ctttaatgag gaaaacctgt tttgctcaga
     2041 agaaatgcca tctagtgatg atgaggctac tgctgactct caacattcta ctcctccaaa
     2101 aaagaagaga aaggtagaag accccaagga ctttccttca gaattgctaa gttttttgag
     2161 tcatgctgtg tttagtaata gaactcttgc ttgctttgct atttacacca caaaggaaaa
     2221 agctgcactg ctatacaaga aaattatgga aaaatattct gtaaccttta taagtaggca
     2281 taacagttat aatcataaca tactgttttt tcttactcca cacaggcata gagtgtctgc
     2341 tattaataac tatgctcaaa aattgtgtac ctttagcttt ttaatttgta aaggggttaa
     2401 taaggaatat ttgatgtata gtgccttgac tagagatcat aatcagccat accacatttg
     2461 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     2521 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     2581 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     2641 ccaaactcat caatgtatct tatcatgtct ggatcgatcc aggacctccc taaccctatg
     2701 acgtaattca cgtcacgact ccacccctcc aggaacccct agtgatggag ttggccactc
     2761 cctctctgcg cgctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     2821 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     2881 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     2941 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3001 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3061 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3121 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3181 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3241 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3301 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3361 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     3421 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3481 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     3541 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3601 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     3661 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     3721 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     3781 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     3841 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     3901 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     3961 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     4021 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4081 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     4141 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     4201 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     4261 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     4321 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     4381 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     4441 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     4501 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     4561 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     4621 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     4681 aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     4741 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     4801 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     4861 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     4921 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     4981 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     5041 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     5101 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt ccattcgcca
     5161 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     5221 cagatctgca gatctgtccc tctctgcgcg ctcgctcgct cactgaggcc gcccgggcaa
     5281 agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag
     5341 agggagtggc caactccatc actaggggtt ccttgtagtt aatgattaac ccgccatgct
     5401 acttatctac gtaagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     5461 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     5521 ttataagctg caataaacaa gttactagtc ctcggctcga gatcgatgcg gtcgccatat
     5581 ggatatcggt aaccgacgtc aagcttgcat gcctgcaggt cctagtaaaa agcggaccga
     5641 agtccgctct agagcgtgga ctgtaccctc cacttttatc cgggcttggg accggcattg
     5701 gcagtgttct acagtccacg ccgaccgcgg ccgcttgcca tgaatgatcc ggtacctcgg
     5761 gctcggagct aatgagctca acgagccttt gctagcggat catcagagta tgtgggaaac
     5821 gtggcaacgc gtcccacata cacatggcaa gcggccgcac tcggcatggt taggcggttt
     5881 cccgccccct cccattttat ccgggcttgg gaccggcgag tgcggcccgt cgactagtat
     5941 atgtgctgcc gaagcgagca cggtgtttcg tcctttccac aagatatata aagccaagaa
     6001 atcgaaatac tttcaagtta cggtaagcat atgatagtcc attttaaaac ataattttaa
     6061 aactgcaaac tacccaagaa attattactt tctacgtcac gtattttgta ctaatatctt
     6121 tgtgtttaca gtcaaattaa ttctaattat ctctctaaca gccttgtatc gtatatgcaa
     6181 atatgaagga atcatgggaa ataggccctc ttcctgcccg accttggatc cggctgtgga
     6241 at
//
