LOCUS       sequence-373215-        4999 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Permuted td intron with evolved initiation sequence for the 
            expression of circular mRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4999)
  AUTHORS   Lee SO, Xie Q, Fried SD
  TITLE     Optimized Loopable Translation as a Platform for the Synthesis of 
            Repetitive Proteins.
  JOURNAL   ACS Cent Sci. 2021 Oct 27;7(10):1736-1750. doi: 
            10.1021/acscentsci.1c00574. Epub 2021 Sep 24.
  PUBMED    34729417
REFERENCE   2  (bases 1 to 4999)
  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: 
            "10.1021/acscentsci.1c00574"; journalName: "ACS Cent Sci"; date: 
            "2021-10-27- 27"; volume: "7"; issue: "10"; pages: "1736-1750"
COMMENT     Sequence Label: pBAD-tdTEVDB_C-15A/G4C
FEATURES             Location/Qualifiers
     source          1..4999
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             6..23
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             24..44
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     terminator      388..474
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(681..698)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(815..1360)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(895..914)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(1472..1477,1478..1494,1495..1500)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             1548..2738
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     complement(1900..1919)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2868..2890
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3046..3924)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        3951..4235
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     4158..4177
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
ORIGIN
        1 gcagccacca tcatcatcac catgaaaacc tgtattttca gggcaaagga gatatacata
       61 tgcctagcat gaatcacaaa ggtgaagaac tgtttaccgg cgttgtgccg attctggtgg
      121 aactggatgg tgatgtgaat ggccataaat ttagcgttcg tggcgaaggc gaaggtgatg
      181 cgaccaacgg taaactgacc ctgaaattta tttgcaccac cggttaattg aggcctgagt
      241 ataaggtgac ttatacttgt aatctatcta aacggggaac ctctctagta gacaatcccg
      301 tgctaaattg taggactatg catatcctta gcgaaagcta aggatttttt ttgtctcccc
      361 atgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg
      421 gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg
      481 ggagctgtcc ctcctgttca gctactgacg gggtggtgcg taacggcaaa agcaccgccg
      541 gacatcagcg ctagcggagt gtatactggc ttactatgtt ggcactgatg agggtgtcag
      601 tgaagtgctt catgtggcag gagaaaaaag gctgcaccgg tgcgtcagca gaatatgtga
      661 tacaggatat attccgcttc ctcgctcact gactcgctac gctcggtcgt tcgactgcgg
      721 cgagcggaaa tggcttacga acggggcgga gatttcctgg aagatgccag gaagatactt
      781 aacagggaag tgagagggcc gcggcaaagc cgtttttcca taggctccgc ccccctgaca
      841 agcatcacga aatctgacgc tcaaatcagt ggtggcgaaa cccgacagga ctataaagat
      901 accaggcgtt tccccctggc ggctccctcg tgcgctctcc tgttcctgcc tttcggttta
      961 ccggtgtcat tccgctgtta tggccgcgtt tgtctcattc cacgcctgac actcagttcc
     1021 gggtaggcag ttcgctccaa gctggactgt atgcacgaac cccccgttca gtccgaccgc
     1081 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg aaagacatgc aaaagcacca
     1141 ctggcagcag ccactggtaa ttgatttaga ggagttagtc ttgaagtcat gcgccggtta
     1201 aggctaaact gaaaggacaa gttttggtga ctgcgctcct ccaagccagt tacctcggtt
     1261 caaagagttg gtagctcaga gaaccttcga aaaaccgccc tgcaaggcgg ttttttcgtt
     1321 ttcagagcaa gagtttacgc gcagaccaaa acgatctcaa gaagatcatc ttattaatca
     1381 gataaaatat ttctagattt cagtgcaatt tatctcttca aatgtagcac ctgaagtcag
     1441 ccccatacga tataagttgt aattctcatg tttgacagct tatcatcgat aagctttaat
     1501 gcggtagttt atcacagtta aattgctaac gcagtcaggc accgtgtatg aaatctaaca
     1561 atgcgctcat cgtcatcctc ggcaccgtca ccctggatgc tgtaggcata ggcttggtta
     1621 tgccggtact gccgggcctc ttgcgggata tcgtccattc cgacagcatc gccagtcact
     1681 atggcgtgct gctagcgcta tatgcgttga tgcaatttct atgcgcaccc gttctcggag
     1741 cactgtccga ccgctttggc cgccgcccag tcctgctcgc ttcgctactt ggagccacta
     1801 tcgactacgc gatcatggcg accacacccg tcctgtggat cctctacgcc ggacgcatcg
     1861 tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg
     1921 atggggaaga tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc gtgggtatgg
     1981 tggcaggccc cgtggccggg ggactgttgg gcgccatctc cttgcatgca ccattccttg
     2041 cggcggcggt gctcaacggc ctcaacctac tactgggctg cttcctaatg caggagtcgc
     2101 ataagggaga gcgtcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt
     2161 gggcgcgggg catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg
     2221 taggacaggt gccggcagcg ctctgggtca ttttcggcga ggaccgcttt cgctggagcg
     2281 cgacgatgat cggcctgtcg cttgcggtat tcggaatctt gcacgccctc gctcaagcct
     2341 tcgtcactgg tcccgccacc aaacgtttcg gcgagaagca ggccattatc gccggcatgg
     2401 cggccgacgc gctgggctac gtcttgctgg cgttcgcgac gcgaggctgg atggccttcc
     2461 ccattatgat tcttctcgct tccggcggca tcgggatgcc cgcgttgcag gccatgctgt
     2521 ccaggcaggt agatgacgac catcagggac agcttcaagg atcgctcgcg gctcttacca
     2581 gcctaacttc gatcattgga ccgctgatcg tcacggcgat ttatgccgcc tcggcgagca
     2641 catggaacgg gttggcatgg attgtaggcg ccgccctata ccttgtctgc ctccccgcgt
     2701 tgcgtcgcgg tgcatggagc cgggccacct cgacctgaat ggaagccggc ggcacctcgc
     2761 taacggattc accactccaa gaattggagc caatcaattc ttgcggagaa ctgtgaatgc
     2821 gcaaaccaac ccttggcaga acatatccat cgcgtgcggc cgcgtctccg ggagctgcat
     2881 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagcagatca attcgcgcgc
     2941 gaaggcgaag cggcatgcat aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac
     3001 cctatgctac tccgtcaagc cgtcaattgt ctgattcgtt accaattatg acaacttgac
     3061 ggctacatca ttcacttttt cttcacaacc ggcacggaac tcgctcgggc tggccccggt
     3121 gcatttttta aatacccgcg agaaatagag ttgatcgtca aaaccaacat tgcgaccgac
     3181 ggtggcgata ggcatccggg tggtgctcaa aagcagcttc gcctggctga tacgttggtc
     3241 ctcgcgccag cttaagacgc taatccctaa ctgctggcgg aaaagatgtg acagacgcga
     3301 cggcgacaag caaacatgct gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg
     3361 atcgctgatg tactgacaag cctcgcgtac ccgattatcc atcggtggat ggagcgactc
     3421 gttaatcgct tccatgcgcc gcagtaacaa ttgctcaagc agatttatcg ccagcagctc
     3481 cgaatagcgc ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg
     3541 cggctggtgc gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg acggccagtt
     3601 aagccattca tgccagtagg cgcgcggacg aaagtaaacc cactggtgat accattcgcg
     3661 agcctccgga tgacgaccgt agtgatgaat ctctcctggc gggaacagca aaatatcacc
     3721 cggtcggcaa acaaattctc gtccctgatt tttcaccacc ccctgaccgc ggatggtgag
     3781 attgagaata taacctttca ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt
     3841 ggcctcaatc ggcgttaaac ccgccaccag atgggcatta aacgagtatc ccggcagaag
     3901 gggatcattt tgcgcttcag ccatactttt catactcccg ccattcagag aagaaaccaa
     3961 ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct tctcgctaac
     4021 caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca aagccatgac
     4081 aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg attatttgca
     4141 cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg atcctacctg
     4201 acgcttttta tcgcaactct ctactgtttc tccatacccg ttttttgggc tacgctgaat
     4261 tcgaattggt tctacataaa tgcctaacga ctatcccttt ggggagtagg gtcaagtgac
     4321 tcgaaacgat agacaacttg ctttaacaag ttggagatat agtctgctct gcatggtgac
     4381 atgcagctgg atataattcc ggggtaagat taacgacctt atctgaacat aatgctaaaa
     4441 ctgccggttc cgtggccgac cctggtgacc accttgacct atggcgttca gtgctttagc
     4501 cgctatccgg atcatatgaa acgccatgat ttctttaaaa gcgcgatgcc ggaaggctat
     4561 gtgcaggaac gtaccattag cttcaaagat gatggcacct ataaaacccg tgcggaagtt
     4621 aaatttgaag gcgataccct ggtgaaccgc attgaactga aaggtattga ttttaaagaa
     4681 gatggcaaca ttctgggtca taaactggaa tataatttca acagccataa cgtgtatatt
     4741 accgccgata aacagaaaaa tggcatcaaa gcgaacttta aaatccgtca caacgtggaa
     4801 gatggtagcg tgcagctggc ggatcattat cagcagaata ccccgattgg tgatggcccg
     4861 gtgctgctgc cggataatca ttatctgagc acccagagcg ttctgagcaa agatccgaat
     4921 gaaaaacgtg atcatatggt gctgctggaa tttgttaccg ccgcgggcat tacccacggt
     4981 atggatgaac tgtataaag
//
