LOCUS       sequence-357444-        4807 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E.r. maturase RT .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4807)
  AUTHORS   Liu B, Dong X, Cheng H, Zheng C, Chen Z, Rodriguez TC, Liang SQ, Xue
            W, Sontheimer EJ
  TITLE     A split prime editor with untethered reverse transcriptase and 
            circular RNA template.
  JOURNAL   Nat Biotechnol. 2022 Apr 4. pii: 10.1038/s41587-022-01255-9. doi: 
            10.1038/s41587-022-01255-9.
  PUBMED    35379962
REFERENCE   2  (bases 1 to 4807)
  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 Apr 4. pii: 10.1038/s41587-022-01255-9. doi: 
            10.1038/s41587-022-01255-9."
COMMENT     Sequence Label: maturase-RT
FEATURES             Location/Qualifiers
     source          1..4807
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(124..228)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(307..326)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        498..877
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        878..1081
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1032..1052
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1123..1132
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1132..1197
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1204..1224
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2539..2559
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2568..2585
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2608..2625)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2614..2838
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2909..2925)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2909..2925)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2922..2944)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2933..2949
                     /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)."
     promoter        complement(join(2957..2963,2964..2981,2982..2987))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3002..3023
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3140..3157)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3311..3899)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3391..3410)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4070..4807,1..54,55..123))
                     /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     4693..4712
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
       61 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
      121 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
      181 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
      241 aaaagtgcca cctgacgtcg acggatcggg agatcgatct cccgatcccc tagggtctac
      301 tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt
      361 gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac
      421 cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg
      481 ggccagatat acgcgttgac attgattatt gactagttat taatagtaat caattacggg
      541 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      601 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      661 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      721 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
      781 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
      841 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
      901 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      961 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
     1021 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
     1081 tggtttagtg aaccgtcaga tccgctagag atccgcggcc gcgccaccat ggactataag
     1141 gaccacgacg gagactacaa ggatcatgat attgattaca aagacgatga cgataagatg
     1201 gccccaaaga agaagcggaa ggtcggtatc cacggagtcc cagcagccga taccagcaat
     1261 ctgatggaac aaattctgag cagcgataat ctgaatcgtg catatctgca ggttgtgcgt
     1321 aataaaggtg ccgaaggtgt tgatggtatg aaatataccg aactgaaaga acacctggcc
     1381 aaaaatggtg aaaccattaa aggtcagctg cgtaccgccg cctacaaacc gcagcctgca
     1441 cgtcgtgttg aaattccgaa accggatggt ggtgttcgta atctgggtgt tccgaccgtt
     1501 accgatcgtt ttattcagca ggcaattgca caggttctga ccccgattta tgaagaacag
     1561 tttcatgatc acagctatgg ctttcgtccg aatcgttgtg cacagcaggc cattctgacc
     1621 gcactgaata ttatgaatga tggcaatgat tggatcgtgg atatcgacct ggaaaaattc
     1681 tttgataccg tgaaccacga taaactgatg accctgattg gtcgtacaat cgccgatggt
     1741 gatgtgatta gcatcgtgcg taaatatctg gttagcggca ttatgatcga tgatgaatat
     1801 gaggatagca ttgttggtac accgcagggt ggtaatctga gtccgctgct ggccaatatt
     1861 atgctgaatg aactggacaa agagatggaa gccgccggtc tgaattttgt tcgttatgcc
     1921 gatgattgca ttattatggt tggtagcgaa atgagcgcaa atcgtgttat gcgtaatatt
     1981 agccgcttca tcgaagaaaa actgggcctg aaagttaaca tgaccaaaag caaagttgat
     2041 cgtccgagtg gtctgaaata cctgggtttt ggtttctatt ttgatccgcg tgcacaccag
     2101 tttaaagcaa aaccgcatgc aaaaagcgtg gccaaattca aaaaacgcat gaaagaactg
     2161 acctgtcgta gctggggtgt tagcaatagc tataaagtgg aaaaactgaa ccagctgatt
     2221 cgcggttgga tcaactattt caaaattggc agcatgaaaa ccctgtgcaa agaactggat
     2281 agccgtattc gttatcgtct gcgtatgtgt atttggaaac agtggaaaac accgcagaat
     2341 caagaaaaaa atctggtgaa actgggcatt gatcgtaata ccgcacgtcg cgttgcatat
     2401 accggtaaac gtattgcata tgtgtgcaat aaaggcgcag ttaatgttgc cattagcaat
     2461 aaacgtctgg caagctttgg tctgattagt atgctggatt actacatcga aaaatgcgtg
     2521 acctgcagcg aattcgagcc caagaagaag aggaaagtct aaccggtcat catcaccatc
     2581 accattgagt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     2641 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     2701 cctaataaaa tgagaaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     2761 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     2821 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctcg ataccgtcga
     2881 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     2941 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tagggtgcct
     3001 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     3061 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     3121 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     3181 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     3241 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     3301 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     3361 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     3421 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     3481 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     3541 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     3601 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     3661 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     3721 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     3781 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     3841 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     3901 aagatccttt gatcttttct acggggtctg acactcagtg gaacgaaaac tcacgttaag
     3961 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     4021 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     4081 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     4141 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     4201 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     4261 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     4321 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     4381 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     4441 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     4501 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     4561 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     4621 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     4681 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     4741 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     4801 aacccac
//
