LOCUS       sequence-357447-        3354 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ms2 incorporated pegRNA-1.1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3354)
  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 3354)
  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: MS2-pegRNA
FEATURES             Location/Qualifiers
     source          1..3354
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      358..946
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     847..866
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1100..1117
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1234..1255
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1270..1275,1276..1293,1294..1300)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1308..1324
                     /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     1313..1335
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1332..1348
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1332..1348
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1367..1387
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1369..1387
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1434..1682
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1434..1454
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1605..1624
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        1721..1739
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     primer_bind     complement(1867..1883)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1868..1884)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(1908..1927)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1909..1927)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1937..1954)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1937..1953)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1946..1968)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2095..2550
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2182..2201)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2392..2413
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2576..2680
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2681..2749,2750..3354,1..187)
                     /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     complement(2899..2918)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
       61 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
      121 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
      181 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
      241 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
      301 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
      361 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
      421 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
      481 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
      541 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
      601 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
      661 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
      721 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
      781 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
      841 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
      901 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
      961 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     1021 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     1081 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     1141 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     1201 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
     1261 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     1321 acaatttcac acaggaaaca gctatgacca tgattacgcc aagcgcgcaa ttaaccctca
     1381 ctaaagggaa caaaagctgg agctccaccg cggtggcggc cgcccccttc accgagggcc
     1441 tatttcccat gattccttca tatttgcata tacgatacaa ggctgttaga gagataattg
     1501 gaattaattt gactgtaaac acaaagatat tagtacaaaa tacgtgacgt agaaagtaat
     1561 aatttcttgg gtagtttgca gttttaaaat tatgttttaa aatggactat catatgctta
     1621 ccgtaacttg aaagtatttc gatttcttgg ctttatatat cttgtggaaa ggacgaaaca
     1681 ccgcaccttc agcttggcgg tctgttttag agctaggcca acatgaggat cacccatgtc
     1741 tgcagggcct agcaagttaa aataaggcta gtccgttatc acgccgaaag gcgggcaccg
     1801 agtcggtgct acgagggcac tcaaaccgcc aagctgaagt ttttttgaat tcgatatcaa
     1861 gcttatcgat accgtcgacc tcgagggggg gcccggtacc caattcgccc tatagtgagt
     1921 cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     1981 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     2041 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc
     2101 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     2161 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     2221 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     2281 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     2341 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     2401 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     2461 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     2521 attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg
     2581 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     2641 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     2701 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     2761 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     2821 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     2881 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     2941 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     3001 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     3061 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     3121 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     3181 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     3241 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     3301 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggc
//
