LOCUS       sequence-424997-        6185 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  encodes T5 exonuclease with a PP7 coat protein fused to the N 
            terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6185)
  AUTHORS   Truong DJ, Geilenkeuser J, Wendel SV, Wilming JCH, Armbrust N, 
            Binder EMH, Santl TH, Siebenhaar A, Gruber C, Phlairaharn T, Zivanic
            M, Westmeyer GG
  TITLE     Exonuclease-enhanced prime editors.
  JOURNAL   Nat Methods. 2024 Feb 1. doi: 10.1038/s41592-023-02162-w.
  PUBMED    38302659
REFERENCE   2  (bases 1 to 6185)
  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 
            Methods. 2024 Feb 1. doi: 10.1038/s41592-023-02162-w."
COMMENT     Sequence Label: pCAG_PCP-T5Exo
FEATURES             Location/Qualifiers
     source          1..6185
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        23..402
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        404..680
                     /label=chicken beta-actin promoter
     intron          681..1697
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1621..1643
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1710..1729
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1710..1728
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2117..2137
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2153..2179
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             2177..2200
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3091..3107)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3121..3138)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3127..3351
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3423..3440)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3423..3439)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3432..3454)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3580..4035
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3667..3686)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3877..3898
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4062..4166
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4167..4235,4236..5027)
                     /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(4385..4404)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5198..5786
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5687..5706
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5940..5957
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6074..6095
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6110..6115,6116..6133,6134..6140)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6148..6164
                     /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     6153..6175
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(6172..6185,1..3)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(6172..6185,1..3)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gaccatgagg cgcgccggat tcgacattga ttattgacta gttattaata gtaatcaatt
       61 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
      121 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
      181 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
      241 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
      301 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
      361 acttggcagt acatctacgt attagtcatc gctattacca tggtcgaggt gagccccacg
      421 ttctgcttca ctctccccat ctcccccccc tccccacccc caattttgta tttatttatt
      481 ttttaattat tttgtgcagc gatgggggcg gggggggggg ggggcgcgcg ccaggcgggg
      541 cggggcgggg cgaggggcgg ggcggggcga ggcggagagg tgcggcggca gccaatcaga
      601 gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa
      661 agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg
      721 ccgccgcctc gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg
      781 ggcgggacgg cccttctcct ccgggctgta attagcgctt ggtttaatga cggcttgttt
      841 cttttctgtg gctgcgtgaa agccttgagg ggctccggga gggccctttg tgcgggggga
      901 gcggctcggg gggtgcgtgc gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc
      961 tgcccggcgg ctgtgagcgc tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc
     1021 gcgaggggag cgcggccggg ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca
     1081 aaggctgcgt gcggggtgtg tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg
     1141 ggctgcaacc ccccctgcac ccccctcccc gagttgctga gcacggcccg gcttcgggtg
     1201 cggggctccg tacggggcgt ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg
     1261 tgggggtgcc gggcggggcg gggccgcctc gggccgggga gggctcgggg gaggggcgcg
     1321 gcggcccccg gagcgccggc ggctgtcgag gcgcggcgag ccgcagccat tgccttttat
     1381 ggtaatcgtg cgagagggcg cagggacttc ctttgtccca aatctgtgcg gagccgaaat
     1441 ctgggaggcg ccgccgcacc ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca
     1501 ggaaggaaat gggcggggag ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc
     1561 tccagcctcg gggctgtccg cggggggacg gctgccttcg ggggggacgg ggcagggcgg
     1621 ggttcggctt ctggcgtgtg accggcggct ctagagcctc tgctaaccat gttcatgcct
     1681 tcttcttttt cctacagatc cttaattaat aatacgactc actatagggg ccgccaccat
     1741 gtctaagaca atcgtgctgt ctgtgggcga agccaccaga acactgaccg agatccagag
     1801 cacagccgac agacagatct tcgaggaaaa agtgggccct ctcgtgggca gactgagact
     1861 gacagcttcc ctgagacaga acggcgccaa gaccgcctac agagtgaacc tgaagctgga
     1921 ccaggccgac gtggtggatt ctggccttcc taaagtgcgg tacacccaag tgtggtccca
     1981 cgacgtgacc atcgtggcca attctaccga ggccagcaga aagagcctgt acgacctgac
     2041 caagagcctg gtggccacaa gccaggtgga agatctggtc gtgaacctgg tgcctctcgg
     2101 acgtggatcc acacctccta agaagaagcg gaaggtcgag gacggcgaag gacctgccgc
     2161 caagagagtg aaactggact acaaggacga cgacgacaag agcaagagct ggggcaagtt
     2221 catcgaagag gaagaggccg aggtggcttc ccggagaaat ttgatgatcg tggacggcac
     2281 caacctgggc ttcagattca agcacaacaa cagcaagaag cccttcgcca gcagctacgt
     2341 gtccaccatt cagagcctcg ccaagagcta cagcgcccgg acaacaattg tgctgggcga
     2401 caagggcaag tccgtgttca gactggaaca tctgcccgag tacaaaggca accgggacga
     2461 gaagtacgcc cagagaaccg aggaagagaa ggccctggac gagcagttct tcgagtacct
     2521 gaaggacgcc ttcgagctgt gcaagaccac ctttcctacc ttcaccatca gaggcgtcga
     2581 ggccgatgat atggccgcct acatcgtgaa gctgatcggc cacctgtacg atcacgtgtg
     2641 gctgatctcc accgacggcg attgggatac cctgctgacc gacaaggtgt cccggttcag
     2701 ctttaccacc agaagagagt accacctccg ggatatgtac gagcaccaca acgtggacga
     2761 cgtggaacag ttcatcagcc tgaaggccat catgggcgac ctgggcgata atatcagggg
     2821 cgttgaaggc atcggagcca agcggggcta caacatcatc cgcgagttcg gcaacgtgct
     2881 ggacatcatc gatcagctgc ctctgcctgg caagcagaag tacatccaga acctgaacgc
     2941 cagcgaggaa ctgctgttca gaaacctgat cctggtggac ctgcctacct actgcgtgga
     3001 cgctattgcc gccgttggac aggatgtgct ggataagttc accaaggaca tcctcgaaat
     3061 cgccgagcag tgaacgcgta aatgattgca gatccactag ttctagagct cgctgatcag
     3121 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     3181 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     3241 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     3301 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     3361 cggaaagaac cagctggggc tcgagatcca ctagttctag cctcgaggct agagcggccg
     3421 ccactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     3481 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     3541 cgcccttccc aacagttgcg cagcctgaat ggcgaatgga cgcgccctgt agcggcgcat
     3601 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     3661 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3721 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     3781 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     3841 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     3901 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     3961 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     4021 taacgcttac aatttcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     4081 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     4141 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     4201 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     4261 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     4321 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     4381 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     4441 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     4501 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     4561 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     4621 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     4681 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     4741 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     4801 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     4861 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     4921 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     4981 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     5041 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     5101 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     5161 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     5221 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     5281 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     5341 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     5401 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     5461 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     5521 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     5581 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     5641 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     5701 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     5761 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     5821 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     5881 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     5941 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     6001 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     6061 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
     6121 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
     6181 gctat
//
