LOCUS       sequence-183598-        3333 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAVS1-targeting plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3333)
  AUTHORS   Chen X, Janssen JM, Liu J, Maggio I, 't Jong AEJ, Mikkers HMM, 
            Goncalves MAFV
  TITLE     In trans paired nicking triggers seamless genome editing without 
            double-stranded DNA cutting.
  JOURNAL   Nat Commun. 2017 Sep 22;8(1):657. doi: 10.1038/s41467-017-00687-1.
  PUBMED    28939824
REFERENCE   2  (bases 1 to 3333)
  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 
            Commun."; date: "2017-09-22"; volume: "8(1):657. doi"; pages: " 
            10.1038/s41467-017-00687-1"
COMMENT     Sequence Label: AX44_pS.Donor.S1
FEATURES             Location/Qualifiers
     source          1..3333
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1112..1128)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1112..1128)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1125..1147)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1136..1152
                     /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(1160..1166,1167..1184,1185..1190))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1205..1226
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1343..1360)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1514..2102)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1594..1613)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2273..3064,3065..3133))
                     /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     2896..2915
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3134..3238)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3306..3324
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acctcgcgaa
      421 tgcatctaga tggccgcgga ttcgggtcac ctctcactcc tttcatttgg gcagctcccc
      481 tacccccctt acctctctag tctgtgctag ctcttccagc cccctgtcat ggcatcttcc
      541 aggggtccga gagctcagct agtcttcttc ctccaacccg ggcccctatg tccacttcag
      601 gacagcatgt ttgctgcctc cagggatcct gtgtccccga gctgggacca ccttatattc
      661 ccagggccgg ttaatgtggc tctggttctg ggtactttta tctgtcccct ccaccccaca
      721 gtggggccac tagggacacg cgtgttaaca agcttaggat tggtgacaga aaagccccat
      781 ccttaggcct cctccttcct agtctcctga tattgggtct aacccccacc tcctgttagg
      841 cagattcctt atctggtgac acacccccat ttcctggagc catctctctc cttgccagaa
      901 cctctaaggt ttgcttacga tggagccaga gaggatcctg ggagggagag cttggcaggg
      961 ggtgggaggg aaggggggga tgcgtgacct gcccggttct cagtggccac cctgcgctac
     1021 cctctcccag aacctgagct gctctgacgc ggccatcgga tcccgggccc gtcgactgca
     1081 gaggcctgca tgcaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     1141 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     1201 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     1261 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     1321 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     1381 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     1441 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     1501 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     1561 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     1621 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     1681 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     1741 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     1801 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     1861 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     1921 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     1981 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     2041 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     2101 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     2161 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     2221 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     2281 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     2341 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     2401 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     2461 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     2521 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     2581 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     2641 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     2701 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     2761 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     2821 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     2881 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     2941 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     3001 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     3061 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     3121 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     3181 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     3241 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     3301 ataaaaatag gcgtatcacg aggccctttc gtc
//
