LOCUS       sequence-183599-        3526 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 3526)
  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 3526)
  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: AX53_pS.Donor.S1.TS
FEATURES             Location/Qualifiers
     source          1..3526
                     /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(1305..1321)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1305..1321)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1318..1340)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1329..1345
                     /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(1353..1359,1360..1377,1378..1383))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1398..1419
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1536..1553)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1707..2295)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1787..1806)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2466..3257,3258..3326))
                     /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     3089..3108
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3327..3431)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3499..3517
                     /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 tatctaggga taacagggta atgtcgaggc cggatttaaa tctgtcccct
      481 ccaccccaca gtggggccac tagggacagg attggtgaca gagcggccgc ggattcgggt
      541 cacctctcac tcctttcatt tgggcagctc ccctaccccc cttacctctc tagtctgtgc
      601 tagctcttcc agccccctgt catggcatct tccaggggtc cgagagctca gctagtcttc
      661 ttcctccaac ccgggcccct atgtccactt caggacagca tgtttgctgc ctccagggat
      721 cctgtgtccc cgagctggga ccaccttata ttcccagggc cggttaatgt ggctctggtt
      781 ctgggtactt ttatctgtcc cctccacccc acagtggggc cactagggac acgcgtgtta
      841 acaagcttag gattggtgac agaaaagccc catccttagg cctcctcctt cctagtctcc
      901 tgatattggg tctaaccccc acctcctgtt aggcagattc cttatctggt gacacacccc
      961 catttcctgg agccatctct ctccttgcca gaacctctaa ggtttgctta cgatggagcc
     1021 agagaggatc ctgggaggga gagcttggca gggggtggga gggaaggggg ggatgcgtga
     1081 cctgcccggt tctcagtggc caccctgcgc taccctctcc cagaacctga gctgctctga
     1141 cgcggccgcg ccggcgctgt cccctccacc ccacagtggg gccactaggg acaggattgg
     1201 tgacagagtt taaactaggg ataacagggt aatgtcgagg ccgggatatc ggatcccggg
     1261 cccgtcgact gcagaggcct gcatgcaagc ttggcgtaat catggtcata gctgtttcct
     1321 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     1381 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     1441 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     1501 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     1561 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     1621 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     1681 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     1741 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     1801 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     1861 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     1921 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     1981 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     2041 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     2101 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     2161 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     2221 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     2281 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     2341 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     2401 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     2461 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     2521 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     2581 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     2641 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     2701 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     2761 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     2821 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     2881 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     2941 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     3001 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     3061 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     3121 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     3181 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     3241 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     3301 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     3361 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     3421 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     3481 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtc
//
