LOCUS       sequence-193528-        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"
     protein_bind    1..22
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(37..42,43..60,61..67)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    75..91
                     /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     80..102
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     99..115
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     99..115
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1025..1042)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1025..1041)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1034..1056)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1250..1269)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1369..1391
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1429..1447)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1515..1619
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1620..1688,1689..2480)
                     /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(1838..1857)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2651..3239
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3140..3159
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3393..3410
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
       61 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
      121 ttacgccaag cttgcatgca ggcctctgca gtcgacgggc ccgggatccg atatcccggc
      181 ctcgacatta ccctgttatc cctagtttaa actctgtcac caatcctgtc cctagtggcc
      241 ccactgtggg gtggagggga cagcgccggc gcggccgcgt cagagcagct caggttctgg
      301 gagagggtag cgcagggtgg ccactgagaa ccgggcaggt cacgcatccc ccccttccct
      361 cccaccccct gccaagctct ccctcccagg atcctctctg gctccatcgt aagcaaacct
      421 tagaggttct ggcaaggaga gagatggctc caggaaatgg gggtgtgtca ccagataagg
      481 aatctgccta acaggaggtg ggggttagac ccaatatcag gagactagga aggaggaggc
      541 ctaaggatgg ggcttttctg tcaccaatcc taagcttgtt aacacgcgtg tccctagtgg
      601 ccccactgtg gggtggaggg gacagataaa agtacccaga accagagcca cattaaccgg
      661 ccctgggaat ataaggtggt cccagctcgg ggacacagga tccctggagg cagcaaacat
      721 gctgtcctga agtggacata ggggcccggg ttggaggaag aagactagct gagctctcgg
      781 acccctggaa gatgccatga cagggggctg gaagagctag cacagactag agaggtaagg
      841 ggggtagggg agctgcccaa atgaaaggag tgagaggtga cccgaatccg cggccgctct
      901 gtcaccaatc ctgtccctag tggccccact gtggggtgga ggggacagat ttaaatccgg
      961 cctcgacatt accctgttat ccctagatat ctagatgcat tcgcgaggta ccgagctcga
     1021 attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
     1081 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
     1141 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcctgatg cggtattttc
     1201 tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct
     1261 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     1321 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     1381 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     1441 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     1501 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     1561 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     1621 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     1681 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     1741 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     1801 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     1861 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     1921 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     1981 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     2041 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     2101 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     2161 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     2221 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     2281 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     2341 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     2401 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     2461 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     2521 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     2581 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     2641 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     2701 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     2761 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     2821 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     2881 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     2941 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     3001 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     3061 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     3121 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     3181 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     3241 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     3301 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     3361 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     3421 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     3481 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaat
//
