LOCUS       sequence-193526-        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"
     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(832..849)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(832..848)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(841..863)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1057..1076)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1176..1198
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1236..1254)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1322..1426
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1427..1495,1496..2287)
                     /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(1645..1664)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2458..3046
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2947..2966
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3200..3217
                     /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 atggccgcgt
      181 cagagcagct caggttctgg gagagggtag cgcagggtgg ccactgagaa ccgggcaggt
      241 cacgcatccc ccccttccct cccaccccct gccaagctct ccctcccagg atcctctctg
      301 gctccatcgt aagcaaacct tagaggttct ggcaaggaga gagatggctc caggaaatgg
      361 gggtgtgtca ccagataagg aatctgccta acaggaggtg ggggttagac ccaatatcag
      421 gagactagga aggaggaggc ctaaggatgg ggcttttctg tcaccaatcc taagcttgtt
      481 aacacgcgtg tccctagtgg ccccactgtg gggtggaggg gacagataaa agtacccaga
      541 accagagcca cattaaccgg ccctgggaat ataaggtggt cccagctcgg ggacacagga
      601 tccctggagg cagcaaacat gctgtcctga agtggacata ggggcccggg ttggaggaag
      661 aagactagct gagctctcgg acccctggaa gatgccatga cagggggctg gaagagctag
      721 cacagactag agaggtaagg ggggtagggg agctgcccaa atgaaaggag tgagaggtga
      781 cccgaatccg cggccatcta gatgcattcg cgaggtaccg agctcgaatt cactggccgt
      841 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
      901 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
      961 acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc ttacgcatct
     1021 gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata
     1081 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     1141 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     1201 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     1261 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     1321 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     1381 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     1441 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     1501 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     1561 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     1621 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     1681 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     1741 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     1801 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     1861 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     1921 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     1981 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     2041 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     2101 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     2161 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     2221 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     2281 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     2341 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     2401 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     2461 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     2521 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     2581 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     2641 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     2701 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     2761 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     2821 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     2881 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     2941 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     3001 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     3061 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     3121 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     3181 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     3241 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     3301 cgactggaaa gcgggcagtg agcgcaacgc aat
//
