LOCUS       sequence-364348-        4252 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cloning plasmid for the generation of Knock-In in human cells using 
            rAAV..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4252)
  AUTHORS   Britton S, Coates J, Jackson SP
  TITLE     A new method for high-resolution imaging of Ku foci to decipher 
            mechanisms of DNA double-strand break repair.
  JOURNAL   J Cell Biol. 2013 Jul 29.
  PUBMED    23897892
REFERENCE   2  (bases 1 to 4252)
  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: "J Cell 
            Biol. 2013 Jul 29."
COMMENT     Sequence Label: pAAV-MCS2
FEATURES             Location/Qualifiers
     source          1..4252
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   56..196
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   67..196
                     /label=AAV2 ITR
     repeat_region   1581..1710
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(1772..2360)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1852..1871)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2531..3322,3323..3391))
                     /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     3154..3173
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3392..3496)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3564..3582
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3620..3642)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3742..3761
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(3778..4233)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3915..3936)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4127..4146
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgcccctgc
       61 aggcagctgc gcgctcgctc gctcactgag gccgcccggg caaagcccgg gcgtcgggcg
      121 acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc
      181 atcactaggg gttcctgcgg ccgcgctagc gaattcgcag ctgttgtgct gtgtatggac
      241 gtgggcttta ccatgagtaa ctccattcct ggtatagaat ccccatttga acaagcaaag
      301 aaggtgataa ccatgtttgt acagcgacag gtgtttgctg agaacaagga tgagattgct
      361 ttagtcctgt ttggtacaga tggcactgac aatccccttt ctggtgggga tcagtatcag
      421 aacatcacag tgcacagaca tctgatgcta ccagattttg atttgctgga ggacattgaa
      481 agcaaaatcc aaccaggttc tcaacaggct gacttcctgg atgcactaat cgtgagcatg
      541 gatgtgattc aacatgaaac aataggaaag aagtttgaga agaggcatat tgaaatattc
      601 actgacctca gcagccgatt cagcaaaagt cagctggata ttataattca tagcttgaag
      661 aaatgtggca tctccctgca attcttcttg cctttctcac ttggcaagga agatggaagt
      721 ggggacagag gagatggccc ctttcgctta ggtggccatg ggccttcctt tccactaaaa
      781 ggaattaccg aacagcaaaa agaaggtctt gagatagtga aaatggtgat gatatcttta
      841 gaaggtgaag atgggttgga tgaaatttat tcattcagtg agagtctgag aaaactgtgc
      901 gtcttcaaga aaattgagag gcattccatt cactggccct gccgactgac cattggctcc
      961 aatttgtcta taaggattgc agcctataaa tcgattctac aggagagagt taaaaagact
     1021 tggacagttg tggatgcaaa aaccctaaaa aaagaagata tacaaaaaga aacagtttat
     1081 tgcttaaatg atgatgatga aactgaagtt ttaaaagagg atattattca agggttccgc
     1141 tatggaagtg atatagttcc tttctctaaa gtggatgagg aacaaatgaa atataaatcg
     1201 gaggggaagt gcttctctgt tttgggattt tgtaaatctt ctcaggttca gagaagattc
     1261 ttcatgggaa atcaagttct aaaggtcttt gcagcaagag atgatgaggc agctgcagtt
     1321 gcactttcct ccctgattca tgctttggat gacttagaca tggtggccat agttcgatac
     1381 gcttatgaca aaagagctaa tcctcaagtc ggcgtggctt ttcctcatat caagcataac
     1441 tatgagtgtt tagtgtatgt gcagctgcct ttcatggaag acttgcggca atacatgttt
     1501 tcatccttga aaaacagtaa gaaatatgct cccaccgagg cacagttgaa tgctgttgat
     1561 gggatccacg cgtgggccgc aggaacccct agtgatggag ttggccactc cctctctgcg
     1621 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gcggcctcag
     1681 tgagcgagcg agcgcgcagc tgcctgcagg acatgtgagc aaaaggccag caaaaggcca
     1741 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     1801 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     1861 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     1921 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     1981 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     2041 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     2101 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     2161 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     2221 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     2281 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     2341 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     2401 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     2461 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     2521 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     2581 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     2641 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     2701 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     2761 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     2821 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     2881 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     2941 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     3001 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     3061 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     3121 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     3181 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     3241 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     3301 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     3361 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     3421 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     3481 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     3541 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt
     3601 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     3661 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     3721 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccataaaa
     3781 ttgtaaacgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     3841 ttaaccaata gaccgaaatc ggcaaaatcc cttataaatc aaaagaatag cccgagatag
     3901 agttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     3961 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcacccaaat
     4021 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     4081 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     4141 aaggagcggg cgctaaggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     4201 ccgccgcgct taatgcgccg ctacagggcg cgtactatgg ttgctttgac gt
//
