LOCUS       sequence-178057-        2829 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for ObLiGaRe/ZFN mediated targeting to AAVS1 locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2829)
  AUTHORS   Pinto R, Hansen L, Hintze J, Almeida R, Larsen S, Coskun M, Davidsen
            J, Mitchelmore C, David L, Troelsen JT, Bennett EP
  TITLE     Precise integration of inducible transcriptional elements (PrIITE) 
            enables absolute control of gene expression.
  JOURNAL   Nucleic Acids Res. 2017 May 4. doi: 10.1093/nar/gkx371.
  PUBMED    28472465
REFERENCE   2  (bases 1 to 2829)
  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: "Nucleic 
            Acids Res. 2017 May 4. doi: 10.1093/nar/gkx371."
COMMENT     Sequence Label: pAAVS1 ObLiGaRe Donor vector/EPB58
FEATURES             Location/Qualifiers
     source          1..2829
                     /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"
     promoter        complement(447..465)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(448..465)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(653..669)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(653..669)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(666..688)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    677..693
                     /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(701..707,708..725,726..731))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    746..767
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(884..901)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1055..1643)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1135..1154)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1814..2629)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1814..2629)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2537..2556
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2630..2734)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2802..2820
                     /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 gacgcgtatt ggaagcttat
      421 cctgtcccta ggccacccac tgtgggtcta tagtgtcacc taaatcttaa ttaagatatc
      481 gtttaaacgt cgacgctagc aggccggcgg ataactagct gatcgcggtc ttccccgacc
      541 caggtcactt ctgggttata gctgagactc cggacagcat gcaaccatcc tgtccctagg
      601 ccacccactg tgggtaagct tccaatggcg cgccgagctt ggctcgagca tggtcatagc
      661 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
      721 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
      781 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
      841 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
      901 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
      961 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     1021 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     1081 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     1141 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     1201 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     1261 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     1321 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     1381 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     1441 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     1501 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     1561 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     1621 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     1681 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     1741 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     1801 cttggtctga cagttagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat
     1861 caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac
     1921 cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa
     1981 catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac
     2041 catgagtgac gactgaatcc ggtgagaatg gcaaaagttt atgcatttct ttccagactt
     2101 gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat
     2161 tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac
     2221 aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac
     2281 ctgaatcagg atattcttct aatacctgga atgctgtttt cccagggatc gcagtggtga
     2341 gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt
     2401 ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc
     2461 catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac
     2521 ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg
     2581 aatttaatcg cggcctagag caagacgttt cccgttgaat atggctcata ctcttccttt
     2641 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     2701 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     2761 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     2821 cctttcgtc
//
