LOCUS       sequence-178615-        2693 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AID FP-slot donor for the SapTrap cloning system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2693)
  AUTHORS   Dickinson D, Slabodnick M, Chen A, Goldstein B
  TITLE     SapTrap assembly of repair templates for Cas9-triggered homologous 
            recombination with a self-excising cassette.
  JOURNAL   MicroPubl Biol. 2018 May 1;2018:10.17912/W2KT0N. doi: 
            10.17912/W2KT0N.
  PUBMED    32550377
REFERENCE   2  (bases 1 to 2693)
  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: "MicroPubl 
            Biol."; date: "2018-05-1"; volume: "2018:10.17912/W2KT0N. doi"; 
            pages: " 10.17912/W2KT0N"
COMMENT     Sequence Label: pDD397
FEATURES             Location/Qualifiers
     source          1..2693
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     563..582
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        563..581
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     606..622
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     607..623
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    639..663
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             675..806
                     /codon_start=1
                     /product="minimal degron from the auxin-responsive IAA17 
                     protein of Arabidopsis thaliana (Morawska and Ulrich, 
                     2013)"
                     /label=AID*
                     /note="acts as an auxin-inducible degron (AID) in the 
                     presence of TIR1"
                     /translation="PKDPAKPPAKAQVVGWPPVRSYRKNVMVSCQKSSGGPEAAAFVK"
     protein_bind    complement(815..839)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(866..882)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(866..882)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(919..939)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(919..937)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(958..974)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(958..974)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1076..1095)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1087..1896
                     /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="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1154..1173)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2043..2631
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2532..2551
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtgagcgcg cgtaatacga ctcactatag ggcgaattgg gtaccgggcc
      601 ccccctcgag gtcgacggta tcgataagct tgatggggac aagtttgtac aaaaaagcag
      661 gctgctcttc gatgcctaaa gatccagcca aacctccggc caaggcacaa gttgtgggat
      721 ggccaccggt gagatcatac cggaagaacg tgatggtttc ctgccaaaaa tcaagcggtg
      781 gcccggaggc ggcggcgttc gtgaagcgaa gagcacccag ctttcttgta caaagtggtc
      841 cccatcgaat tcctgcagcc cgggggatcc actagttcta gagcggccgc caccgcggtg
      901 gagctccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc
      961 atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa
     1021 gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg
     1081 ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc caacatggat
     1141 gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc
     1201 tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc
     1261 gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct
     1321 cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg
     1381 atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt
     1441 gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct
     1501 tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg
     1561 gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa
     1621 gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca
     1681 cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc
     1741 ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct
     1801 ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa
     1861 ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa
     1921 cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc aaaatccctt
     1981 aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     2041 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     2101 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     2161 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     2221 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     2281 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     2341 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     2401 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     2461 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     2521 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     2581 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     2641 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gtt
//
