LOCUS       sequence-178611-        3556 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Codon-optimized HaloTag donor for the SapTrap cloning system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3556)
  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 3556)
  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: pDD378
FEATURES             Location/Qualifiers
     source          1..3556
                     /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"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1681..1780)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1797..1816)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1798..1816)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1821..1837)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1821..1837)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1939..1958)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1950..2759
                     /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(2017..2036)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2906..3494
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3395..3414
                     /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 agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctgc tcttcgatgg ctgaaattgg cacaggattc ccgtttgacc cccactacgt
      721 cgaggtcctc ggagagcgta tgcactacgt cgacgtcgga ccacgtgacg gaaccccagt
      781 cctcttcctc cacggaaacc caacctcctc ctacgtctgg cgtaacatca tcccacacgt
      841 cgccccaacc caccgttgca tcgccccaga cctcatcgga atgggaaagt ccgacaagcc
      901 agacctcgga tacttcttcg acgaccacgt ccgtttcatg gacgccttca tcgaggccct
      961 cggactcgag gaggtcgtcc tcgtcatcca cgactgggga tccgccctcg gattccactg
     1021 ggccaagcgt aacccagagc gtgtcaaggt aagtttaaac atatatatac taactaaccc
     1081 tgattattta aattttcagg gaatcgcctt catggagttc atccgtccaa tcccaacctg
     1141 ggacgagtgg ccagagttcg cccgtgagac cttccaagcc ttccgtacca ccgacgtcgg
     1201 acgtaagctc atcatcgacc aaaacgtctt catcgaggga accctcccaa tgggagtcgt
     1261 ccgtccactc accgaggtcg agatggacca ctaccgtgag ccattcctca acccagtcga
     1321 ccgtgagcca ctctggcgtt tcccaaacga gctcccaatc gccggagagc cagccaacat
     1381 cgtcgccctc gtcgaggagt acatggactg gctccaccaa tccccagtcc caaagctcct
     1441 cttctgggga accccaggag tcctcatccc accagccgag gccgcccgtc tcgccaagtc
     1501 cctcccaaac tgcaaggtaa gtttaaacag ttcggtacta actaaccata catatttaaa
     1561 ttttcaggcc gtcgacatcg gaccaggact caacctcctc caagaggaca acccagacct
     1621 catcggatcc gagatcgccc gttggctctc caccctcgag atctccggaa agcgaagagc
     1681 acccagcttt cttgtacaaa gttggcatta taagaaagca ttgcttatca atttgttgca
     1741 acgaacaggt cactatcagt caaaataaaa tcattatttg ccatccagct gatatcccct
     1801 atagtgagtc gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa
     1861 atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg
     1921 cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc
     1981 cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg
     2041 tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt
     2101 ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa
     2161 actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg
     2221 atgcatggtt actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat
     2281 atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt
     2341 cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc
     2401 aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct
     2461 ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag
     2521 tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag
     2581 gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat
     2641 ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta
     2701 ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat
     2761 cagaattggt taattggttg taacactggc agagcattac gctgacttga cgggacggcg
     2821 caagctcatg accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc
     2881 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     2941 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     3001 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     3061 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     3121 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     3181 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     3241 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     3301 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     3361 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     3421 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     3481 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     3541 cttttgctca catgtt
//
