LOCUS       sequence-178588-        4793 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pmyo-2::GFP FP-slot donor for the SapTrap cloning system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4793)
  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 4793)
  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: pDD401
FEATURES             Location/Qualifiers
     source          1..4793
                     /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"
     primer_bind     complement(1672..1700)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2448..2468
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     protein_bind    complement(2918..3017)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3034..3053)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3035..3053)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3058..3074)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3058..3074)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3176..3195)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3187..3996
                     /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(3254..3273)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4143..4731
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4632..4651
                     /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 tcttcgatga ttttatatct gagtagtatc ctttgcttta aatgtccata
      721 aaactaattt tataatcaat aaaacaacgt ttgtaaatca actgagttta caagtagaga
      781 cattgaggga tactttcact atgctaaagt gataatcgac caaataataa cttcactttg
      841 gtatttattc ctgtcttata aatgttatgt atgaattaaa ttcatatgca tatggctcac
      901 tctgacaatt tttaataatc ttccagatca atattgacta ccgatgcggg tggtcttttg
      961 ctttgaattc tgctgaactt tacaccccga acagcaatgt gtgcttcagc ctaaaaaaaa
     1021 gtaagtgtgt taatcagtgc ccccgattct tcattttttg cccctctctc ccgtttcgtc
     1081 ggcaaaagaa gagaaaataa agataagtct caagataggt tggtaatcgc taaagtggtt
     1141 gtgtggataa gagtagcaaa atggcagcaa gagcactttg cgcgcacaca ctgtactcat
     1201 tgttctggat aaaattctct cgttgtttgc cgtcggatgt ctgcctctct gcattgagcc
     1261 ggcttcttca ctatctttag ttaacctaaa atgccgtttc ttttctcgta tccccactat
     1321 cccgttgagg ttctctgctc tcttcgctcc ctaccgccag cgagcaacta tccgtggggg
     1381 cgccttgctc ggaagatggg ggggaagaaa gaagattttt gctatttgca cttgagaaag
     1441 agacttttcc tgcgtcgatg gttagagaac agtgtgcaga cacttttcag ctacctagaa
     1501 ttacaattgg atatccccgc ctcccaatcc acccacccag ggaaaaagaa gggctcgccg
     1561 aaaatcaaag ttatctccag gctcgcgcat cccaccgagc ggttgacttc tctccaccac
     1621 ttttcatttt taccctcgac catgagtaaa ggagaagaac ttttcactgg agttgtccca
     1681 attcttgttg aattagatgg tgatgttaat gggcacaaat tttctgtcag tggagagggt
     1741 gaaggtgatg caacatacgg aaaacttacc cttaaattta tttgcactac tggaaaacta
     1801 cctgttccat gggtaagttt aaacatatat atactaacta accctgatta tttaaatttt
     1861 cagccaacac ttgtcactac tttctgttat ggtgttcaat gcttctcgag atacccagat
     1921 catatgaaac ggcatgactt tttcaagagt gccatgcccg aaggttatgt acaggaaaga
     1981 actatatttt tcaaagatga cgggaactac aagacacgta agtttaaaca gttcggtact
     2041 aactaaccat acatatttaa attttcaggt gctgaagtca agtttgaagg tgataccctt
     2101 gttaatagaa tcgagttaaa aggtattgat tttaaagaag atggaaacat tcttggacac
     2161 aaattggaat acaactataa ctcacacaat gtatacatca tggcagacaa acaaaagaat
     2221 ggaatcaaag ttgtaagttt aaacatgatt ttactaacta actaatctga tttaaatttt
     2281 cagaacttca aaattagaca caacattgaa gatggaagcg ttcaactagc agaccattat
     2341 caacaaaata ctccaattgg cgatggccct gtccttttac cagacaacca ttacctgtcc
     2401 acacaatctg ccctttcgaa agatcccaac gaaaagagag accacatggt ccttcttgag
     2461 tttgtaacag ctgctgggat tacacatggc atggatgaac tatacaaata gatcaactgc
     2521 tctaattcca tctcgtttct ctcttttctt tcaatctctt tacccgtttc tcttccccta
     2581 cgtcccccaa acctaccact gcacagtaac acatacacat tcaacgtgtc acaataaaag
     2641 cccatcaaca ccttctctgt tgtttttttt cttcaaatgc atttttttca ctttgtttaa
     2701 taatttttac gcatcggatt tgtaggatag tggaaatttg tccttcaatg cttcaattga
     2761 ttcgacactt ctcaatattt ttgatcaata aaccattgat tttctattta gaagaagatc
     2821 actgtaaata acaccaaaaa catcacagca aaaaaactcc cactcaatta tattactgtt
     2881 tttattgaaa ctagaataaa tttgaaaagc gaagagcacc cagctttctt gtacaaagtt
     2941 ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa
     3001 aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta ttacatggtc
     3061 atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa
     3121 gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg
     3181 ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc caacatggat
     3241 gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc
     3301 tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc
     3361 gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct
     3421 cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg
     3481 atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt
     3541 gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct
     3601 tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg
     3661 gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa
     3721 gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca
     3781 cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc
     3841 ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct
     3901 ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa
     3961 ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa
     4021 cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc aaaatccctt
     4081 aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4141 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4201 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4261 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     4321 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4381 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4441 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     4501 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     4561 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4621 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     4681 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     4741 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gtt
//
