LOCUS       sequence-227770-        4791 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SapTrap donor vector for point mutagenesis; 
            pENTR-221::syntron(lox2272-119+).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4791)
  AUTHORS   Schwartz M, Jorgensen E
  TITLE     SapTrap vectors for introducing point mutations with unc-119+ 
            selection.
  JOURNAL   MicroPubl Biol. 2018 Jul 29;2018:10.17912/DDVH-BG64. doi: 
            10.17912/DDVH-BG64.
  PUBMED    32550388
REFERENCE   2  (bases 1 to 4791)
  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: "2018-07-29"; volume: "2018:10.17912/DDVH-BG64. doi"; 
            pages: " 10.17912/DDVH-BG64"
COMMENT     Sequence Label: pMLS415
FEATURES             Location/Qualifiers
     source          1..4791
                     /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    700..733
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     CDS             2692..2703
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    2852..2885
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(2916..3015)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3032..3051)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3033..3051)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3056..3072)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3056..3072)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3174..3193)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3185..3994
                     /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(3252..3271)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4141..4729
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4630..4649
                     /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 tcttcgagtt taaacatgat tttactaaca taacttcgta taaagtatcc
      721 tatacgaagt tatgacattc tctaatgaaa aaatctttca gttgaaattg aaaatgagtt
      781 aaagttggag tttttattga aaacagattt ccgtgtgatt agtgttttta gcgagtgtga
      841 caggacagcg aaaaaatata gaaacaaggg gggaactgaa aagcttagga atgcattgaa
      901 catgagaagg ggaaagggaa ggaacaaact agacaggaat tattggaatt taatcacatt
      961 tggagttttt tttctattcg acagaataat tatccagaac atttttgtat taaatattta
     1021 tgcatcatat gagtagtcgg ctttgttgtg catgacgagt ttgttatcga cgaaatagaa
     1081 gctgtcagaa cgagtctcgt ttggattgtt gatcatgtcg tccactgaaa aagagattag
     1141 tcttttgaat tgtacttttt agaataatga ctcactgagt tgttgagaga gttgagggaa
     1201 ctcatagata tgttcacagt tgtttcgtga attcggaata cagaatccga attcaaagtc
     1261 aaaacacttc agaaggcgat ctttgaagaa gtgacgttcg atcattcgga aatgatggat
     1321 tgggatgtct cctactttga attccacagt tgctccgacc gttttgagtt tcaaaaagtt
     1381 tggagcaaat ctataacgca cgtatcttgc cgactcctgt ggcgactcat cattctcctg
     1441 atcgttctcc ggtttggcga tctcgaaaag cacttgctca gtgtccagat cacggatttg
     1501 gaacttggtg aactcgatgt tatagatgtt cgcagatggg gagcataaga atcctaaatt
     1561 tatgttttaa actgaaatcc aaagggagca agataccttg agtgattccc ggaagtgcta
     1621 aaacgtcgtt cggagtgatt tgagctttct tcgcaagctc cgattccgtt gtgattcctt
     1681 gttcggtgct tggtggtggc cgtggcatct ggaaatatgg aaaagttcaa caaaaagaaa
     1741 agagaaaaga atgaaatcgg atatcaagag ttagttgagc ggtttctcta gttttctgag
     1801 tctcacctgc gacgggaagg tcgccgagcc gggtggaatc gatcgttgtt gctcggcttt
     1861 catatcggtt tggttggaag cggctgaaaa cggaaagaag tggaagaagg aaaagagtgt
     1921 ggtgtgacag gaaaatggta attagagggt gccaaataac cagctatatt ttgttttttt
     1981 ttgaaaacat ttttaaaaag aaaaatacga taatgatatc agatggattt ccggaaaact
     2041 ggtatgaaaa atttcaacct ttttgagtac atgtaatcaa aatacacttt gtaaattatc
     2101 atttttattg aaactccacc atttttctat ttataacgct aataatttga aaaagaaacc
     2161 tattgcgaac cgcggggtga atcccaaaaa cgaatgcgtt ttggtggagt gattgattcg
     2221 aatcgaagaa gaaaaagaag aagacgtgga atagagagct cactcttaac cgagcagcac
     2281 acaccgacag aaaaaaaaat gaaatgaatg agggtcttct tcttcttctt cttcgaatga
     2341 ttgacagaaa tgggaaaaag aggaagattg agaagggaaa aaggaaggag aaaagaagca
     2401 gaagaagacg tcagagagga gaggaacgag cggaaaagca gcgggcgcaa gtcatagaag
     2461 tagcagagct ggggagaaga agacactatc caagaaagga atgacgagag agtatgcaaa
     2521 ggggtatagg gtgcagacag aataggaaca gaataacaga tgatgagcca agaagagttg
     2581 aaaagggcga tgaatttgtc atgtaactta atttgggtca atttgagcat gatgaattga
     2641 aatcatccct tgttgggagt taataaccgg tttgttatca gaaaccctgt aatagaaggg
     2701 cgccctaact ttgagccaat tcatcccggt ttctgtcaaa tatatcaaaa agtggtcaac
     2761 tgacaaattg tttttgatat tataataaac attttatccg ttaacaattt tcgaatactt
     2821 tttacaagga cttggataaa ttggctcaaa gataacttcg tataaagtat cctatacgaa
     2881 gttattaact aatctgattt aaattttcga agagcaccca gctttcttgt acaaagttgg
     2941 cattataaga aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa
     3001 taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt acatggtcat
     3061 agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga
     3121 taaaaatata tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg
     3181 tgttatgagc catattcaac gggaaacgtc gaggccgcga ttaaattcca acatggatgc
     3241 tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta
     3301 tcgcttgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt
     3361 tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct
     3421 tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat
     3481 ccccggaaaa acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt
     3541 tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt
     3601 taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt
     3661 tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga
     3721 aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact
     3781 tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg
     3841 aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc
     3901 ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt
     3961 gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca
     4021 ctggcagagc attacgctga cttgacggga cggcgcaagc tcatgaccaa aatcccttaa
     4081 cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     4141 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     4201 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     4261 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     4321 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     4381 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     4441 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     4501 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     4561 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     4621 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     4681 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     4741 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt t
//
