LOCUS       sequence-230446-        4796 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SapTrap donor vector for point mutagenesis; 
            pENTR-221::syntron(loxp:unr-119+).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4796)
  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 4796)
  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: pMLS253
FEATURES             Location/Qualifiers
     source          1..4796
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(919..946)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1038..1124)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1173..1195
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1187..1204
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1188..1204
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1220..1319
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    1351..1384
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             3348..3359
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    3508..3541
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(3572..3671)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3688..3707)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3689..3707)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3712..3728)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3712..3728)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3830..3849)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3841..4650
                     /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(3908..3927)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg ctagccagga
      901 agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt agtttgatgc
      961 ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt cacaacgttc
     1021 aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata
     1081 aaacgaaagg cccagtcttc cgactgagcc tttcgtttta tttgatgcct ggcagttccc
     1141 tactctcgcg ttaacgctag catggatgtt ttcccagtca cgacgttgta aaacgacggc
     1201 cagtcttaag ctcgggcccc aaataatgat tttattttga ctgatagtga cctgttcgtt
     1261 gcaacaaatt gatgagcaat gcttttttat aatgccaact ttgtacaaaa aagcaggctg
     1321 ctcttcgagt ttaaacatga ttttactaac ataacttcgt ataatgtatg ctatacgaag
     1381 ttatttctag acattctcta atgaaaaaat ctttcagttg aaattgaaaa tgagttaaag
     1441 ttggagtttt tattgaaaac agatttccgt gtgattagtg tttttagcga gtgtgacagg
     1501 acagcgaaaa aatatagaaa caagggggga actgaaaagc ttaggaatgc attgaacatg
     1561 agaaggggaa agggaaggaa caaactagac aggaattatt ggaatttaat cacatttgga
     1621 gttttttttc tattcgacag aataattatc cagaacattt ttgtattaaa tatttatgca
     1681 tcatatgagt agtcggcttt gttgtgcatg acgagtttgt tatcgacgaa atagaagctg
     1741 tcagaacgag tctcgtttgg attgttgatc atgtcgtcca ctgaaaaaga gattagtctt
     1801 ttgaattgta ctttttagaa taatgactca ctgagttgtt gagagagttg agggaactca
     1861 tagatatgtt cacagttgtt tcgtgaattc ggaatacaga atccgaattc aaagtcaaaa
     1921 cacttcagaa ggcgatcttt gaagaagtga cgttcgatca ttcggaaatg atggattggg
     1981 atgtctccta ctttgaattc cacagttgct ccgaccgttt tgagtttcaa aaagtttgga
     2041 gcaaatctat aacgcacgta tcttgccgac tcctgtggcg actcatcatt ctcctgatcg
     2101 ttctccggtt tggcgatctc gaaaagcact tgctcagtgt ccagatcacg gatttggaac
     2161 ttggtgaact cgatgttata gatgttcgca gatggggagc ataagaatcc taaatttatg
     2221 ttttaaactg aaatccaaag ggagcaagat accttgagtg attcccggaa gtgctaaaac
     2281 gtcgttcgga gtgatttgag ctttcttcgc aagctccgat tccgttgtga ttccttgttc
     2341 ggtgcttggt ggtggccgtg gcatctggaa atatggaaaa gttcaacaaa aagaaaagag
     2401 aaaagaatga aatcggatat caagagttag ttgagcggtt tctctagttt tctgagtctc
     2461 acctgcgacg ggaaggtcgc cgagccgggt ggaatcgatc gttgttgctc ggctttcata
     2521 tcggtttggt tggaagcggc tgaaaacgga aagaagtgga agaaggaaaa gagtgtggtg
     2581 tgacaggaaa atggtaatta gagggtgcca aataaccagc tatattttgt ttttttttga
     2641 aaacattttt aaaaagaaaa atacgataat gatatcagat ggatttccgg aaaactggta
     2701 tgaaaaattt caaccttttt gagtacatgt aatcaaaata cactttgtaa attatcattt
     2761 ttattgaaac tccaccattt ttctatttat aacgctaata atttgaaaaa gaaacctatt
     2821 gcgaaccgcg gggtgaatcc caaaaacgaa tgcgttttgg tggagtgatt gattcgaatc
     2881 gaagaagaaa aagaagaaga cgtggaatag agagctcact cttaaccgag cagcacacac
     2941 cgacagaaaa aaaaatgaaa tgaatgaggg tcttcttctt cttcttcttc gaatgattga
     3001 cagaaatggg aaaaagagga agattgagaa gggaaaaagg aaggagaaaa gaagcagaag
     3061 aagacgtcag agaggagagg aacgagcgga aaagcagcgg gcgcaagtca tagaagtagc
     3121 agagctgggg agaagaagac actatccaag aaaggaatga cgagagagta tgcaaagggg
     3181 tatagggtgc agacagaata ggaacagaat aacagatgat gagccaagaa gagttgaaaa
     3241 gggcgatgaa tttgtcatgt aacttaattt gggtcaattt gagcatgatg aattgaaatc
     3301 atcccttgtt gggagttaat aaccggtttg ttatcagaaa ccctgtaata gaagggcgcc
     3361 ctaactttga gccaattcat cccggtttct gtcaaatata tcaaaaagtg gtcaactgac
     3421 aaattgtttt tgatattata ataaacattt tatccgttaa caattttcga atacttttta
     3481 caaggacttg gataaattgg ctcaaagata acttcgtata atgtatgcta tacgaagtta
     3541 ttaactaatc tgatttaaat tttcgaagag cacccagctt tcttgtacaa agttggcatt
     3601 ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag tcaaaataaa
     3661 atcattattt gccatccagc tgatatcccc tatagtgagt cgtattacat ggtcatagct
     3721 gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagataaa
     3781 aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt
     3841 atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat ggatgctgat
     3901 ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcgc
     3961 ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc
     4021 aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg
     4081 accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc
     4141 ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat
     4201 gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac
     4261 agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat
     4321 gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg
     4381 cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat
     4441 aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc
     4501 gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca
     4561 ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag
     4621 tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacactgg
     4681 cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc ccttaacgtg
     4741 agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttc
//
