LOCUS       sequence-275795-        3606 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Drug Resistance pDONR221BSD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3606)
  AUTHORS   Duncan SM, Myburgh E, Alves-Ferreira EV, Mottram JC
  TITLE     DiCre-Based Inducible Disruption of Leishmania Genes.
  JOURNAL   Methods Mol Biol. 2019;1971:211-224. doi: 
            10.1007/978-1-4939-9210-2_10.
  PUBMED    30980305
REFERENCE   2  (bases 1 to 3606)
  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: "Methods Mol
            Biol."; date: "2019"; volume: "1971:211-224. doi"; pages: " 
            10.1007/978-1-4939-9210-2_10"
COMMENT     Sequence Label: pGL2208
FEATURES             Location/Qualifiers
     source          1..3606
                     /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"
     CDS             1013..1411
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             complement(1538..1549)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(1731..1830)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1847..1866)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1848..1866)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1871..1887)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1871..1887)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1989..2008)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2000..2809
                     /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(2067..2086)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2956..3544
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3445..3464
                     /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 agcaggctgt acacacgtac acacacacac acacacacac acacacacac acacacacgt
      721 acacacgtac acacacgtgc gcatgcttga cgcatacgcg acgaattcga aagctcacct
      781 cattcctccc tcctcacacc atcatcggca tccatagaga cacgtgcgcg tagaacgata
      841 cagctcacgc gcacagagag gcgtgttctg gtcgtgcgca tcatcggcac agcactggcg
      901 gacgaaactc gcacacaggc acgccgcctc ctttcacccg tcatagatag ttgaattaga
      961 cgccctcctc ctccctcatc atcgccgtcg tcatccgggt ccgagcacta gtatggccaa
     1021 gcctttgtct caagaagaat ccaccctcat tgaaagagca acggctacaa tcaacagcat
     1081 ccccatctct gaagactaca gcgtcgccag cgcagctctc tctagcgacg gccgcatctt
     1141 cactggtgtc aatgtatatc attttactgg gggaccttgt gcagaactcg tggtgctggg
     1201 cactgctgct gctgcggcag ctggcaacct gacttgtatc gtcgcgatcg gaaatgagaa
     1261 caggggcatc ttgagcccct gcggacggtg ccgacaggtg cttctcgatc tgcatcctgg
     1321 gatcaaagcc atagtgaagg acagtgatgg acagccgacg gcagttggga ttcgtgaatt
     1381 gctgccctct ggttatgtgt gggagggcta aggatcctct agtagtgatc ccgcccgcac
     1441 ccgcgcgcgc tcaggccgcg tgtcgtcgtg ctctttccat tttttttttg tgtgtggtgg
     1501 tgatttgagc tgctctccgt tgtgtgctgg ggaccctcct tccctcgatc tcctcgtgcg
     1561 gggtctccga cgccgcagac gcggtgcgtg cgagcgtgca cgtggctgtc gccgtggctg
     1621 tagttgcgag cggagagaga gagagaggga gggagggagg gcagagggca gaggacatgc
     1681 gggtgggaac gtgcaccggc ctcgtctcac gcagctggag cccacgaatg acccagcttt
     1741 cttgtacaaa gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt
     1801 cactatcagt caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc
     1861 gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg
     1921 ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa
     1981 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa
     2041 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     2101 aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     2161 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     2221 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     2281 actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc
     2341 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     2401 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat
     2461 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     2521 acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca
     2581 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     2641 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     2701 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     2761 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt
     2821 taattggttg taacactggc agagcattac gctgacttga cgggacggcg caagctcatg
     2881 accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa
     2941 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3001 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3061 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     3121 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3181 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3241 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3301 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     3361 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     3421 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     3481 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     3541 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     3601 catgtt
//
