LOCUS       sequence-275817-        4199 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Drug Resistance pDONR221HYG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4199)
  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 4199)
  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: pGL2207
FEATURES             Location/Qualifiers
     source          1..4199
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(151..178)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(270..356)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     405..427
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     419..436
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     420..436
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    452..551
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             847..1872
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             847..1872
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(2014..2025)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(2207..2306)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2323..2342)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2324..2342)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2347..2363)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2347..2363)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2465..2484)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2476..3285
                     /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(2543..2562)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3432..4020
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3921..3940
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4174..4191
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
       61 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atacgcgtac
      121 cgctagccag gaagagtttg tagaaacgca aaaaggccat ccgtcaggat ggccttctgc
      181 ttagtttgat gcctggcagt ttatggcggg cgtcctgccc gccaccctcc gggccgttgc
      241 ttcacaacgt tcaaatccgc tcccggcgga tttgtcctac tcaggagagc gttcaccgac
      301 aaacaacaga taaaacgaaa ggcccagtct tccgactgag cctttcgttt tatttgatgc
      361 ctggcagttc cctactctcg cgttaacgct agcatggatg ttttcccagt cacgacgttg
      421 taaaacgacg gccagtctta agctcgggcc ccaaataatg attttatttt gactgatagt
      481 gacctgttcg ttgcaacaaa ttgatgagca atgctttttt ataatgccaa ctttgtacaa
      541 aaaagcaggc tgtacacacg tacacacacg tgcgcatgct tgacgcatac gcgacgaatt
      601 cgaaagctca cctcattcct ccctcctcac accatcatcg gcatccatag agacacgtgc
      661 gcgtagaacg atacagctca cgcgcacaga gaggcgtgtt ctggtcgtgc gcatcatcgg
      721 cacagcactg gcggacgaaa ctcgcacaca ggcacgccgc ctcctttcac ccgtcataga
      781 tagttgaatt agacgccctc ctcctccctc atcatcgccg tcgtcatccg ggtccgagca
      841 ctagtgatga aaaagcctga actcaccgcg acgtctgtcg agaagtttct gatcgaaaag
      901 ttcgacagcg tctccgacct gatgcagctc tcggagggcg aagaatctcg tgctttcagc
      961 ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata gctgcgccga tggtttctac
     1021 aaagatcgtt atgtttatcg gcactttgca tcggccgcgc tcccgattcc ggaagtgctt
     1081 gacattgggg aattcagcga gagcctgacc tattgcatct cccgccgtgc acagggtgtc
     1141 acgttgcaag acctgcctga aaccgaactg cccgctgttc tgcagccggt cgcggaggcc
     1201 atggatgcga tcgctgcggc cgatcttagc cagacgagcg ggttcggccc attcggaccg
     1261 caaggaatcg gtcaatacac tacatggcgt gatttcatat gcgcgattgc tgatccccat
     1321 gtgtatcact ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc gcaggctctc
     1381 gatgagctga tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat
     1441 ttcggctcca acaatgtcct gacggacaat ggccgcataa cagcggtcat tgactggagc
     1501 gaggcgatgt tcggggattc ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg
     1561 ttggcttgta tggagcagca gacgcgctac ttcgagcgga ggcatccgga gcttgcagga
     1621 tcgccgcggc tccgggcgta tatgctccgc attggtcttg accaactcta tcagagcttg
     1681 gttgacggca atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga
     1741 tccggagccg ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc
     1801 gatggctgtg tagaagtact cgccgatagt ggaaaccgac gccccagcac tcgtccgagg
     1861 gcaaaggaat agagtagatg ccgaccggga tcctctagta gtgatcccgc ccgcacccgc
     1921 gcgcgctcag gccgcgtgtc gtcgtgctct ttccattttt tttttgtgtg tggtggtgat
     1981 ttgagctgct ctccgttgtg tgctggggac cctccttccc tcgatctcct cgtgcggggt
     2041 ctccgacgcc gcagacgcgg tgcgtgcgag cgtgcacgtg gctgtcgccg tggctgtagt
     2101 tgcgagcgga gagagagaga gagggaggga gggagggcag agggcagagg acatgcgggt
     2161 gggaacgtgc accggcctcg tctcacgcag ctggagccca cgaatgaccc agctttcttg
     2221 tacaaagttg gcattataag aaagcattgc ttatcaattt gttgcaacga acaggtcact
     2281 atcagtcaaa ataaaatcat tatttgccat ccagctgata tcccctatag tgagtcgtat
     2341 tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac
     2401 attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt
     2461 aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg attaaattcc
     2521 aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt
     2581 gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct gaaacatggc
     2641 aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa
     2701 tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc
     2761 accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc tgattcaggt
     2821 gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt
     2881 aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat
     2941 aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa
     3001 gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt
     3061 gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt
     3121 ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt
     3181 gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat
     3241 atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga attggttaat
     3301 tggttgtaac actggcagag cattacgctg acttgacggg acggcgcaag ctcatgacca
     3361 aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc
     3421 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     3481 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     3541 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     3601 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     3661 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     3721 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     3781 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     3841 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     3901 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     3961 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     4021 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     4081 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     4141 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcgga
//
