LOCUS       sequence-275797-        7915 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of rapamycin-inducible dimerisation of split Cre from 
            ribosomal locus in Leishmania parasite; pRIB-DiCre.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7915)
  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 7915)
  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: pGL2399
FEATURES             Location/Qualifiers
     source          1..7915
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     21..43
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     40..56
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     40..56
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    76..96
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    361..385
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             597..617
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             624..944
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             2151..2171
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2178..2453
                     /codon_start=1
                     /product="T2098L mutant of FKBP-rapamycin binding domain of
                     human FRAP"
                     /label=FRB*
                     /note="binds synthetic ligands such as AP21967"
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLLQAWDLYYHVFRRIS"
     CDS             3864..4262
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             complement(4386..4397)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(4577..4601)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5567..5587)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     complement(5595..5612)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5595..5611)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5604..5626)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5939..5961
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5999..6017)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6085..6189
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6190..6258,6259..7050)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGSDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6408..6427)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7221..7809
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7710..7729
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctttgagtg agctgatacc agcggataac aatttcacac aggaaacagc tatgaccatg
       61 attacgccaa gctatcaact ttgtatagaa aagttgcctt aattaatgac gaacaactgc
      121 cctatcagct ggtgatggcc gtgtagtgga ctgccatggc gttgacggga gcgggggatt
      181 agggttcgat tccggagagg gagcctgaga aatagctacc acttctacgg agggcagcag
      241 gcgcgcaaat tgcccaatgt caaaacaaaa cgatgaggca gcgaaaagaa atagagttgt
      301 cagtccattt ggattgtcat ttcaatgggg gatatttaaa cccatccaat atcgagtaac
      361 gcaagtttgt acaaaaaagc aggcttgccc tcgaggtttt cgtcgcgcat cggtgtaggt
      421 cgaataccct tgcctttcag tgcggggttt tttcctataa agcatttatt gtctcgacgc
      481 ttctcctgct tggttctctc tcgcacttgc atgacagatt tcaagatatc ttgcctgtac
      541 actcgtcatt aaaaacagct tattcccggt tatttttctc aacgaagaaa atggccccta
      601 agaagaagag aaaggtgtca agaggagtgc aggtggaaac catctcccca ggagacgggc
      661 gcaccttccc caagcgcggc cagacctgcg tggtgcacta caccgggatg cttgaagatg
      721 gaaagaaatt tgattcctcc agggacagaa acaagccctt taagtttatg ctaggcaagc
      781 aggaggtgat ccgaggctgg gaagaagggg ttgcccagat gagtgtgggt cagagagcca
      841 aactgactat atctccagat tatgcctatg gtgccactgg gcacccaggc atcatcccac
      901 cacatgccac tctcgtcttc gatgtggagc ttctaaaact ggaagctagc cccagcaacc
      961 ccggcgccag caacggctcc acctctgatg aagtcaggaa gaacctgatg gacatgttca
     1021 gggacaggca ggccttctct gaacacacct ggaagatgct cctgtctgtg tgcagatcct
     1081 gggctgcctg gtgcaagctg aactgaaggt cctcacccga ctgggcagct ggagtgcaga
     1141 gaactttgcg cgaccggaag acgctagtga gattgggcac agtggagcta gcgtgcgcgc
     1201 ggtagctttg gcagcactgg gattttgtcc tctgtactgc ctggctctcg tgtttatttt
     1261 tggttagagt gtgcgttgct cggatccctg cctggttctc gtgcttattt ttggttagat
     1321 ggtgcgttgt tctactttca atcatcgttt tgttttgttt ttcatatctg ttttccaaag
     1381 aactctgatt gtcttcatcc acccggtttt ccccgcctgg tctcgctttg agattgtttt
     1441 tattcctctc cggtttttcc ccattgggat aaaagtgcgt cccgtgtgtc tacccctcca
     1501 gtccccacaa gcggctcaga tgtacagagc acctgctctt ctctgacgtg cactttttgc
     1561 tcttgcagtc ttgcgcggcc tctagtctcc cctgccacgc acggatgctg acgccaagca
     1621 gcattctgcg caggaaagcg tggtatcccg acgaggcaga tcgcgggctc gcatgtaccg
     1681 gcgacggaca tcctacacac attatcctca tgtagaacct caaagcaagg gaagaagaaa
     1741 agggttgaca cgttgttatg tttttttttg tgcctgggag taagcgggcg ccgatcttca
     1801 agtatcctct caatttcact ggcagggcgc gaagtgcgcg ggctcctcgc gtgccgccgc
     1861 aatgtcagtc gcggctctac cagtgctaca gggatgtcgg gtggtgtcgt tgatgatttg
     1921 gcatcaagtg cacgcgcggc cgctttcttc catctgcact cgctctgcgc ttcgttttct
     1981 tcttcccttg cgcgcgcgtg acgccgcgct cctcacctgc gcgccccgcc ccgacttggg
     2041 tcctctcgtg ctgcaacgga cgccgacagc ttccttccac gcatttcgtg cctccgcacc
     2101 gttctccacc gccagcgcag acttgtacat acgcacacac caccatggcc cctaagaaga
     2161 agagaaaggt gtcaagaatc ctctggcatg agatgtggca tgaaggcctg gaagaggcat
     2221 ctcgtttgta ctttggggaa aggaacgtga aaggcatgtt tgaggtgctg gagcccttgc
     2281 atgctatgat ggaacggggc ccccagactc tgaaggaaac atcctttaat caggcctatg
     2341 gtcgagattt aatggaggcc caagagtggt gcaggaagta catgaaatca gggaatgtca
     2401 aggacctcct ccaagcctgg gacctctatt atcatgtgtt ccgacgaatc tcagctagcc
     2461 ccagcaaccc cggcgccagc aacggctcca acaggaaatg gttccctgct gaacctgagg
     2521 atgtgaggga ctacctcctg tacctgcaag ccagaggcct ggctgtgaag accatccaac
     2581 agcacctggg ccagctcaac atgctgcaca ggaggtctgg cctgcctcgc ccttctgact
     2641 ccaatgctgt gtccctggtg atgaggagaa tcagaaagga gaatgtggat gctggggaga
     2701 gagccaagca ggccctggcc tttgaacgca ctgactttga ccaagtcaga tccctgatgg
     2761 agaactctga cagatgccag gacatcagga acctggcctt cctgggcatt gcctacaaca
     2821 ccctgctgcg cattgccgaa attgccagaa tcagagtgaa ggacatctcc cgcaccgatg
     2881 gtgggagaat gctgatccac attggcagga ccaagaccct ggtgtccaca gctggtgtgg
     2941 agaaggccct gtccctgggg gttaccaagc tggtggagag atggatctct gtgtctggtg
     3001 tggctgatga ccccaacaac tacctgttct gccgggtcag aaagaatggt gtggctgccc
     3061 cttctgccac ctcccaactg tccacccgag ccctggaagg gatctttgag gccacccacc
     3121 gcctgatcta tggtgccaag gatgactctg ggcagagata cctggcctgg tctggccact
     3181 ctgccagagt gggtgctgcc agggacatgg ccagggctgg tgtgtccatc cctgaaatca
     3241 tgcaggctgg tggctggacc aatgtgaaca ttgtgatgaa ctacatcaga aacctggact
     3301 ctgagactgg ggccatggtg aggctgctag aggatgggga ctgaagtgtg cgtgagtgtg
     3361 cgcgtgcgtg ttggtgcgca ccgcgctcgc tgtccccgcg gcgcacgcac acctccctcg
     3421 ctctctcccc ctctttccgt gccttctttg tctcccttcc tgtacgctat tttggtttct
     3481 tgtgcttgct ttatcttgtg ggcgggtgtg ggcccgacct gcacgtctgt gacggagtga
     3541 cgtctagaga gctcgggtcg accacacgta cacacgtaca cacacacgtg cgcatgcttg
     3601 acgcatacgc gacgaattcg aaagctcacc tcattcctcc ctcctcacac catcatcggc
     3661 atccatagag acacgtgcgc gtagaacgat acagctcacg cgcacagaga ggcgtgttct
     3721 ggtcgtgcgc atcatcggca cagcactggc ggacgaaact cgcacacagg cacgccgcct
     3781 cctttcaccc gtcatagata gttgaattag acgccctcct cctccctcat catcgccgtc
     3841 gtcatccggg tccgagcact agtatggcca agcctttgtc tcaagaagaa tccaccctca
     3901 ttgaaagagc aacggctaca atcaacagca tccccatctc tgaagactac agcgtcgcca
     3961 gcgcagctct ctctagcgac ggccgcatct tcactggtgt caatgtatat cattttactg
     4021 ggggaccttg tgcagaactc gtggtgctgg gcactgctgc tgctgcggca gctggcaacc
     4081 tgacttgtat cgtcgcgatc ggaaatgaga acaggggcat cttgagcccc tgcggacggt
     4141 gccgacaggt gcttctcgat ctgcatcctg ggatcaaagc catagtgaag gacagtgatg
     4201 gacagccgac ggcagttggg attcgtgaat tgctgccctc tggttatgtg tgggagggct
     4261 aaagatcttc tagtgatccc gcccgcaccc gcgcgcgctc aggccgcgtg tcgtcgtgct
     4321 ctttccattt tttttttgtg tgtggtggtg atttgagctg ctctccgttg tgtgctgggg
     4381 accctccttc cctcgatctc ctcgtgcggg gtctccgacg cgcagacgcg gtgcgtgcga
     4441 gcgtgcacgt gctgtcgccg tggctgtagt tgcgagcgga gagagagaga gagggaggga
     4501 gggagggcag agggcagagg acatgcgggt gggaacgtgc accggcctcg tctcacgcag
     4561 gcccgggcgg taccctaccc agctttcttg tacaaagtgg ggcgactaga ccgtaacgcc
     4621 ttttcaactc acggcctcta ggaatgaagg agggtagttc gggggagaac gtactggggc
     4681 gtcagaggtg aaattcttag accgcaccaa gacgaactac agcgaaggca ttcttcaagg
     4741 ataccttcct caatcaagaa ccaaagtgtg gagatcgaag atgattagag accattgtag
     4801 tccacactgc aaacgatgac acccatgaat tggggatctt atgggccggc ctgcggcagg
     4861 gtttaccctg tgtcagcacc gcgcccgctt ttaccaactt acgtatcttt tctattcggc
     4921 ctttaccggc cacccacggg aatatcctca gcacgttttc tgttttttca cgcgaaagct
     4981 ttgaggttac agtctcaggg gggagtacgt tcgcaagagt gaaacttaaa gaaattgacg
     5041 gaatggcacc acaagacgtg gagcgtgcgg tttaatttga ctcaacacgg ggaactttac
     5101 cagatccgga caggatgagg attgacagat tgagtgttct ttctcgattc cctgaatggt
     5161 ggtgcatggc cgcttttggt cggtggagtg atttgtttgg ttgattccgt caacggacga
     5221 gatccaagct gcccagtaga attcagaatt gcccatagga tagcaaactc atcggcgggt
     5281 tttacccaac ggtgggccgc attcggtcga attcttctct gcgggattcc tttgtaattg
     5341 cacaaggtga aattttgggc aacagcaggt ctgtgatgct cctcaatgtt ctgggcgaca
     5401 cgcgcactac aatgtcagtg agaacaagaa aaacgacttt tgtcgaacct acttgatcaa
     5461 aagagtgggg aaaccccgga atcacataga cccacttggg accgaggatt gcaattattg
     5521 gtcgcgcaac gaggaatgtc tcgtaggcgc agctcatgtt taaacccaac tttattatac
     5581 atagttgata attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     5641 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     5701 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcctgatg
     5761 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt
     5821 gcaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     5881 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     5941 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     6001 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     6061 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     6121 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     6181 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     6241 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     6301 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     6361 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     6421 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     6481 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     6541 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     6601 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     6661 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     6721 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     6781 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     6841 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     6901 cgtgggtctc gcggtatcat tgcagcactg gggtcagatg gtaagccctc ccgtatcgta
     6961 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     7021 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     7081 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     7141 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     7201 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     7261 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     7321 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     7381 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     7441 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     7501 agacgatagt taccggataa ggcgcagcga tcgggctgaa cggggggttc gtgcacacag
     7561 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     7621 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     7681 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     7741 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     7801 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggtcttttg ctggcctttt
     7861 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccg
//
