LOCUS       sequence-118375-        3108 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bleomycin drug selection marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3108)
  AUTHORS   Zhang WW, Matlashewski G
  TITLE     CRISPR-Cas9-Mediated Genome Editing in Leishmania donovani.
  JOURNAL   MBio. 2015 Jul 21;6(4). pii: e00861-15. doi: 10.1128/mBio.00861-15.
  PUBMED    26199327
REFERENCE   2  (bases 1 to 3108)
  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: "MBio."; 
            date: "2015-07-21"; volume: "6(4). pii: e00861-15. doi"; pages: " 
            10.1128/mBio.00861-15"
COMMENT     Sequence Label: pSPble
FEATURES             Location/Qualifiers
     source          1..3108
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             301..675
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     primer_bind     complement(745..764)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(746..764)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(851..868)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1022..1610)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1102..1121)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1781..2572,2573..2641))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2404..2423
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2642..2746)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2814..2832
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2870..2892)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2992..3011
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(3092..3108,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(3092..3108,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaactcgagc agctgaagct tgcatgcctg caggtcgact ctagaggatc cccggccgcc
       61 ttgtgcggct cagcggaggc gctgagtgcg agcaccgccg cgacgcacac cagcaacacc
      121 acaagcggac gcacgctgcg gatcttcatc ggatcgggta ccgagctctc tcttctcctt
      181 cccctctctc tcctctctcc tctctccaga gctctctctt ctccttcccc tctctctcct
      241 ctctcctctc tccagagctc gaattcatcg atgatatcag atccccaact agatatcacc
      301 atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc
      361 gagttctgga ccgaccggct cgggttctcc cgggacttcg tggaggacga cttcgccggt
      421 gtggtccggg acgacgtgac cctgttcatc agcgcggtcc aggaccaggt ggtgccggac
      481 aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag
      541 gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca tgaccgagat cggcgagcag
      601 ccgtgggggc gggagttcgc cctgcgcgac ccggccggca actgcgtgca cttcgtggcc
      661 gaggagcagg actgaccgac gccgaccaac accgccggtc cgacgcggcc cgacgggtcc
      721 gaggcctcgg agatctgccg gtctccctat agtgagtcgt attaatttcg ataagccagg
      781 ttaacctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
      841 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
      901 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      961 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     1021 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     1081 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     1141 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     1201 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     1261 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     1321 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     1381 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     1441 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     1501 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     1561 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     1621 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     1681 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     1741 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     1801 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1861 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1921 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1981 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     2041 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     2101 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     2161 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     2221 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     2281 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     2341 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     2401 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     2461 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     2521 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     2581 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     2641 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     2701 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     2761 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     2821 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     2881 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     2941 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     3001 agcagattgt actgagagtg caccatatgg acatattgtc gttagaacgc ggctacaatt
     3061 aatacataac cttatgtatc atacacatac gatttaggtg acactata
//
