LOCUS       sequence-262346-        6825 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Leishmania donovani ectodomain in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6825)
  AUTHORS   Roberts AJ, Ong HB, Clare S, Brandt C, Harcourt K, Franssen SU, 
            Cotton JA, Muller-Sienerth N, Wright GJ
  TITLE     Systematic identification of genes encoding cell surface and 
            secreted proteins that are essential for in vitro growth and 
            infection in Leishmania donovani.
  JOURNAL   PLoS Pathog. 2022 Feb 24;18(2):e1010364. doi: 
            10.1371/journal.ppat.1010364. eCollection 2022 Feb.
  PUBMED    35202447
REFERENCE   2  (bases 1 to 6825)
  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: "PLoS 
            Pathog."; date: "2022-02-24"; volume: "18(2):e1010364. doi"; pages: 
            " 10.1371/journal.ppat.1010364. eCollection 2022 Feb"
COMMENT     Sequence Label: D54_ Ld_061160
FEATURES             Location/Qualifiers
     source          1..6825
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        7..386
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        387..590
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     541..561
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     587..611
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     1092..1111
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1134..1143
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1989..2006
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2058..2113
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2112..2131)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4349..4367)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4435..4539
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4540..4608,4609..5400)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4758..4777)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5571..6159
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6060..6079
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6313..6330
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6447..6468
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6483..6488,6489..6506,6507..6513)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6521..6537
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     6526..6548
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6545..6561
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6545..6561
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 catgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
       61 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      121 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      181 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      241 tacatcaagt gtatcatatg ccaagtccgc cccctattga cgtcaatgac ggtaaatggc
      301 ccgcctggca ttatgcccag tacatgacct tacgggactt tcctacttgg cagtacatct
      361 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacacc aatgggcgtg
      421 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      481 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taataacccc gccccgttga
      541 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga
      601 accgtcagat cctcactctc ttccgcatcg ctgtctgcga gggccagctg ttgggctcgc
      661 ggttgaggac aaactcttcg cggtctttcc agtactcttg gatcggaaac ccgtcggcct
      721 ccgaacggta ctccgccacc gagggacctg agcgagtccg catcgaccgg atcggaaaac
      781 ctctcgagaa aggcgtctaa ccagtcacag tcgcaaggta ggctgagcac cgtggcgggc
      841 ggcagcgggt ggcggtcggg gttgtttctg gcggaggtgc tgctgatgat gtaattaaag
      901 taggcggtct tgagacggcg gatggtcgag gtgaggtgtg gcaggcttga gatccagctg
      961 ttggggtgag tactccctct caaaagcggg cattacttct gcgctaagat tgtcagtttc
     1021 caaaaacgag gaggatttga tattcacctg gcccgatctg gccatacact tgagtgacaa
     1081 tgacatccac tttgcctttc tctccacagg tgtccactcc caggtccaag tttgccacca
     1141 tggagtttca gacccaggta ctcatgtccc tgctgctctg catgtctggt gcggccgccg
     1201 tgacagccgt gcagagcggc agctgcatct tcctgaccag cccccccagc tacaccttca
     1261 tcggcggagg cttccacatg cagctggccc tggaataccc cctgcccgcc aaggacgtgc
     1321 ggctgagctt cgacctgccc aagagcttct tcgtggacga ggccgaggcc gagcagctgt
     1381 accggatgga agtgctgagc gccgacccca tgacacaggg cacagccgcc gtggtggtgg
     1441 ccgacgtgac aagagcctac agccccctgc ggatgagcag cagatacgcc ttcgacatcg
     1501 aggcccccgt gttcaaagtg aactacacca ccaaccacgt ggaactgacc ttccagcagc
     1561 tgggcggcga cggcgctgga agcgccctgg atgcctatct ggccgaagat ggcgcccagg
     1621 cccaggtgcc cttccgggcc agactggtca tccccatcca ctccagatac gaggtgctgg
     1681 acaccaagac ccccttcagc ctgctgcggt tcatcaccgg cgagggcgcc tacgtgcggc
     1741 ggtgcctgac aagagtggat gtggccggca gagccgacag cagatgcctg accggcgcct
     1801 acagatccag cgccaccgtg gccaagagcg aggaaaagcc ccactacaag cactgcctgg
     1861 acctgcccgt gggcctgctg gctgatctgc ctggcgcgcc cggcagcctg caccacatcc
     1921 tggacgccca gaagatgctg tggaaccaca gggacaggaa cctgcctcct ctggcccctc
     1981 tgggccccca ccaccatcac caccattgag tctagacacg tgattaatta aggatccccc
     2041 gacctcgacc tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt
     2101 tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttggtcg agatccctcg
     2161 gagatctcta gctagaggat cgatccccgc cccggacgaa ctaaacctga ctacgacatc
     2221 tctgcccctt cttcgcgggg cagtgcatgt aatcccttca gttggttggt acaacttgcc
     2281 aactgggccc tgttccacat gtgacacggg gggggaccaa acacaaaggg gttctctgac
     2341 tgtagttgac atccttataa atggatgtgc acatttgcca acactgagtg gctttcatcc
     2401 tggagcagac tttgcagtct gtggactgca acacaacatt gcctttatgt gtaactcttg
     2461 gctgaagctc ttacaccaat gctgggggac atgtacctcc caggggccca ggaagactac
     2521 gggaggctac accaacgtca atcagagggg cctgtgtagc taccgataag cggaccctca
     2581 agagggcatt agcaatagtg tttataaggc ccccttgtta accctaaacg ggtagcatat
     2641 gcttcccggg tagtagtata tactatccag actaacccta attcaatagc atatgttacc
     2701 caacgggaag catatgctat cgaattaggg ttagtaaaag ggtcctaagg aacagcgata
     2761 tctcccaccc catgagctgt cacggtttta tttacatggg gtcaggattc cacgagggta
     2821 gtgaaccatt ttagtcacaa gggcagtggc tgaagatcaa ggagcgggca gtgaactctc
     2881 ctgaatcttc gcctgcttct tcattctcct tcgtttagct aatagaataa ctgctgagtt
     2941 gtgaacagta aggtgtatgt gaggtgctcg aaaacaaggt ttcaggtgac gcccccagaa
     3001 taaaatttgg acggggggtt cagtggtggc attgtgctat gacaccaata taaccctcac
     3061 aaaccccttg ggcaataaat actagtgtag gaatgaaaca ttctgaatat ctttaacaat
     3121 agaaatccat ggggtgggga caagccgtaa agactggatg tccatctcac acgaatttat
     3181 ggctatgggc aacacataat cctagtgcaa tatgatactg gggttattaa gatgtgtccc
     3241 aggcagggac caagacaggt gaaccatgtt gttacactct atttgtaaca aggggaaaga
     3301 gagtggacgc cgacagcagc ggactccact ggttgtctct aacacccccg aaaattaaac
     3361 ggggctccac gccaatgggg cccataaaca aagacaagtg gccactcttt tttttgaaat
     3421 tgtggagtgg gggcacgcgt cagcccccac acgccgccct gcggttttgg actgtaaaat
     3481 aagggtgtaa taacttggct gattgtaacc ccgctaacca ctgcggtcaa accacttgcc
     3541 cacaaaacca ctaatggcac cccggggaat acctgcataa gtaggtgggc gggccaagat
     3601 aggggcgcga ttgctgcgat ctggaggaca aattacacac acttgcgcct gagcgccaag
     3661 cacagggttg ttggtcctca tattcacgag gtcgctgaga gcacggtggg ctaatgttgc
     3721 catgggtagc atatactacc caaatatctg gatagcatat gctatcctaa tctatatctg
     3781 ggtagcatag gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg
     3841 ggtagtatat gctatcctaa tttatatctg ggtagcatag gctatcctaa tctatatctg
     3901 ggtagcatat gctatcctaa tctatatctg ggtagtatat gctatcctaa tctgtatccg
     3961 ggtagcatat gctatcctaa tagagattag ggtagtatat gctatcctaa tttatatctg
     4021 ggtagcatat actacccaaa tatctggata gcatatgcta tcctaatcta tatctgggta
     4081 gcatatgcta tcctaatcta tatctgggta gcataggcta tcctaatcta tatctgggta
     4141 gcatatgcta tcctaatcta tatctgggta gtatatgcta tcctaattta tatctgggta
     4201 gcataggcta tcctaatcta tatctgggta gcatatgcta tcctaatcta tatctgggta
     4261 gtatatgcta tcctaatctg tatccgggta gcatatgcta tcctcatgat aagctgtcaa
     4321 acatgagaat tttcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     4381 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4441 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4501 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     4561 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     4621 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     4681 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     4741 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag
     4801 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     4861 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     4921 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     4981 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     5041 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg
     5101 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     5161 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     5221 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     5281 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     5341 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     5401 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     5461 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5521 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5581 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5641 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5701 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     5761 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5821 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     5881 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     5941 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     6001 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6061 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6121 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6181 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     6241 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     6301 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     6361 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     6421 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     6481 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     6541 cacacaggaa acagctatga ccatgattac gccaagctct agctagaggt cgaccaattc
     6601 tcatgtttga cagcttatca tcgcagatcc gggcaacgtt gttgccattg ctgcaggcgc
     6661 agaactggta ggtatggaag atctatacat tgaatcaata ttggcaatta gccatattag
     6721 tcattggtta tatagcataa atcaatattg gctattggcc attgcatacg ttgtatctat
     6781 atcataatat gtacatttat attggctcat gtccaatatg accgc
//
