LOCUS       sequence-274854-        6891 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 6891)
  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 6891)
  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: D75_Ld_302670
FEATURES             Location/Qualifiers
     source          1..6891
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     1086..1105
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1128..1137
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2049..2066
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2118..2173
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2172..2191)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4409..4427)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4495..4599
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4600..4668,4669..5460)
                     /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(4818..4837)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5631..6219
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6120..6139
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6373..6390
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6507..6528
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6543..6548,6549..6566,6567..6573)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6581..6597
                     /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     6586..6608
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6605..6621
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6605..6621
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agatcctcac tctcttccgc atcgctgtct gcgagggcca gctgttgggc tcgcggttga
      661 ggacaaactc ttcgcggtct ttccagtact cttggatcgg aaacccgtcg gcctccgaac
      721 ggtactccgc caccgaggga cctgagcgag tccgcatcga ccggatcgga aaacctctcg
      781 agaaaggcgt ctaaccagtc acagtcgcaa ggtaggctga gcaccgtggc gggcggcagc
      841 gggtggcggt cggggttgtt tctggcggag gtgctgctga tgatgtaatt aaagtaggcg
      901 gtcttgagac ggcggatggt cgaggtgagg tgtggcaggc ttgagatcca gctgttgggg
      961 tgagtactcc ctctcaaaag cgggcattac ttctgcgcta agattgtcag tttccaaaaa
     1021 cgaggaggat ttgatattca cctggcccga tctggccata cacttgagtg acaatgacat
     1081 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caagtttgcc accatggagt
     1141 ttcagaccca ggtactcatg tccctgctgc tctgcatgtc tggtgcggcc gccgaagtag
     1201 ttgtcgcccc cttgtgtgca gagaaacatg gtttcgccgg agaatttaac gatcattaca
     1261 ccataccagt agaacaggcg ccggctgctg gtgaggccat cgtgctcatc gcccgggcct
     1321 ttccactgaa tttgggaaat atgctcgcta ggtggctgta tatcaatgcc acctaccgga
     1381 tcagcgaagg cgcacaactt accgaggtct ccgacaataa cggtatcctt gagctggcca
     1441 acgtggctgc cggaaccgat atcgaggttc gagtaatgag agtgcgtaca gacgggccga
     1501 caccatttag gttcttcagc ttcttcgcca actctcctgt gtgccatgtt gcagtggcac
     1561 cagataactc tttcattgct cccgttccga ggcgtctgag agcgacccca cagttgcctt
     1621 tgacgatgta ctttaaggca gtgctgcccc cgaatgttaa cgcccttatc atccgagtaa
     1681 gcgcagacgg acgcaccgat gagaaattta gaagcggcga taaggtgtat gcttctggcg
     1741 acctggtccg cgcagcatca ggtggcgcag tgctgtttgc atatactttc cctatggaaa
     1801 acgctgccac tccctactac ttcacccaag tcagtgtgag ctatggcgag tgggaaggtg
     1861 gcgacgcggc caagccgtct gcatctaaca cgagttcatc tggcggtgac tctaaagaca
     1921 cgatgcaacc cacacgaagt acgcccgtgc ttggcgcgcc cggcagcctg caccacatcc
     1981 tggacgccca gaagatgctg tggaaccaca gggacaggaa cctgcctcct ctggcccctc
     2041 tgggccccca ccaccatcac caccattgag tctagacacg tgattaatta aggatccccc
     2101 gacctcgacc tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt
     2161 tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttggtcg agatccctcg
     2221 gagatctcta gctagaggat cgatccccgc cccggacgaa ctaaacctga ctacgacatc
     2281 tctgcccctt cttcgcgggg cagtgcatgt aatcccttca gttggttggt acaacttgcc
     2341 aactgggccc tgttccacat gtgacacggg gggggaccaa acacaaaggg gttctctgac
     2401 tgtagttgac atccttataa atggatgtgc acatttgcca acactgagtg gctttcatcc
     2461 tggagcagac tttgcagtct gtggactgca acacaacatt gcctttatgt gtaactcttg
     2521 gctgaagctc ttacaccaat gctgggggac atgtacctcc caggggccca ggaagactac
     2581 gggaggctac accaacgtca atcagagggg cctgtgtagc taccgataag cggaccctca
     2641 agagggcatt agcaatagtg tttataaggc ccccttgtta accctaaacg ggtagcatat
     2701 gcttcccggg tagtagtata tactatccag actaacccta attcaatagc atatgttacc
     2761 caacgggaag catatgctat cgaattaggg ttagtaaaag ggtcctaagg aacagcgata
     2821 tctcccaccc catgagctgt cacggtttta tttacatggg gtcaggattc cacgagggta
     2881 gtgaaccatt ttagtcacaa gggcagtggc tgaagatcaa ggagcgggca gtgaactctc
     2941 ctgaatcttc gcctgcttct tcattctcct tcgtttagct aatagaataa ctgctgagtt
     3001 gtgaacagta aggtgtatgt gaggtgctcg aaaacaaggt ttcaggtgac gcccccagaa
     3061 taaaatttgg acggggggtt cagtggtggc attgtgctat gacaccaata taaccctcac
     3121 aaaccccttg ggcaataaat actagtgtag gaatgaaaca ttctgaatat ctttaacaat
     3181 agaaatccat ggggtgggga caagccgtaa agactggatg tccatctcac acgaatttat
     3241 ggctatgggc aacacataat cctagtgcaa tatgatactg gggttattaa gatgtgtccc
     3301 aggcagggac caagacaggt gaaccatgtt gttacactct atttgtaaca aggggaaaga
     3361 gagtggacgc cgacagcagc ggactccact ggttgtctct aacacccccg aaaattaaac
     3421 ggggctccac gccaatgggg cccataaaca aagacaagtg gccactcttt tttttgaaat
     3481 tgtggagtgg gggcacgcgt cagcccccac acgccgccct gcggttttgg actgtaaaat
     3541 aagggtgtaa taacttggct gattgtaacc ccgctaacca ctgcggtcaa accacttgcc
     3601 cacaaaacca ctaatggcac cccggggaat acctgcataa gtaggtgggc gggccaagat
     3661 aggggcgcga ttgctgcgat ctggaggaca aattacacac acttgcgcct gagcgccaag
     3721 cacagggttg ttggtcctca tattcacgag gtcgctgaga gcacggtggg ctaatgttgc
     3781 catgggtagc atatactacc caaatatctg gatagcatat gctatcctaa tctatatctg
     3841 ggtagcatag gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg
     3901 ggtagtatat gctatcctaa tttatatctg ggtagcatag gctatcctaa tctatatctg
     3961 ggtagcatat gctatcctaa tctatatctg ggtagtatat gctatcctaa tctgtatccg
     4021 ggtagcatat gctatcctaa tagagattag ggtagtatat gctatcctaa tttatatctg
     4081 ggtagcatat actacccaaa tatctggata gcatatgcta tcctaatcta tatctgggta
     4141 gcatatgcta tcctaatcta tatctgggta gcataggcta tcctaatcta tatctgggta
     4201 gcatatgcta tcctaatcta tatctgggta gtatatgcta tcctaattta tatctgggta
     4261 gcataggcta tcctaatcta tatctgggta gcatatgcta tcctaatcta tatctgggta
     4321 gtatatgcta tcctaatctg tatccgggta gcatatgcta tcctcatgat aagctgtcaa
     4381 acatgagaat tttcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     4441 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4501 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4561 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     4621 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     4681 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     4741 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     4801 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag
     4861 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     4921 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     4981 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     5041 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     5101 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg
     5161 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     5221 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     5281 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     5341 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     5401 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     5461 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     5521 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5581 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5641 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5701 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5761 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     5821 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5881 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     5941 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     6001 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     6061 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6121 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6181 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6241 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     6301 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     6361 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     6421 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     6481 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     6541 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     6601 cacacaggaa acagctatga ccatgattac gccaagctct agctagaggt cgaccaattc
     6661 tcatgtttga cagcttatca tcgcagatcc gggcaacgtt gttgccattg ctgcaggcgc
     6721 agaactggta ggtatggaag atctatacat tgaatcaata ttggcaatta gccatattag
     6781 tcattggtta tatagcataa atcaatattg gctattggcc attgcatacg ttgtatctat
     6841 atcataatat gtacatttat attggctcat gtccaatatg accgccatgt t
//
