LOCUS       sequence-274876-        6879 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 6879)
  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 6879)
  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: D67_Ld_111030
FEATURES             Location/Qualifiers
     source          1..6879
                     /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             2037..2054
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2106..2161
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2160..2179)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4397..4415)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4483..4587
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4588..4656,4657..5448)
                     /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(4806..4825)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5619..6207
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6108..6127
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6361..6378
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6495..6516
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6531..6536,6537..6554,6555..6561)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6569..6585
                     /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     6574..6596
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6593..6609
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6593..6609
                     /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 tctggcgggg 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 gccgcaagcg
     1201 caatggtcag taaagcaaca ggcggagcag ttaaagcgat gcaggctacc agtgctgcgt
     1261 ccaatgctat cgtcgttggt atgcaagccc cgttgggcgt gtatgcttgc gtggctgaac
     1321 cgccgagcac agcctctggg ccctatgctg gggttaaaac agtcgttgtt cgagccattt
     1381 tgcctcggcg ggccccggat actatgcagg tgcgagatat cctggacgta tatcccgcct
     1441 ctggcatcac ctgcgagaag gaaacggcag aagctgtgga gaactttaac agggcagcca
     1501 aagtcgctgt agagaaagct aaagctatga aagctactag ggtgaccctg gtgatgaagc
     1561 ccgctacgaa gtatgagaga ctgaatggct tgtttcggga aacctgcacg aaaactatcg
     1621 aggctgcagg gctctcagtg gagacttcca caacagctgc agcaaccaat actctcatta
     1681 tgttccctga aaagatgtct gtagtgatgg tgagtgatga tcccgtatgt gagaacgttc
     1741 agtatgcgtt cgccggggtc gtaggcggag tgcacaccac ttactatacc gatgctgggt
     1801 ctaagattca tggtggacac agttataaat ccgtagctat ggccctcgca gaggaactca
     1861 gatctctggg catgaaggct gaagcagcca aagtggaagc cgccgctcag aagtctccca
     1921 ggaacgcagc tgcggctatg ggcgcgcccg gcagcctgca ccacatcctg gacgcccaga
     1981 agatgctgtg gaaccacagg gacaggaacc tgcctcctct ggcccctctg ggcccccacc
     2041 accatcacca ccattgagtc tagacacgtg attaattaag gatcccccga cctcgacctc
     2101 tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc
     2161 actcggaagg acatatggga gggcaaatca tttggtcgag atccctcgga gatctctagc
     2221 tagaggatcg atccccgccc cggacgaact aaacctgact acgacatctc tgccccttct
     2281 tcgcggggca gtgcatgtaa tcccttcagt tggttggtac aacttgccaa ctgggccctg
     2341 ttccacatgt gacacggggg gggaccaaac acaaaggggt tctctgactg tagttgacat
     2401 ccttataaat ggatgtgcac atttgccaac actgagtggc tttcatcctg gagcagactt
     2461 tgcagtctgt ggactgcaac acaacattgc ctttatgtgt aactcttggc tgaagctctt
     2521 acaccaatgc tgggggacat gtacctccca ggggcccagg aagactacgg gaggctacac
     2581 caacgtcaat cagaggggcc tgtgtagcta ccgataagcg gaccctcaag agggcattag
     2641 caatagtgtt tataaggccc ccttgttaac cctaaacggg tagcatatgc ttcccgggta
     2701 gtagtatata ctatccagac taaccctaat tcaatagcat atgttaccca acgggaagca
     2761 tatgctatcg aattagggtt agtaaaaggg tcctaaggaa cagcgatatc tcccacccca
     2821 tgagctgtca cggttttatt tacatggggt caggattcca cgagggtagt gaaccatttt
     2881 agtcacaagg gcagtggctg aagatcaagg agcgggcagt gaactctcct gaatcttcgc
     2941 ctgcttcttc attctccttc gtttagctaa tagaataact gctgagttgt gaacagtaag
     3001 gtgtatgtga ggtgctcgaa aacaaggttt caggtgacgc ccccagaata aaatttggac
     3061 ggggggttca gtggtggcat tgtgctatga caccaatata accctcacaa accccttggg
     3121 caataaatac tagtgtagga atgaaacatt ctgaatatct ttaacaatag aaatccatgg
     3181 ggtggggaca agccgtaaag actggatgtc catctcacac gaatttatgg ctatgggcaa
     3241 cacataatcc tagtgcaata tgatactggg gttattaaga tgtgtcccag gcagggacca
     3301 agacaggtga accatgttgt tacactctat ttgtaacaag gggaaagaga gtggacgccg
     3361 acagcagcgg actccactgg ttgtctctaa cacccccgaa aattaaacgg ggctccacgc
     3421 caatggggcc cataaacaaa gacaagtggc cactcttttt tttgaaattg tggagtgggg
     3481 gcacgcgtca gcccccacac gccgccctgc ggttttggac tgtaaaataa gggtgtaata
     3541 acttggctga ttgtaacccc gctaaccact gcggtcaaac cacttgccca caaaaccact
     3601 aatggcaccc cggggaatac ctgcataagt aggtgggcgg gccaagatag gggcgcgatt
     3661 gctgcgatct ggaggacaaa ttacacacac ttgcgcctga gcgccaagca cagggttgtt
     3721 ggtcctcata ttcacgaggt cgctgagagc acggtgggct aatgttgcca tgggtagcat
     3781 atactaccca aatatctgga tagcatatgc tatcctaatc tatatctggg tagcataggc
     3841 tatcctaatc tatatctggg tagcatatgc tatcctaatc tatatctggg tagtatatgc
     3901 tatcctaatt tatatctggg tagcataggc tatcctaatc tatatctggg tagcatatgc
     3961 tatcctaatc tatatctggg tagtatatgc tatcctaatc tgtatccggg tagcatatgc
     4021 tatcctaata gagattaggg tagtatatgc tatcctaatt tatatctggg tagcatatac
     4081 tacccaaata tctggatagc atatgctatc ctaatctata tctgggtagc atatgctatc
     4141 ctaatctata tctgggtagc ataggctatc ctaatctata tctgggtagc atatgctatc
     4201 ctaatctata tctgggtagt atatgctatc ctaatttata tctgggtagc ataggctatc
     4261 ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt atatgctatc
     4321 ctaatctgta tccgggtagc atatgctatc ctcatgataa gctgtcaaac atgagaattt
     4381 tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     4441 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     4501 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     4561 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     4621 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     4681 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     4741 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     4801 gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc
     4861 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     4921 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     4981 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     5041 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     5101 ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta
     5161 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     5221 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     5281 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     5341 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     5401 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     5461 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     5521 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5581 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     5641 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     5701 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     5761 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     5821 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     5881 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     5941 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     6001 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     6061 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     6121 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     6181 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     6241 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     6301 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     6361 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     6421 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     6481 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
     6541 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     6601 agctatgacc atgattacgc caagctctag ctagaggtcg accaattctc atgtttgaca
     6661 gcttatcatc gcagatccgg gcaacgttgt tgccattgct gcaggcgcag aactggtagg
     6721 tatggaagat ctatacattg aatcaatatt ggcaattagc catattagtc attggttata
     6781 tagcataaat caatattggc tattggccat tgcatacgtt gtatctatat cataatatgt
     6841 acatttatat tggctcatgt ccaatatgac cgccatgtt
//
