LOCUS       sequence-262306-        7140 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 7140)
  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 7140)
  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: D11_Ld_211610
FEATURES             Location/Qualifiers
     source          1..7140
                     /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             2304..2321
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2373..2428
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2427..2446)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4664..4682)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4750..4854
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4855..4923,4924..5715)
                     /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(5073..5092)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5886..6474
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6375..6394
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6628..6645
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6762..6783
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6798..6803,6804..6821,6822..6828)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6836..6852
                     /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     6841..6863
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6860..6876
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6860..6876
                     /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 gcggccgcca
     1201 ccgctcctgg ccctagcagc ttcagatgcc tgcggccttt cagcgccacc ggcaccttca
     1261 ccaacgtggc caaggcccac cccacccaga actgcgacta cttctacttc agcgtggaag
     1321 atgccaaggc cgagctgaag tgcgccagcc cttctgccgc caccaccgcc gtgtccgacg
     1381 agatgagctg caacgtgacc gtgcggatga gcatgcggag agccagcgac gtggtgcccc
     1441 ggctgcagag acaggtcgga accgacaagg actacttcac ccacgaggcc agcaccgaca
     1501 gcaccaccgc cagccccacc agagtgggca ataccagagc ctccgccgtg cccttcctga
     1561 ccgaggacga ggatgtgtgg cgctacgccg tgtccctgaa ggccaagggc gacagcagca
     1621 tccactacaa gggcttcaac ggcggcaacg gctacagcct gtgctgtgcc gccctggaag
     1681 aggccgactg cgcctggatg accccccaga agggcgacaa ggacctggac tgcgacgagg
     1741 acgtggacgt ccgctttgcc cctcagcggg gcagagtgct gagaggctgc cccctgcctt
     1801 tcccagcccc tagatctgcc ggcgacggcg tgttcggagc ctctgctggc gccagagtgc
     1861 tgctgaccga cctggatgag ggcgagggct ctggcgtgtt ccacgccgtg atcaccaagc
     1921 ccctgcacag attcgtggaa ggcccctggg aagtgatggt gcagatgtgg cggcggagac
     1981 agcacatgcc tagagtgggc cgcgaggaca gcctgagcgt gcccgccgat cacagcatcg
     2041 aggccgaggt gctgggccgg atcatggtgc ccttcaccct gaacctgacc gagctgcaga
     2101 agcagggcag aatcacccag atgcccagca tggccctgac agtggaagat gtggccgacg
     2161 tggcccaggc cgctggcgag aaagctgccg atgaagccgg cgacctgggc gcgcccggca
     2221 gcctgcacca catcctggac gcccagaaga tgctgtggaa ccacagggac aggaacctgc
     2281 ctcctctggc ccctctgggc ccccaccacc atcaccacca ttgagtctag acacgtgatt
     2341 aattaaggat cccccgacct cgacctctgg ctaataaagg aaatttattt tcattgcaat
     2401 agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg caaatcattt
     2461 ggtcgagatc cctcggagat ctctagctag aggatcgatc cccgccccgg acgaactaaa
     2521 cctgactacg acatctctgc cccttcttcg cggggcagtg catgtaatcc cttcagttgg
     2581 ttggtacaac ttgccaactg ggccctgttc cacatgtgac acgggggggg accaaacaca
     2641 aaggggttct ctgactgtag ttgacatcct tataaatgga tgtgcacatt tgccaacact
     2701 gagtggcttt catcctggag cagactttgc agtctgtgga ctgcaacaca acattgcctt
     2761 tatgtgtaac tcttggctga agctcttaca ccaatgctgg gggacatgta cctcccaggg
     2821 gcccaggaag actacgggag gctacaccaa cgtcaatcag aggggcctgt gtagctaccg
     2881 ataagcggac cctcaagagg gcattagcaa tagtgtttat aaggccccct tgttaaccct
     2941 aaacgggtag catatgcttc ccgggtagta gtatatacta tccagactaa ccctaattca
     3001 atagcatatg ttacccaacg ggaagcatat gctatcgaat tagggttagt aaaagggtcc
     3061 taaggaacag cgatatctcc caccccatga gctgtcacgg ttttatttac atggggtcag
     3121 gattccacga gggtagtgaa ccattttagt cacaagggca gtggctgaag atcaaggagc
     3181 gggcagtgaa ctctcctgaa tcttcgcctg cttcttcatt ctccttcgtt tagctaatag
     3241 aataactgct gagttgtgaa cagtaaggtg tatgtgaggt gctcgaaaac aaggtttcag
     3301 gtgacgcccc cagaataaaa tttggacggg gggttcagtg gtggcattgt gctatgacac
     3361 caatataacc ctcacaaacc ccttgggcaa taaatactag tgtaggaatg aaacattctg
     3421 aatatcttta acaatagaaa tccatggggt ggggacaagc cgtaaagact ggatgtccat
     3481 ctcacacgaa tttatggcta tgggcaacac ataatcctag tgcaatatga tactggggtt
     3541 attaagatgt gtcccaggca gggaccaaga caggtgaacc atgttgttac actctatttg
     3601 taacaagggg aaagagagtg gacgccgaca gcagcggact ccactggttg tctctaacac
     3661 ccccgaaaat taaacggggc tccacgccaa tggggcccat aaacaaagac aagtggccac
     3721 tctttttttt gaaattgtgg agtgggggca cgcgtcagcc cccacacgcc gccctgcggt
     3781 tttggactgt aaaataaggg tgtaataact tggctgattg taaccccgct aaccactgcg
     3841 gtcaaaccac ttgcccacaa aaccactaat ggcaccccgg ggaatacctg cataagtagg
     3901 tgggcgggcc aagatagggg cgcgattgct gcgatctgga ggacaaatta cacacacttg
     3961 cgcctgagcg ccaagcacag ggttgttggt cctcatattc acgaggtcgc tgagagcacg
     4021 gtgggctaat gttgccatgg gtagcatata ctacccaaat atctggatag catatgctat
     4081 cctaatctat atctgggtag cataggctat cctaatctat atctgggtag catatgctat
     4141 cctaatctat atctgggtag tatatgctat cctaatttat atctgggtag cataggctat
     4201 cctaatctat atctgggtag catatgctat cctaatctat atctgggtag tatatgctat
     4261 cctaatctgt atccgggtag catatgctat cctaatagag attagggtag tatatgctat
     4321 cctaatttat atctgggtag catatactac ccaaatatct ggatagcata tgctatccta
     4381 atctatatct gggtagcata tgctatccta atctatatct gggtagcata ggctatccta
     4441 atctatatct gggtagcata tgctatccta atctatatct gggtagtata tgctatccta
     4501 atttatatct gggtagcata ggctatccta atctatatct gggtagcata tgctatccta
     4561 atctatatct gggtagtata tgctatccta atctgtatcc gggtagcata tgctatcctc
     4621 atgataagct gtcaaacatg agaattttct tgaagacgaa agggcctcgt gatacgccta
     4681 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     4741 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     4801 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     4861 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     4921 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     4981 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     5041 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt
     5101 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     5161 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     5221 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     5281 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     5341 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca
     5401 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     5461 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     5521 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     5581 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     5641 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     5701 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     5761 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     5821 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5881 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     5941 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     6001 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     6061 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     6121 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6181 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     6241 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     6301 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6361 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6421 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     6481 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     6541 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     6601 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     6661 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     6721 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     6781 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     6841 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gctctagcta
     6901 gaggtcgacc aattctcatg tttgacagct tatcatcgca gatccgggca acgttgttgc
     6961 cattgctgca ggcgcagaac tggtaggtat ggaagatcta tacattgaat caatattggc
     7021 aattagccat attagtcatt ggttatatag cataaatcaa tattggctat tggccattgc
     7081 atacgttgta tctatatcat aatatgtaca tttatattgg ctcatgtcca atatgaccgc
//
