LOCUS       sequence-274684-        6306 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 6306)
  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 6306)
  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: D84_Ld_101170
FEATURES             Location/Qualifiers
     source          1..6306
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     201..225
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     706..725
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      748..757
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1084..1101
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1153..1208
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1207..1226)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3444..3462)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3530..3634
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3635..3703,3704..4495)
                     /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(3853..3872)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4666..5254
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5155..5174
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5408..5425
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5542..5563
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5578..5583,5584..5601,5602..5608)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5616..5632
                     /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     5621..5643
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5640..5656
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5640..5656
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        5927..6306
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta caccaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcctcac tctcttccgc
      241 atcgctgtct gcgagggcca gctgttgggc tcgcggttga ggacaaactc ttcgcggtct
      301 ttccagtact cttggatcgg aaacccgtcg gcctccgaac ggtactccgc caccgaggga
      361 cctgagcgag tccgcatcga ccggatcgga aaacctctcg agaaaggcgt ctaaccagtc
      421 acagtcgcaa ggtaggctga gcaccgtggc gggcggcagc gggtggcggt cggggttgtt
      481 tctggcggag gtgctgctga tgatgtaatt aaagtaggcg gtcttgagac ggcggatggt
      541 cgaggtgagg tgtggcaggc ttgagatcca gctgttgggg tgagtactcc ctctcaaaag
      601 cgggcattac ttctgcgcta agattgtcag tttccaaaaa cgaggaggat ttgatattca
      661 cctggcccga tctggccata cacttgagtg acaatgacat ccactttgcc tttctctcca
      721 caggtgtcca ctcccaggtc caagtttgcc accatggagt ttcagaccca ggtactcatg
      781 tccctgctgc tctgcatgtc tggtgcggcc gccagacgag ctgatgtgat ggttcgaaac
      841 gtgggcaccg aaaccaccaa cggcacactg ctggaactga gcaacgtgat ctgcaacgcc
      901 agatgcatca gcctgcctcg gctgtttcaa acccctcctg gacctgttct gccctctcct
      961 ccacacacct acaagcacag aagcgctggc gcgcccggca gcctgcacca catcctggac
     1021 gcccagaaga tgctgtggaa ccacagggac aggaacctgc ctcctctggc ccctctgggc
     1081 ccccaccacc atcaccacca ttgagtctag acacgtgatt aattaaggat cccccgacct
     1141 cgacctctgg ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt
     1201 gtctctcact cggaaggaca tatgggaggg caaatcattt ggtcgagatc cctcggagat
     1261 ctctagctag aggatcgatc cccgccccgg acgaactaaa cctgactacg acatctctgc
     1321 cccttcttcg cggggcagtg catgtaatcc cttcagttgg ttggtacaac ttgccaactg
     1381 ggccctgttc cacatgtgac acgggggggg accaaacaca aaggggttct ctgactgtag
     1441 ttgacatcct tataaatgga tgtgcacatt tgccaacact gagtggcttt catcctggag
     1501 cagactttgc agtctgtgga ctgcaacaca acattgcctt tatgtgtaac tcttggctga
     1561 agctcttaca ccaatgctgg gggacatgta cctcccaggg gcccaggaag actacgggag
     1621 gctacaccaa cgtcaatcag aggggcctgt gtagctaccg ataagcggac cctcaagagg
     1681 gcattagcaa tagtgtttat aaggccccct tgttaaccct aaacgggtag catatgcttc
     1741 ccgggtagta gtatatacta tccagactaa ccctaattca atagcatatg ttacccaacg
     1801 ggaagcatat gctatcgaat tagggttagt aaaagggtcc taaggaacag cgatatctcc
     1861 caccccatga gctgtcacgg ttttatttac atggggtcag gattccacga gggtagtgaa
     1921 ccattttagt cacaagggca gtggctgaag atcaaggagc gggcagtgaa ctctcctgaa
     1981 tcttcgcctg cttcttcatt ctccttcgtt tagctaatag aataactgct gagttgtgaa
     2041 cagtaaggtg tatgtgaggt gctcgaaaac aaggtttcag gtgacgcccc cagaataaaa
     2101 tttggacggg gggttcagtg gtggcattgt gctatgacac caatataacc ctcacaaacc
     2161 ccttgggcaa taaatactag tgtaggaatg aaacattctg aatatcttta acaatagaaa
     2221 tccatggggt ggggacaagc cgtaaagact ggatgtccat ctcacacgaa tttatggcta
     2281 tgggcaacac ataatcctag tgcaatatga tactggggtt attaagatgt gtcccaggca
     2341 gggaccaaga caggtgaacc atgttgttac actctatttg taacaagggg aaagagagtg
     2401 gacgccgaca gcagcggact ccactggttg tctctaacac ccccgaaaat taaacggggc
     2461 tccacgccaa tggggcccat aaacaaagac aagtggccac tctttttttt gaaattgtgg
     2521 agtgggggca cgcgtcagcc cccacacgcc gccctgcggt tttggactgt aaaataaggg
     2581 tgtaataact tggctgattg taaccccgct aaccactgcg gtcaaaccac ttgcccacaa
     2641 aaccactaat ggcaccccgg ggaatacctg cataagtagg tgggcgggcc aagatagggg
     2701 cgcgattgct gcgatctgga ggacaaatta cacacacttg cgcctgagcg ccaagcacag
     2761 ggttgttggt cctcatattc acgaggtcgc tgagagcacg gtgggctaat gttgccatgg
     2821 gtagcatata ctacccaaat atctggatag catatgctat cctaatctat atctgggtag
     2881 cataggctat cctaatctat atctgggtag catatgctat cctaatctat atctgggtag
     2941 tatatgctat cctaatttat atctgggtag cataggctat cctaatctat atctgggtag
     3001 catatgctat cctaatctat atctgggtag tatatgctat cctaatctgt atccgggtag
     3061 catatgctat cctaatagag attagggtag tatatgctat cctaatttat atctgggtag
     3121 catatactac ccaaatatct ggatagcata tgctatccta atctatatct gggtagcata
     3181 tgctatccta atctatatct gggtagcata ggctatccta atctatatct gggtagcata
     3241 tgctatccta atctatatct gggtagtata tgctatccta atttatatct gggtagcata
     3301 ggctatccta atctatatct gggtagcata tgctatccta atctatatct gggtagtata
     3361 tgctatccta atctgtatcc gggtagcata tgctatcctc atgataagct gtcaaacatg
     3421 agaattttct tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     3481 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     3541 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3601 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     3661 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     3721 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     3781 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     3841 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact
     3901 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     3961 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     4021 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     4081 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     4141 agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg
     4201 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     4261 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     4321 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     4381 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     4441 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     4501 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     4561 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     4621 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4681 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     4741 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     4801 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4861 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4921 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4981 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5041 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     5101 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     5161 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     5221 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     5281 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     5341 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     5401 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     5461 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     5521 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     5581 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     5641 aggaaacagc tatgaccatg attacgccaa gctctagcta gaggtcgacc aattctcatg
     5701 tttgacagct tatcatcgca gatccgggca acgttgttgc cattgctgca ggcgcagaac
     5761 tggtaggtat ggaagatcta tacattgaat caatattggc aattagccat attagtcatt
     5821 ggttatatag cataaatcaa tattggctat tggccattgc atacgttgta tctatatcat
     5881 aatatgtaca tttatattgg ctcatgtcca atatgaccgc catgttgaca ttgattattg
     5941 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
     6001 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
     6061 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
     6121 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
     6181 ccaagtccgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
     6241 tacatgacct tacgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
     6301 accatg
//
