LOCUS       sequence-262310-        6480 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 6480)
  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 6480)
  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: D15_Ld_061240
FEATURES             Location/Qualifiers
     source          1..6480
                     /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             1644..1661
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1713..1768
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1767..1786)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4004..4022)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4090..4194
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4195..4263,4264..5055)
                     /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(4413..4432)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5226..5814
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5715..5734
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5968..5985
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6102..6123
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6138..6143,6144..6161,6162..6168)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6176..6192
                     /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     6181..6203
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6200..6216
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6200..6216
                     /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 ccgccgaagc ctctagagtg gtggaactga cccgggacac cttcgacgcc tacgtgggcg
     1261 gacccgacac ctacaccttt gtgctgtact gcgtgacctg gtcccggcag tgcaagaccg
     1321 ccatccagct gtgggaccgg ctgagcatca gccagagcca gaaagaactg cgggacgtgt
     1381 tcaccgccgc ctatgtggac ggcgacaagt accccgacct cgtgaagcgg atgaacgtga
     1441 agggcttccc caccatgctg ttctacaccc ccctgcacac cgagggcctc gagtacaacg
     1501 gccccagaga gagctacctg ctggacagct tcgtgttcca gttctctggc gcgcccggca
     1561 gcctgcacca catcctggac gcccagaaga tgctgtggaa ccacagggac aggaacctgc
     1621 ctcctctggc ccctctgggc ccccaccacc atcaccacca ttgagtctag acacgtgatt
     1681 aattaaggat cccccgacct cgacctctgg ctaataaagg aaatttattt tcattgcaat
     1741 agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg caaatcattt
     1801 ggtcgagatc cctcggagat ctctagctag aggatcgatc cccgccccgg acgaactaaa
     1861 cctgactacg acatctctgc cccttcttcg cggggcagtg catgtaatcc cttcagttgg
     1921 ttggtacaac ttgccaactg ggccctgttc cacatgtgac acgggggggg accaaacaca
     1981 aaggggttct ctgactgtag ttgacatcct tataaatgga tgtgcacatt tgccaacact
     2041 gagtggcttt catcctggag cagactttgc agtctgtgga ctgcaacaca acattgcctt
     2101 tatgtgtaac tcttggctga agctcttaca ccaatgctgg gggacatgta cctcccaggg
     2161 gcccaggaag actacgggag gctacaccaa cgtcaatcag aggggcctgt gtagctaccg
     2221 ataagcggac cctcaagagg gcattagcaa tagtgtttat aaggccccct tgttaaccct
     2281 aaacgggtag catatgcttc ccgggtagta gtatatacta tccagactaa ccctaattca
     2341 atagcatatg ttacccaacg ggaagcatat gctatcgaat tagggttagt aaaagggtcc
     2401 taaggaacag cgatatctcc caccccatga gctgtcacgg ttttatttac atggggtcag
     2461 gattccacga gggtagtgaa ccattttagt cacaagggca gtggctgaag atcaaggagc
     2521 gggcagtgaa ctctcctgaa tcttcgcctg cttcttcatt ctccttcgtt tagctaatag
     2581 aataactgct gagttgtgaa cagtaaggtg tatgtgaggt gctcgaaaac aaggtttcag
     2641 gtgacgcccc cagaataaaa tttggacggg gggttcagtg gtggcattgt gctatgacac
     2701 caatataacc ctcacaaacc ccttgggcaa taaatactag tgtaggaatg aaacattctg
     2761 aatatcttta acaatagaaa tccatggggt ggggacaagc cgtaaagact ggatgtccat
     2821 ctcacacgaa tttatggcta tgggcaacac ataatcctag tgcaatatga tactggggtt
     2881 attaagatgt gtcccaggca gggaccaaga caggtgaacc atgttgttac actctatttg
     2941 taacaagggg aaagagagtg gacgccgaca gcagcggact ccactggttg tctctaacac
     3001 ccccgaaaat taaacggggc tccacgccaa tggggcccat aaacaaagac aagtggccac
     3061 tctttttttt gaaattgtgg agtgggggca cgcgtcagcc cccacacgcc gccctgcggt
     3121 tttggactgt aaaataaggg tgtaataact tggctgattg taaccccgct aaccactgcg
     3181 gtcaaaccac ttgcccacaa aaccactaat ggcaccccgg ggaatacctg cataagtagg
     3241 tgggcgggcc aagatagggg cgcgattgct gcgatctgga ggacaaatta cacacacttg
     3301 cgcctgagcg ccaagcacag ggttgttggt cctcatattc acgaggtcgc tgagagcacg
     3361 gtgggctaat gttgccatgg gtagcatata ctacccaaat atctggatag catatgctat
     3421 cctaatctat atctgggtag cataggctat cctaatctat atctgggtag catatgctat
     3481 cctaatctat atctgggtag tatatgctat cctaatttat atctgggtag cataggctat
     3541 cctaatctat atctgggtag catatgctat cctaatctat atctgggtag tatatgctat
     3601 cctaatctgt atccgggtag catatgctat cctaatagag attagggtag tatatgctat
     3661 cctaatttat atctgggtag catatactac ccaaatatct ggatagcata tgctatccta
     3721 atctatatct gggtagcata tgctatccta atctatatct gggtagcata ggctatccta
     3781 atctatatct gggtagcata tgctatccta atctatatct gggtagtata tgctatccta
     3841 atttatatct gggtagcata ggctatccta atctatatct gggtagcata tgctatccta
     3901 atctatatct gggtagtata tgctatccta atctgtatcc gggtagcata tgctatcctc
     3961 atgataagct gtcaaacatg agaattttct tgaagacgaa agggcctcgt gatacgccta
     4021 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     4081 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     4141 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     4201 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     4261 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     4321 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     4381 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt
     4441 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     4501 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     4561 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     4621 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     4681 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca
     4741 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     4801 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     4861 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     4921 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     4981 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     5041 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     5101 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     5161 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5221 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     5281 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     5341 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     5401 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     5461 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     5521 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     5581 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     5641 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     5701 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     5761 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     5821 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     5881 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     5941 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     6001 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     6061 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     6121 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     6181 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gctctagcta
     6241 gaggtcgacc aattctcatg tttgacagct tatcatcgca gatccgggca acgttgttgc
     6301 cattgctgca ggcgcagaac tggtaggtat ggaagatcta tacattgaat caatattggc
     6361 aattagccat attagtcatt ggttatatag cataaatcaa tattggctat tggccattgc
     6421 atacgttgta tctatatcat aatatgtaca tttatattgg ctcatgtcca atatgaccgc
//
