LOCUS       sequence-273949-        7656 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 7656)
  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 7656)
  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: D17_Ld_090870
FEATURES             Location/Qualifiers
     source          1..7656
                     /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             2814..2831
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2883..2938
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2937..2956)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5174..5192)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5260..5364
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5365..5433,5434..6225)
                     /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(5583..5602)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6396..6984
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6885..6904
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7138..7155
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7272..7293
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7308..7313,7314..7331,7332..7338)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7346..7362
                     /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     7351..7373
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7370..7386
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7370..7386
                     /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 gccaccggca
     1201 gacctaagca gcctatcagc aagggcacca ccgccggcac aagcagccac gccagcagca
     1261 gcctgtgccc cagatgcccc ctgtgcagca gaagcggcac ccccatcaaa gagcggctga
     1321 agtactgcca caccaccagc ggctacatca tcagcctgtt cagctggggc acctctgccc
     1381 tggccggcaa gcagagaacc ggcgtgtcca acagagagca gcggcagctg taccccctgg
     1441 aactgccctt cgccaccgac gacgccgtgc ggaagcagtg catgcgggac atgcaggacc
     1501 tgctgctggc cagagtgcgg ggccagcctc aggtggtcat gcccctgctg gacgtgctgc
     1561 ggagaaagct ggcctacccc cgcgagcctg tggtcgtgca cctggctggc gacaacggcg
     1621 tgggcaagac ccacaccgcc agactggtgt cccaggccct gagcctgaga tgcgccccag
     1681 acagagatgt gtgcgacgcc ggcgacaatc tgctgaccat tgccggcacc ggcttcgacg
     1741 gcctgtctgt ggccgaggcc cggcagagaa tcatcggcca gatcatcacc cacacagaca
     1801 gataccccca cggggtggtg ctgatcgacg acctgaccgc catggacccc agcctcgtgt
     1861 ctgctctggc ccccctgttt ggcagagccg cccacttccc cgagcagctg accagaccta
     1921 gcaccgccac agccgctggc gccagacacg ccgctgccaa gagcctgcag accatgcacg
     1981 agagcaccaa gggcctgggc cctgatggcg gaagaggctc tgcagccaag aagggcgagg
     2041 acgagaagtg ggaggctgcc gctggaacag gcggagccca gccaagcaca agcgagatgg
     2101 ccagctccag cctgtcctcc ggccagccac ctcctccact gagccagctg atcatcttcg
     2161 tgaccaccga cttcggccgg cagggccgga ccgtgggaaa gagcagaacc gagatcgagg
     2221 ccatgatcca gcacgacttc gccgacctgt acggcaccct gctgcccgcc tacacccgga
     2281 ccttcgtgta ctaccccttc accagcgcca tggctgagga tgtcgtgcgg agcgccgtga
     2341 ccgatctgcc ttgtgccctg ggcgagagac tgatcgcctc cagcaccatc tccgatgagg
     2401 ccgtgtcctt cctggtgcag cagcacagac agctgtggtc cggcaaagag aacggccacg
     2461 ccctgcggcg gctggtggaa gatgagatcg tgtctcagct ggtggtgtac tgggagacac
     2521 acgcccagca gaaacacttc gagaggctgc gggtgcactt tgagctggac gaggccagca
     2581 tgagagtgct gctgaggacc ccccacacca gccggacaac cgcctctcag cctcctgccg
     2641 ccctgtccag cgtggaagcc agagcctatg gccacggcgg agctgacgat ggcgacggcg
     2701 aggaaagaag cgatctgggc gcgcccggca gcctgcacca catcctggac gcccagaaga
     2761 tgctgtggaa ccacagggac aggaacctgc ctcctctggc ccctctgggc ccccaccacc
     2821 atcaccacca ttgagtctag acacgtgatt aattaaggat cccccgacct cgacctctgg
     2881 ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact
     2941 cggaaggaca tatgggaggg caaatcattt ggtcgagatc cctcggagat ctctagctag
     3001 aggatcgatc cccgccccgg acgaactaaa cctgactacg acatctctgc cccttcttcg
     3061 cggggcagtg catgtaatcc cttcagttgg ttggtacaac ttgccaactg ggccctgttc
     3121 cacatgtgac acgggggggg accaaacaca aaggggttct ctgactgtag ttgacatcct
     3181 tataaatgga tgtgcacatt tgccaacact gagtggcttt catcctggag cagactttgc
     3241 agtctgtgga ctgcaacaca acattgcctt tatgtgtaac tcttggctga agctcttaca
     3301 ccaatgctgg gggacatgta cctcccaggg gcccaggaag actacgggag gctacaccaa
     3361 cgtcaatcag aggggcctgt gtagctaccg ataagcggac cctcaagagg gcattagcaa
     3421 tagtgtttat aaggccccct tgttaaccct aaacgggtag catatgcttc ccgggtagta
     3481 gtatatacta tccagactaa ccctaattca atagcatatg ttacccaacg ggaagcatat
     3541 gctatcgaat tagggttagt aaaagggtcc taaggaacag cgatatctcc caccccatga
     3601 gctgtcacgg ttttatttac atggggtcag gattccacga gggtagtgaa ccattttagt
     3661 cacaagggca gtggctgaag atcaaggagc gggcagtgaa ctctcctgaa tcttcgcctg
     3721 cttcttcatt ctccttcgtt tagctaatag aataactgct gagttgtgaa cagtaaggtg
     3781 tatgtgaggt gctcgaaaac aaggtttcag gtgacgcccc cagaataaaa tttggacggg
     3841 gggttcagtg gtggcattgt gctatgacac caatataacc ctcacaaacc ccttgggcaa
     3901 taaatactag tgtaggaatg aaacattctg aatatcttta acaatagaaa tccatggggt
     3961 ggggacaagc cgtaaagact ggatgtccat ctcacacgaa tttatggcta tgggcaacac
     4021 ataatcctag tgcaatatga tactggggtt attaagatgt gtcccaggca gggaccaaga
     4081 caggtgaacc atgttgttac actctatttg taacaagggg aaagagagtg gacgccgaca
     4141 gcagcggact ccactggttg tctctaacac ccccgaaaat taaacggggc tccacgccaa
     4201 tggggcccat aaacaaagac aagtggccac tctttttttt gaaattgtgg agtgggggca
     4261 cgcgtcagcc cccacacgcc gccctgcggt tttggactgt aaaataaggg tgtaataact
     4321 tggctgattg taaccccgct aaccactgcg gtcaaaccac ttgcccacaa aaccactaat
     4381 ggcaccccgg ggaatacctg cataagtagg tgggcgggcc aagatagggg cgcgattgct
     4441 gcgatctgga ggacaaatta cacacacttg cgcctgagcg ccaagcacag ggttgttggt
     4501 cctcatattc acgaggtcgc tgagagcacg gtgggctaat gttgccatgg gtagcatata
     4561 ctacccaaat atctggatag catatgctat cctaatctat atctgggtag cataggctat
     4621 cctaatctat atctgggtag catatgctat cctaatctat atctgggtag tatatgctat
     4681 cctaatttat atctgggtag cataggctat cctaatctat atctgggtag catatgctat
     4741 cctaatctat atctgggtag tatatgctat cctaatctgt atccgggtag catatgctat
     4801 cctaatagag attagggtag tatatgctat cctaatttat atctgggtag catatactac
     4861 ccaaatatct ggatagcata tgctatccta atctatatct gggtagcata tgctatccta
     4921 atctatatct gggtagcata ggctatccta atctatatct gggtagcata tgctatccta
     4981 atctatatct gggtagtata tgctatccta atttatatct gggtagcata ggctatccta
     5041 atctatatct gggtagcata tgctatccta atctatatct gggtagtata tgctatccta
     5101 atctgtatcc gggtagcata tgctatcctc atgataagct gtcaaacatg agaattttct
     5161 tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     5221 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     5281 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     5341 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     5401 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     5461 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     5521 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     5581 ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc
     5641 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     5701 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     5761 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     5821 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     5881 aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta
     5941 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     6001 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     6061 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     6121 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     6181 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     6241 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     6301 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     6361 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6421 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6481 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6541 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6601 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6661 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6721 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6781 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6841 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6901 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6961 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     7021 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     7081 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     7141 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     7201 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     7261 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     7321 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     7381 tatgaccatg attacgccaa gctctagcta gaggtcgacc aattctcatg tttgacagct
     7441 tatcatcgca gatccgggca acgttgttgc cattgctgca ggcgcagaac tggtaggtat
     7501 ggaagatcta tacattgaat caatattggc aattagccat attagtcatt ggttatatag
     7561 cataaatcaa tattggctat tggccattgc atacgttgta tctatatcat aatatgtaca
     7621 tttatattgg ctcatgtcca atatgaccgc catgtt
//
