LOCUS       sequence-275114-        6376 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 6376)
  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 6376)
  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: D86_Ld_260650
FEATURES             Location/Qualifiers
     source          1..6376
                     /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             1710..1727
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1779..1834
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1833..1852)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3894..3912)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3980..4084
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4085..4153,4154..4945)
                     /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(4303..4322)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5116..5704
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5605..5624
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5858..5875
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5992..6013
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6028..6033,6034..6051,6052..6058)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6066..6082
                     /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     6071..6093
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6090..6106
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6090..6106
                     /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 gccgaagaag
     1201 gaacggttac atactacggt cagctgagat tgccgccaac gtacctgcgt catccagatt
     1261 gctttgaggc cctcaacgat atccagcccg gcagcgtgct gttgtataac ggccagcaca
     1321 gatttgtcgt gccaactgcc agggacggct ccttctccgt ctataagctg ccctatggga
     1381 catacattct gcaggccgag tatcattact ttatgttccc caccgtgcgc gtcgaggtta
     1441 tgtaccgcga taccggcgac gaccaaaagg agacttttat acgtacatca gctaacgact
     1501 atcccgtgcg gcatctcgag gggtctggac tggacgagga atcaccagcc gtgattccct
     1561 ttagcggtta ccataactac tatatccccc gccaacagat ggacattgtt tccggcgcgc
     1621 ccggcagcct gcaccacatc ctggacgccc agaagatgct gtggaaccac agggacagga
     1681 acctgcctcc tctggcccct ctgggccccc accaccatca ccaccattga gtctagacac
     1741 gtgattaatt aaggatcccc cgacctcgac ctctggctaa taaaggaaat ttattttcat
     1801 tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa
     1861 tcatttggtc gagatccctc ggagatctct agctagagga tcgatccccg ccccggacga
     1921 actaaacctg actacgacat ctctgcccct tcttcgcggg gcagtgcatg taatcccttc
     1981 agttggttgg tacaacttgc caactgggcc ctgttccaca tgtgacacgg ggggggacca
     2041 aacacaaagg ggttctctga ctgtagttga catccttata aatggatgtg cacatttgcc
     2101 aacactgagt ggctttcatc ctggagcaga ctttgcagtc tgtggactgc aacacaacat
     2161 tgcctttatg tgtaactctt ggctgaagct cttacaccaa tgctggggga catgtacctc
     2221 ccaggggccc aggaagacta cgggaggcta caccaacgtc aatcagaggg gcctgtgtag
     2281 ctaccgataa gcggaccctc aagagggcat tagcaatagt gtttataagg cccccttgtt
     2341 aaccctaaac gggtagcata tgcttcccgg gtagtagtat atactatcca gactaaccct
     2401 aattcaatag catatgttac ccaacgggaa gcatatgcta tcgaattagg gttagtaaaa
     2461 gggtcctaag gaacagcgat atctcccacc ccatgagctg tcacggtttt atttacatgg
     2521 ggtcaggatt ccacgagggt agtgaaccat tttagtcaca agggcagtgg ctgaagatca
     2581 aggagcgggc agtgaactct cctgaatctt cgcctgcttc ttcattctcc ttcgtttagc
     2641 taatagaata actgctgagt tgtgaacagt aaggtgtatg tgaggtgctc gaaaacaagg
     2701 tttcaggtga cgcccccaga ataaaatttg gacggggggt tcagtggtgg cattgtgcta
     2761 tgacaccaat ataaccctca caaacccctt gggcaataaa tactagtgta ggaatgaaac
     2821 attctgaata tctttaacaa tagaaatcca tggggtgggg acaagccgta aagactggat
     2881 gtccatctca cacgaattta tggctatggg caacacataa tcctagtgca atatgatact
     2941 ggggttatta agatgtgtcc caggcaggga ccaagacagg tgaaccatgt tgttacactc
     3001 tatttgtaac aaggggaaag agagtggacg ccgacagcag cggactccac tggttgtctc
     3061 taacaccccc gaaaattaaa cggggctcca cgccaatggg gcccataaac aaagacaagt
     3121 ggccactctt ttttttgaaa ttgtggagtg ggggcacgcg tcagccccca cacgccgccc
     3181 tgcggttttg gactgtaaaa taagggtgta ataacttggc tgattgtaac cccgctaacc
     3241 actgcggtca aaccacttgc ccacaaaacc actaatggca ccccggggaa tacctgcata
     3301 agtaggtggg cgggccaaga taggggcgcg attgctgcga tctggaggac aaattacaca
     3361 cacttgcgcc tgagcgccaa gcacagggtt gttggtcctc atattcacga ggtcgctgag
     3421 agcacggtgg gctaatgttg ccatgggtag catatactac ccaaatatct ggatagcata
     3481 tgctatccta atctatatct gggtagcata ggctatccta atctatatct gggtagcata
     3541 tgctatccta atctatatct gggtagtata tgctatccta atttatatct gggtagcata
     3601 ggctatccta atctatatct gggtagcata tgctatccta atctatatct gggtagtata
     3661 tgctatccta atctgtatcc gggtagcata tgctatccta atagagatta gggtagtata
     3721 tgctatccta atttatatct gggtagcata ggctatccta atctatatct gggtagcata
     3781 tgctatccta atctatatct gggtagtata tgctatccta atctgtatcc gggtagcata
     3841 tgctatcctc atgataagct gtcaaacatg agaattttct tgaagacgaa agggcctcgt
     3901 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     3961 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     4021 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     4081 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     4141 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     4201 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     4261 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     4321 atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     4381 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     4441 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     4501 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     4561 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     4621 gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     4681 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     4741 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     4801 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     4861 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     4921 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     4981 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     5041 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5101 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5161 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5221 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     5281 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5341 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     5401 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     5461 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     5521 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     5581 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     5641 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     5701 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     5761 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     5821 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     5881 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     5941 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     6001 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     6061 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
     6121 gctctagcta gaggtcgacc aattctcatg tttgacagct tatcatcgca gatccgggca
     6181 acgttgttgc cattgctgca ggcgcagaac tggtaggtat ggaagatcta tacattgaat
     6241 caatattggc aattagccat attagtcatt ggttatatag cataaatcaa tattggctat
     6301 tggccattgc atacgttgta tctatatcat aatatgtaca tttatattgg ctcatgtcca
     6361 atatgaccgc catgtt
//
