LOCUS       sequence-262367-        6891 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 6891)
  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 6891)
  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: D75_Ld_302670
FEATURES             Location/Qualifiers
     source          1..6891
                     /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             2055..2072
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2124..2179
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2178..2197)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4415..4433)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4501..4605
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4606..4674,4675..5466)
                     /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(4824..4843)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5637..6225
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6126..6145
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6379..6396
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6513..6534
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6549..6554,6555..6572,6573..6579)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6587..6603
                     /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     6592..6614
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6611..6627
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6611..6627
                     /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 gcggccgccg
     1201 aagtagttgt cgcccccttg tgtgcagaga aacatggttt cgccggagaa tttaacgatc
     1261 attacaccat accagtagaa caggcgccgg ctgctggtga ggccatcgtg ctcatcgccc
     1321 gggcctttcc actgaatttg ggaaatatgc tcgctaggtg gctgtatatc aatgccacct
     1381 accggatcag cgaaggcgca caacttaccg aggtctccga caataacggt atccttgagc
     1441 tggccaacgt ggctgccgga accgatatcg aggttcgagt aatgagagtg cgtacagacg
     1501 ggccgacacc atttaggttc ttcagcttct tcgccaactc tcctgtgtgc catgttgcag
     1561 tggcaccaga taactctttc attgctcccg ttccgaggcg tctgagagcg accccacagt
     1621 tgcctttgac gatgtacttt aaggcagtgc tgcccccgaa tgttaacgcc cttatcatcc
     1681 gagtaagcgc agacggacgc accgatgaga aatttagaag cggcgataag gtgtatgctt
     1741 ctggcgacct ggtccgcgca gcatcaggtg gcgcagtgct gtttgcatat actttcccta
     1801 tggaaaacgc tgccactccc tactacttca cccaagtcag tgtgagctat ggcgagtggg
     1861 aaggtggcga cgcggccaag ccgtctgcat ctaacacgag ttcatctggc ggtgactcta
     1921 aagacacgat gcaacccaca cgaagtacgc ccgtgcttgg cgcgcccggc agcctgcacc
     1981 acatcctgga cgcccagaag atgctgtgga accacaggga caggaacctg cctcctctgg
     2041 cccctctggg cccccaccac catcaccacc attgagtcta gacacgtgat taattaagga
     2101 tcccccgacc tcgacctctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg
     2161 gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt tggtcgagat
     2221 ccctcggaga tctctagcta gaggatcgat ccccgccccg gacgaactaa acctgactac
     2281 gacatctctg ccccttcttc gcggggcagt gcatgtaatc ccttcagttg gttggtacaa
     2341 cttgccaact gggccctgtt ccacatgtga cacggggggg gaccaaacac aaaggggttc
     2401 tctgactgta gttgacatcc ttataaatgg atgtgcacat ttgccaacac tgagtggctt
     2461 tcatcctgga gcagactttg cagtctgtgg actgcaacac aacattgcct ttatgtgtaa
     2521 ctcttggctg aagctcttac accaatgctg ggggacatgt acctcccagg ggcccaggaa
     2581 gactacggga ggctacacca acgtcaatca gaggggcctg tgtagctacc gataagcgga
     2641 ccctcaagag ggcattagca atagtgttta taaggccccc ttgttaaccc taaacgggta
     2701 gcatatgctt cccgggtagt agtatatact atccagacta accctaattc aatagcatat
     2761 gttacccaac gggaagcata tgctatcgaa ttagggttag taaaagggtc ctaaggaaca
     2821 gcgatatctc ccaccccatg agctgtcacg gttttattta catggggtca ggattccacg
     2881 agggtagtga accattttag tcacaagggc agtggctgaa gatcaaggag cgggcagtga
     2941 actctcctga atcttcgcct gcttcttcat tctccttcgt ttagctaata gaataactgc
     3001 tgagttgtga acagtaaggt gtatgtgagg tgctcgaaaa caaggtttca ggtgacgccc
     3061 ccagaataaa atttggacgg ggggttcagt ggtggcattg tgctatgaca ccaatataac
     3121 cctcacaaac cccttgggca ataaatacta gtgtaggaat gaaacattct gaatatcttt
     3181 aacaatagaa atccatgggg tggggacaag ccgtaaagac tggatgtcca tctcacacga
     3241 atttatggct atgggcaaca cataatccta gtgcaatatg atactggggt tattaagatg
     3301 tgtcccaggc agggaccaag acaggtgaac catgttgtta cactctattt gtaacaaggg
     3361 gaaagagagt ggacgccgac agcagcggac tccactggtt gtctctaaca cccccgaaaa
     3421 ttaaacgggg ctccacgcca atggggccca taaacaaaga caagtggcca ctcttttttt
     3481 tgaaattgtg gagtgggggc acgcgtcagc ccccacacgc cgccctgcgg ttttggactg
     3541 taaaataagg gtgtaataac ttggctgatt gtaaccccgc taaccactgc ggtcaaacca
     3601 cttgcccaca aaaccactaa tggcaccccg gggaatacct gcataagtag gtgggcgggc
     3661 caagataggg gcgcgattgc tgcgatctgg aggacaaatt acacacactt gcgcctgagc
     3721 gccaagcaca gggttgttgg tcctcatatt cacgaggtcg ctgagagcac ggtgggctaa
     3781 tgttgccatg ggtagcatat actacccaaa tatctggata gcatatgcta tcctaatcta
     3841 tatctgggta gcataggcta tcctaatcta tatctgggta gcatatgcta tcctaatcta
     3901 tatctgggta gtatatgcta tcctaattta tatctgggta gcataggcta tcctaatcta
     3961 tatctgggta gcatatgcta tcctaatcta tatctgggta gtatatgcta tcctaatctg
     4021 tatccgggta gcatatgcta tcctaataga gattagggta gtatatgcta tcctaattta
     4081 tatctgggta gcatatacta cccaaatatc tggatagcat atgctatcct aatctatatc
     4141 tgggtagcat atgctatcct aatctatatc tgggtagcat aggctatcct aatctatatc
     4201 tgggtagcat atgctatcct aatctatatc tgggtagtat atgctatcct aatttatatc
     4261 tgggtagcat aggctatcct aatctatatc tgggtagcat atgctatcct aatctatatc
     4321 tgggtagtat atgctatcct aatctgtatc cgggtagcat atgctatcct catgataagc
     4381 tgtcaaacat gagaattttc ttgaagacga aagggcctcg tgatacgcct atttttatag
     4441 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     4501 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     4561 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     4621 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     4681 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     4741 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     4801 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg
     4861 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     4921 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     4981 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     5041 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     5101 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca
     5161 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     5221 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     5281 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     5341 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     5401 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     5461 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     5521 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     5581 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5641 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5701 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     5761 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     5821 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     5881 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     5941 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6001 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6061 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6121 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6181 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6241 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     6301 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     6361 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     6421 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     6481 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     6541 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     6601 caatttcaca caggaaacag ctatgaccat gattacgcca agctctagct agaggtcgac
     6661 caattctcat gtttgacagc ttatcatcgc agatccgggc aacgttgttg ccattgctgc
     6721 aggcgcagaa ctggtaggta tggaagatct atacattgaa tcaatattgg caattagcca
     6781 tattagtcat tggttatata gcataaatca atattggcta ttggccattg catacgttgt
     6841 atctatatca taatatgtac atttatattg gctcatgtcc aatatgaccg c
//
