LOCUS       sequence-262343-        8310 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 8310)
  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 8310)
  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: D51_Ld_310120
FEATURES             Location/Qualifiers
     source          1..8310
                     /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             3474..3491
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3543..3598
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3597..3616)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5834..5852)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5920..6024
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6025..6093,6094..6885)
                     /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(6243..6262)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7056..7644
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7545..7564
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7798..7815
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7932..7953
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7968..7973,7974..7991,7992..7998)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8006..8022
                     /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     8011..8033
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8030..8046
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8030..8046
                     /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 gcggccgccc
     1201 aggatgtgga acagctggtg cccaacagca ccagccagtg ctaccccagc atcgacctgg
     1261 ggtggtccgc ccacgtggga agcagcgtgt acgccacccc cagaatcgtg gacctggccc
     1321 acgacggcca caaagaggtg ctgatcccta ccttcagcca gttcctggaa gccctggacg
     1381 gccccaccgg cgaggatatc cccggcttcc cattcatgca cccccggatg aagacctacg
     1441 ccagccccct gcccgtggac ctgaatggcg acggacaggt ggactggctg gtggccctgt
     1501 acagcggcga gctggtggtg ttcggccagg acggcaaggt gcagggctac ctccaggtgc
     1561 cccccctggc catgaagcgg aactgggtca aggacaacct gcccccccag caggccgcca
     1621 cagtcagacc tggcaccccc ctgaaaaccg gcaccgtgct ggaagatatc ctgaaggcca
     1681 gacggatgga cgacctgacc ctgaccagca ccgaggtgct ggaccagcgg cggagaagca
     1741 gcatcgtgca gagacaccac gcccccaccg ccgctgccgt gaagtccgtg gaatacgaca
     1801 cccaccccaa gggcgccgag ctgagagcca ccgaggacaa gcccgaccag tggaacggcg
     1861 acgacgccct ggacatcgac gacgacgaga tggaactgcc ctgggagaag gccgaggtgg
     1921 aagtgggccc caacagccac ctgagccctg aggccagagc cagcctggac ctgctgttcc
     1981 accccgagct gtaccagacc gccaagagag ccacaagcgc cgacgaggcc accttcgagg
     2041 gccggaatct gcggcaggcc acctctatcg ccttcggcga ggacgagatc aaggtggacg
     2101 cccacatcct gagcaccccc gtggtcaccg acgtggacca gaacggcgat ctggacgtga
     2161 tcctgcacgt gtcctacttt ttcggagagg acgtgatcgc cgagcagccc gagagattcg
     2221 ctggcggcgt ggacccccac gatttcgtgg ccacagccgt cgtgtgcctg aacctggtca
     2281 ccggcgagac aagatgggcc agactgctgc acgccagcac caagaacagc accaacgccg
     2341 cctacgccct gtccaccccc ctggtcatca acgccgatga ggaccggtcc atggaagtgt
     2401 tcgtgtccac cagcctgggc ttcgtgttcg gcttcaccgc cgacggcaac accctggatg
     2461 gctggcctgt gctgctgggc cctctgagcg ccagccctac cgccgaagat atcacaggcg
     2521 acggcaccct ggatatctgc atcggcgaca ccgagggcaa agtgcggtgc tacaacgcca
     2581 gcggcagcgt gaccctggaa aaggatgtga tcggcggcgt ggccgaccgg ctgaccttcg
     2641 gagatgtgga tggcgacggc gtggtggaca tcgtgtttgg caccaccagc ggcctgatct
     2701 acgccctgag aggcaccgat ggcgccgtgc tgcccagctt ccctatcgtg accggcggct
     2761 ctatcgtggc ccctgccctg ctgctgaacc tgaacggcat gctgtactcc aacgacctgg
     2821 atatcgtggt gcccagccac aacggccggg tgtacctgct gagaggcagc agcggctgcg
     2881 tgcagaccat cgacatcgat gagaagtcca gcagcatggt gctggccgac gacgtgctgg
     2941 gcaacggcca gatggacctg gtggtcacca ccatcaaggg cggcgtgtac gtgttccaga
     3001 ccagcacccc ctacaccgcc atgaacgcct ggcccagcaa gaccaagggc gtgaacggct
     3061 tcagcgcctc cggcaactac atcggaatcg ccatcagccc caacttccgg accttccggg
     3121 acatccaggg cgacaccttc aacgtggaat tcaccatcta cgacagcaga cccgtgtccc
     3181 ccaagcggag ctacaccgtg gaaatcaccg tgggcacccg gatccggctg tacagagagc
     3241 ggttccacaa gcccggcacc cacatcgtga aagtgcgggc acccctggaa cggatgtaca
     3301 gcagcctgaa cgtggccctg accctgccca acggccagtc cttcaccgac agcatcagcc
     3361 tgagcttcaa catgcatggc gcgcccggca gcctgcacca catcctggac gcccagaaga
     3421 tgctgtggaa ccacagggac aggaacctgc ctcctctggc ccctctgggc ccccaccacc
     3481 atcaccacca ttgagtctag acacgtgatt aattaaggat cccccgacct cgacctctgg
     3541 ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact
     3601 cggaaggaca tatgggaggg caaatcattt ggtcgagatc cctcggagat ctctagctag
     3661 aggatcgatc cccgccccgg acgaactaaa cctgactacg acatctctgc cccttcttcg
     3721 cggggcagtg catgtaatcc cttcagttgg ttggtacaac ttgccaactg ggccctgttc
     3781 cacatgtgac acgggggggg accaaacaca aaggggttct ctgactgtag ttgacatcct
     3841 tataaatgga tgtgcacatt tgccaacact gagtggcttt catcctggag cagactttgc
     3901 agtctgtgga ctgcaacaca acattgcctt tatgtgtaac tcttggctga agctcttaca
     3961 ccaatgctgg gggacatgta cctcccaggg gcccaggaag actacgggag gctacaccaa
     4021 cgtcaatcag aggggcctgt gtagctaccg ataagcggac cctcaagagg gcattagcaa
     4081 tagtgtttat aaggccccct tgttaaccct aaacgggtag catatgcttc ccgggtagta
     4141 gtatatacta tccagactaa ccctaattca atagcatatg ttacccaacg ggaagcatat
     4201 gctatcgaat tagggttagt aaaagggtcc taaggaacag cgatatctcc caccccatga
     4261 gctgtcacgg ttttatttac atggggtcag gattccacga gggtagtgaa ccattttagt
     4321 cacaagggca gtggctgaag atcaaggagc gggcagtgaa ctctcctgaa tcttcgcctg
     4381 cttcttcatt ctccttcgtt tagctaatag aataactgct gagttgtgaa cagtaaggtg
     4441 tatgtgaggt gctcgaaaac aaggtttcag gtgacgcccc cagaataaaa tttggacggg
     4501 gggttcagtg gtggcattgt gctatgacac caatataacc ctcacaaacc ccttgggcaa
     4561 taaatactag tgtaggaatg aaacattctg aatatcttta acaatagaaa tccatggggt
     4621 ggggacaagc cgtaaagact ggatgtccat ctcacacgaa tttatggcta tgggcaacac
     4681 ataatcctag tgcaatatga tactggggtt attaagatgt gtcccaggca gggaccaaga
     4741 caggtgaacc atgttgttac actctatttg taacaagggg aaagagagtg gacgccgaca
     4801 gcagcggact ccactggttg tctctaacac ccccgaaaat taaacggggc tccacgccaa
     4861 tggggcccat aaacaaagac aagtggccac tctttttttt gaaattgtgg agtgggggca
     4921 cgcgtcagcc cccacacgcc gccctgcggt tttggactgt aaaataaggg tgtaataact
     4981 tggctgattg taaccccgct aaccactgcg gtcaaaccac ttgcccacaa aaccactaat
     5041 ggcaccccgg ggaatacctg cataagtagg tgggcgggcc aagatagggg cgcgattgct
     5101 gcgatctgga ggacaaatta cacacacttg cgcctgagcg ccaagcacag ggttgttggt
     5161 cctcatattc acgaggtcgc tgagagcacg gtgggctaat gttgccatgg gtagcatata
     5221 ctacccaaat atctggatag catatgctat cctaatctat atctgggtag cataggctat
     5281 cctaatctat atctgggtag catatgctat cctaatctat atctgggtag tatatgctat
     5341 cctaatttat atctgggtag cataggctat cctaatctat atctgggtag catatgctat
     5401 cctaatctat atctgggtag tatatgctat cctaatctgt atccgggtag catatgctat
     5461 cctaatagag attagggtag tatatgctat cctaatttat atctgggtag catatactac
     5521 ccaaatatct ggatagcata tgctatccta atctatatct gggtagcata tgctatccta
     5581 atctatatct gggtagcata ggctatccta atctatatct gggtagcata tgctatccta
     5641 atctatatct gggtagtata tgctatccta atttatatct gggtagcata ggctatccta
     5701 atctatatct gggtagcata tgctatccta atctatatct gggtagtata tgctatccta
     5761 atctgtatcc gggtagcata tgctatcctc atgataagct gtcaaacatg agaattttct
     5821 tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     5881 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     5941 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     6001 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     6061 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     6121 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     6181 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     6241 ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc
     6301 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     6361 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     6421 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     6481 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     6541 aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta
     6601 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     6661 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     6721 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     6781 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     6841 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     6901 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     6961 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     7021 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     7081 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     7141 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     7201 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     7261 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     7321 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     7381 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     7441 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     7501 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     7561 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     7621 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     7681 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     7741 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     7801 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     7861 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     7921 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     7981 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     8041 tatgaccatg attacgccaa gctctagcta gaggtcgacc aattctcatg tttgacagct
     8101 tatcatcgca gatccgggca acgttgttgc cattgctgca ggcgcagaac tggtaggtat
     8161 ggaagatcta tacattgaat caatattggc aattagccat attagtcatt ggttatatag
     8221 cataaatcaa tattggctat tggccattgc atacgttgta tctatatcat aatatgtaca
     8281 tttatattgg ctcatgtcca atatgaccgc
//
