LOCUS       sequence-262330-        7470 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 7470)
  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 7470)
  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: D37_Ld_361830
FEATURES             Location/Qualifiers
     source          1..7470
                     /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             2634..2651
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2703..2758
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2757..2776)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4994..5012)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5080..5184
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5185..5253,5254..6045)
                     /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(5403..5422)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6216..6804
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6705..6724
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6958..6975
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7092..7113
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7128..7133,7134..7151,7152..7158)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7166..7182
                     /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     7171..7193
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7190..7206
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7190..7206
                     /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 ccctgtgccc cagcaatggc aacacccaca ccgtgtacgt gaccagcaac atgggcgaca
     1261 acatctggtc cttcgacacc ctgggcaact acctgggccc cgtgctgaac aaggccagct
     1321 tcccacccgg cgtgcgggtg gacaagctgc ggagcatgag attcgggccc cagggccacc
     1381 tgtatgtgtc cagcgccaga ggcagctaca gccgggtgtt cgccgtgtcc ggcaacggcc
     1441 tgctgaacgg caccatggaa gagaactgca cccgggacta cctgttcacc gtgatcgagc
     1501 aggacaaaga gaaccccttc ctggaccacc cctacgacat caccttccac cccgccaccg
     1561 aggatatgta cgtgtccaac cagaacagcg tgaccgtgac ccggtacacc agataccgga
     1621 ccgccgacgt gacccccaaa gccggcgctg gcggaagcca gcacgccttc tatcctgcct
     1681 ggaagcccgc caagaacgtg aaggccgccg tggaagccga cgaggaaacc gtgtctgccg
     1741 aagagacagt ggtggccccc aagaatatcc ccgacagcag cggcctgttc gccagcagct
     1801 ggtccagctc ctacagcatg tcctccgtgc ggggcctggc catcagccct ctgctgccta
     1861 gagccctggt ggaaggcgga gcccccagcg gagtgtttgc cgtgaccaac gagagcacca
     1921 tgtactacct gctcgtgtgc gacgtgtcct ccaataccgt gcacgtgttc gaccccgaga
     1981 gcggcgagag actgttcggc ctgaacgtgc ccagccccat ccaggtcatg ttccccaacc
     2041 ggtactacat cagcgacagc tccagcctga gcgtgtccgc cgctggcgtg cacacccaga
     2101 gattcgacgt gccctacatc tacgtgacat ccaaagagga cggcatggcc tacatggtcc
     2161 gattcagcag ccggcagggc aataccaaca acaacggcat gcagggggga gccttcggcg
     2221 aggacttcat ccgggcccac cggctgtaca ccatcaacca ccctaccccc ctgcacgccg
     2281 ccagcggcat ctacgagaat agcgccaggg acgtgctgct gatcgccgac cggaacggca
     2341 gacggatcta cagctacgcc agccctttcc tgaccgacta caccagcggc tacggcccct
     2401 ccccctttct gggcttcttc gtgaagcacc tccccgacca gcccgagttc atcatgagca
     2461 ccctgctgga acaccaggaa aacatcccat tctgctacga actcaacaag gacgggaagt
     2521 tcagatacgt ggccctgggc gcgcccggca gcctgcacca catcctggac gcccagaaga
     2581 tgctgtggaa ccacagggac aggaacctgc ctcctctggc ccctctgggc ccccaccacc
     2641 atcaccacca ttgagtctag acacgtgatt aattaaggat cccccgacct cgacctctgg
     2701 ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact
     2761 cggaaggaca tatgggaggg caaatcattt ggtcgagatc cctcggagat ctctagctag
     2821 aggatcgatc cccgccccgg acgaactaaa cctgactacg acatctctgc cccttcttcg
     2881 cggggcagtg catgtaatcc cttcagttgg ttggtacaac ttgccaactg ggccctgttc
     2941 cacatgtgac acgggggggg accaaacaca aaggggttct ctgactgtag ttgacatcct
     3001 tataaatgga tgtgcacatt tgccaacact gagtggcttt catcctggag cagactttgc
     3061 agtctgtgga ctgcaacaca acattgcctt tatgtgtaac tcttggctga agctcttaca
     3121 ccaatgctgg gggacatgta cctcccaggg gcccaggaag actacgggag gctacaccaa
     3181 cgtcaatcag aggggcctgt gtagctaccg ataagcggac cctcaagagg gcattagcaa
     3241 tagtgtttat aaggccccct tgttaaccct aaacgggtag catatgcttc ccgggtagta
     3301 gtatatacta tccagactaa ccctaattca atagcatatg ttacccaacg ggaagcatat
     3361 gctatcgaat tagggttagt aaaagggtcc taaggaacag cgatatctcc caccccatga
     3421 gctgtcacgg ttttatttac atggggtcag gattccacga gggtagtgaa ccattttagt
     3481 cacaagggca gtggctgaag atcaaggagc gggcagtgaa ctctcctgaa tcttcgcctg
     3541 cttcttcatt ctccttcgtt tagctaatag aataactgct gagttgtgaa cagtaaggtg
     3601 tatgtgaggt gctcgaaaac aaggtttcag gtgacgcccc cagaataaaa tttggacggg
     3661 gggttcagtg gtggcattgt gctatgacac caatataacc ctcacaaacc ccttgggcaa
     3721 taaatactag tgtaggaatg aaacattctg aatatcttta acaatagaaa tccatggggt
     3781 ggggacaagc cgtaaagact ggatgtccat ctcacacgaa tttatggcta tgggcaacac
     3841 ataatcctag tgcaatatga tactggggtt attaagatgt gtcccaggca gggaccaaga
     3901 caggtgaacc atgttgttac actctatttg taacaagggg aaagagagtg gacgccgaca
     3961 gcagcggact ccactggttg tctctaacac ccccgaaaat taaacggggc tccacgccaa
     4021 tggggcccat aaacaaagac aagtggccac tctttttttt gaaattgtgg agtgggggca
     4081 cgcgtcagcc cccacacgcc gccctgcggt tttggactgt aaaataaggg tgtaataact
     4141 tggctgattg taaccccgct aaccactgcg gtcaaaccac ttgcccacaa aaccactaat
     4201 ggcaccccgg ggaatacctg cataagtagg tgggcgggcc aagatagggg cgcgattgct
     4261 gcgatctgga ggacaaatta cacacacttg cgcctgagcg ccaagcacag ggttgttggt
     4321 cctcatattc acgaggtcgc tgagagcacg gtgggctaat gttgccatgg gtagcatata
     4381 ctacccaaat atctggatag catatgctat cctaatctat atctgggtag cataggctat
     4441 cctaatctat atctgggtag catatgctat cctaatctat atctgggtag tatatgctat
     4501 cctaatttat atctgggtag cataggctat cctaatctat atctgggtag catatgctat
     4561 cctaatctat atctgggtag tatatgctat cctaatctgt atccgggtag catatgctat
     4621 cctaatagag attagggtag tatatgctat cctaatttat atctgggtag catatactac
     4681 ccaaatatct ggatagcata tgctatccta atctatatct gggtagcata tgctatccta
     4741 atctatatct gggtagcata ggctatccta atctatatct gggtagcata tgctatccta
     4801 atctatatct gggtagtata tgctatccta atttatatct gggtagcata ggctatccta
     4861 atctatatct gggtagcata tgctatccta atctatatct gggtagtata tgctatccta
     4921 atctgtatcc gggtagcata tgctatcctc atgataagct gtcaaacatg agaattttct
     4981 tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     5041 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     5101 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     5161 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     5221 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     5281 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     5341 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     5401 ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc
     5461 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     5521 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     5581 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     5641 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     5701 aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta
     5761 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     5821 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     5881 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     5941 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     6001 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     6061 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     6121 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     6181 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6241 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6301 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6361 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6421 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6481 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6541 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6601 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6661 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6721 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6781 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6841 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     6901 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     6961 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     7021 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     7081 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     7141 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     7201 tatgaccatg attacgccaa gctctagcta gaggtcgacc aattctcatg tttgacagct
     7261 tatcatcgca gatccgggca acgttgttgc cattgctgca ggcgcagaac tggtaggtat
     7321 ggaagatcta tacattgaat caatattggc aattagccat attagtcatt ggttatatag
     7381 cataaatcaa tattggctat tggccattgc atacgttgta tctatatcat aatatgtaca
     7441 tttatattgg ctcatgtcca atatgaccgc
//
