LOCUS       sequence-262372-        9060 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 9060)
  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 9060)
  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: D80_Ld_220290
FEATURES             Location/Qualifiers
     source          1..9060
                     /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             4224..4241
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4293..4348
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4347..4366)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6584..6602)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6670..6774
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6775..6843,6844..7635)
                     /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(6993..7012)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7806..8394
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8295..8314
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8548..8565
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8682..8703
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8718..8723,8724..8741,8742..8748)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8756..8772
                     /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     8761..8783
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8780..8796
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8780..8796
                     /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 ggccttgtac gaacgccgac attatccact tcttctctcc ttgtgacaag cgaacgggaa
     1261 agctgaccgt gttgagctac cttagctcag atgcggattg cgagcgcggg aacctgtttg
     1321 ttcctcctcc caggacaaca gcatgctatg actgcaccga tggatatcgg ctggatcctc
     1381 tcacacttga atgcgccgtg tgtccagccg gtacttattc tagttatacg actaggtacg
     1441 tgaccttcga tcccttgcct gaggacatgc gcacttactg ttccccagaa ccatgtgcat
     1501 catggtccgc cgtggaagac acctatggca ctgttgtcag cagcggggtc cagcccactg
     1561 agaatcgccc tgttacccaa tggggcacca tcgacgaaag tgttgagtcc acgctggatt
     1621 tcgttgtcga tgttatagaa tccagcggct ccctggagtt cgattactcc atctcaagtg
     1681 aggagatgta cgacggactg gtgctccgac tcaattacag catcgtgcga aatcctgata
     1741 ccgaggtggg aaaatatcgg ttcttcgcaa ccggagtgca tagagactgg taccacgcta
     1801 gcctgagact cccctatggg accactcaaa tacaattcga ttacatgaag gatagtactg
     1861 ggcccccgta tggcgaagag ctcactggtg tgaatgacag agcactgttg cgcaatatcg
     1921 tagttaaagg tacaaggaag accgtcgcag aaatgcagtg cgtttcttgt ccacccggcc
     1981 actggacggc aagagagggg agcggtggtg tcggggccag tcaatgcgtc aggtgtccaa
     2041 gaaacacata caggggcgaa ggggacaccc agtgtctgca gtgtccaccc aacacatcaa
     2101 gcccagaagg ggctaataat tgtactcgca tgagagcttg taccggcggc gattacatcg
     2161 cacaatacgg ttcatgccgt agcgacaata ccagattgag atcatgggtg aaatacgaag
     2221 gcgctgattg tgcggaagtg gaaggttcac gccctacagc tacccgggtg gaatgcgcta
     2281 cctgtctgcc cggaatgcgg catgctgata gtagggctcg cggcgcttgc gtttcatgtc
     2341 ctgccgggga atatttgctc ggagacaggt gtgaggtgtg ccccttgggg acagccgcta
     2401 tacccaagct tgtgtacagt tccgggttcg atggtctggg cggcgacgtg atggccttgc
     2461 cggggatctc tctgagcaga atttactgca ctgggtccgc ctctcgctgt cgccgggaag
     2521 agggtaaggg gttcgaactc gtgggccttc catttgcaga tgaggacaac actctgatag
     2581 gcatccgcca ggagcacctc gcctcttcca catactctct ggctactctg agctacacct
     2641 ttcacgccga agaggacggc tgggcgaata ttacctttgc ttatgtcgac gctgaaggaa
     2701 agtatgtcca acgcatggac tggcgcacca ctctgaatgt gcgagtaatt gtggatgata
     2761 acgcagagta tcccgtgagg gcttttgcgt ctcacactgg ccgtgtggat tccgtgttct
     2821 acagcctgct tgtcccttat tatgtggata ccgcactgcc tgcttccgga tctgatagca
     2881 aattgagagg catgcgtcat cggctgtctt ggatcgtgga agaattggcc ttcacatcta
     2941 atccaatcag tgttgtgata gtcggactgg aagtcttcgg cgacgtctca ggtggggtgc
     3001 agacctgcca gccatgcagg gccgggttca gttgtgctag tagggcggct aagatgacac
     3061 cttgctcccc cggtactgtg cagccggcac tcagggctca tcgctgcgag ccttgtatag
     3121 ggaacacata cgcccctggc tatgggttta ctacctgcct gcagtgcccg gggaacactg
     3181 tcgccaatgc agaccatacc ggatgctcct tgggctgtgt atttgagcgc aacggcatgg
     3241 tctataactt cagtgagctt cgcggcgtgg tactcaacgc cagtgtcggc tacatgcctg
     3301 ccactaacgc ttcagtcagc aacatgactg tggaagaggc cgatgaagct ggtataccac
     3361 ggttttactt ttctccttgc gatccactgc ccgttagaga cgcgcgcgcg acgagcgaat
     3421 ccgactctgc gcttacccct atcacgagag atagactgtg cgtcccgtct gccagaggca
     3481 ggaacagctc cagctctgcc tacgtttgtc agcgcacctc tgcaacaaat gggcatcatt
     3541 ttggagacgc cgtggttcat atggaagtag gaagacacct ctatctggtg acgcaattgg
     3601 gctccccgta tattttctac ggtgttgata catccgcaga cgattgggtg tatgacgcta
     3661 agtccaatta catggcattc attgagctgg aatgtagcac atctggtggc gagcctaaat
     3721 ttggtacact gcagtacgtg ggtgaaggga atgaggacct ggtgctcagg tggcgttccg
     3781 cctatgcctg ccccctttgt acaaatgaat catacgagaa ggtggaaact tcatgcgatg
     3841 atcctacggc cacaaccaga agggtctact atacactgcg ccgcgactct atgaattgtg
     3901 tgggcgggta cgtcccaccc gaaccgataa ttgttaagtg tactgagtgc accctggatg
     3961 cttacactgt tagatggctg gaatgtgatc cctcaactca aactcaggaa ggggtgtatg
     4021 tgttgttgcc ggatcgggcg aactgcaccc ctggagctag tcctcttcca cccgtaaagc
     4081 caagaccgtg tgatgcttcc ggtggtactg ccggacgtgg caccagcggc gcgcccggca
     4141 gcctgcacca catcctggac gcccagaaga tgctgtggaa ccacagggac aggaacctgc
     4201 ctcctctggc ccctctgggc ccccaccacc atcaccacca ttgagtctag acacgtgatt
     4261 aattaaggat cccccgacct cgacctctgg ctaataaagg aaatttattt tcattgcaat
     4321 agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg caaatcattt
     4381 ggtcgagatc cctcggagat ctctagctag aggatcgatc cccgccccgg acgaactaaa
     4441 cctgactacg acatctctgc cccttcttcg cggggcagtg catgtaatcc cttcagttgg
     4501 ttggtacaac ttgccaactg ggccctgttc cacatgtgac acgggggggg accaaacaca
     4561 aaggggttct ctgactgtag ttgacatcct tataaatgga tgtgcacatt tgccaacact
     4621 gagtggcttt catcctggag cagactttgc agtctgtgga ctgcaacaca acattgcctt
     4681 tatgtgtaac tcttggctga agctcttaca ccaatgctgg gggacatgta cctcccaggg
     4741 gcccaggaag actacgggag gctacaccaa cgtcaatcag aggggcctgt gtagctaccg
     4801 ataagcggac cctcaagagg gcattagcaa tagtgtttat aaggccccct tgttaaccct
     4861 aaacgggtag catatgcttc ccgggtagta gtatatacta tccagactaa ccctaattca
     4921 atagcatatg ttacccaacg ggaagcatat gctatcgaat tagggttagt aaaagggtcc
     4981 taaggaacag cgatatctcc caccccatga gctgtcacgg ttttatttac atggggtcag
     5041 gattccacga gggtagtgaa ccattttagt cacaagggca gtggctgaag atcaaggagc
     5101 gggcagtgaa ctctcctgaa tcttcgcctg cttcttcatt ctccttcgtt tagctaatag
     5161 aataactgct gagttgtgaa cagtaaggtg tatgtgaggt gctcgaaaac aaggtttcag
     5221 gtgacgcccc cagaataaaa tttggacggg gggttcagtg gtggcattgt gctatgacac
     5281 caatataacc ctcacaaacc ccttgggcaa taaatactag tgtaggaatg aaacattctg
     5341 aatatcttta acaatagaaa tccatggggt ggggacaagc cgtaaagact ggatgtccat
     5401 ctcacacgaa tttatggcta tgggcaacac ataatcctag tgcaatatga tactggggtt
     5461 attaagatgt gtcccaggca gggaccaaga caggtgaacc atgttgttac actctatttg
     5521 taacaagggg aaagagagtg gacgccgaca gcagcggact ccactggttg tctctaacac
     5581 ccccgaaaat taaacggggc tccacgccaa tggggcccat aaacaaagac aagtggccac
     5641 tctttttttt gaaattgtgg agtgggggca cgcgtcagcc cccacacgcc gccctgcggt
     5701 tttggactgt aaaataaggg tgtaataact tggctgattg taaccccgct aaccactgcg
     5761 gtcaaaccac ttgcccacaa aaccactaat ggcaccccgg ggaatacctg cataagtagg
     5821 tgggcgggcc aagatagggg cgcgattgct gcgatctgga ggacaaatta cacacacttg
     5881 cgcctgagcg ccaagcacag ggttgttggt cctcatattc acgaggtcgc tgagagcacg
     5941 gtgggctaat gttgccatgg gtagcatata ctacccaaat atctggatag catatgctat
     6001 cctaatctat atctgggtag cataggctat cctaatctat atctgggtag catatgctat
     6061 cctaatctat atctgggtag tatatgctat cctaatttat atctgggtag cataggctat
     6121 cctaatctat atctgggtag catatgctat cctaatctat atctgggtag tatatgctat
     6181 cctaatctgt atccgggtag catatgctat cctaatagag attagggtag tatatgctat
     6241 cctaatttat atctgggtag catatactac ccaaatatct ggatagcata tgctatccta
     6301 atctatatct gggtagcata tgctatccta atctatatct gggtagcata ggctatccta
     6361 atctatatct gggtagcata tgctatccta atctatatct gggtagtata tgctatccta
     6421 atttatatct gggtagcata ggctatccta atctatatct gggtagcata tgctatccta
     6481 atctatatct gggtagtata tgctatccta atctgtatcc gggtagcata tgctatcctc
     6541 atgataagct gtcaaacatg agaattttct tgaagacgaa agggcctcgt gatacgccta
     6601 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     6661 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     6721 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     6781 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     6841 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     6901 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     6961 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt
     7021 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     7081 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     7141 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     7201 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     7261 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca
     7321 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     7381 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     7441 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     7501 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     7561 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     7621 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     7681 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     7741 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     7801 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     7861 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     7921 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     7981 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     8041 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     8101 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     8161 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     8221 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     8281 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     8341 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     8401 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     8461 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     8521 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     8581 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     8641 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     8701 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     8761 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gctctagcta
     8821 gaggtcgacc aattctcatg tttgacagct tatcatcgca gatccgggca acgttgttgc
     8881 cattgctgca ggcgcagaac tggtaggtat ggaagatcta tacattgaat caatattggc
     8941 aattagccat attagtcatt ggttatatag cataaatcaa tattggctat tggccattgc
     9001 atacgttgta tctatatcat aatatgtaca tttatattgg ctcatgtcca atatgaccgc
//
