LOCUS       sequence-274882-        7602 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 7602)
  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 7602)
  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: D61_Ld_351340
FEATURES             Location/Qualifiers
     source          1..7602
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     201..225
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     706..725
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      748..757
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2380..2397
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2449..2504
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2503..2522)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4740..4758)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4826..4930
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4931..4999,5000..5791)
                     /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(5149..5168)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5962..6550
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6451..6470
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6704..6721
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6838..6859
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6874..6879,6880..6897,6898..6904)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6912..6928
                     /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     6917..6939
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6936..6952
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6936..6952
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7223..7602
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta caccaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcctcac tctcttccgc
      241 atcgctgtct gcgagggcca gctgttgggc tcgcggttga ggacaaactc ttcgcggtct
      301 ttccagtact cttggatcgg aaacccgtcg gcctccgaac ggtactccgc caccgaggga
      361 cctgagcgag tccgcatcga ccggatcgga aaacctctcg agaaaggcgt ctaaccagtc
      421 acagtcgcaa ggtaggctga gcaccgtggc gggcggcagc gggtggcggt cggggttgtt
      481 tctggcggag gtgctgctga tgatgtaatt aaagtaggcg gtcttgagac ggcggatggt
      541 cgaggtgagg tgtggcaggc ttgagatcca gctgttgggg tgagtactcc ctctcaaaag
      601 cgggcattac ttctgcgcta agattgtcag tttccaaaaa cgaggaggat ttgatattca
      661 cctggcccga tctggccata cacttgagtg acaatgacat ccactttgcc tttctctcca
      721 caggtgtcca ctcccaggtc caagtttgcc accatggagt ttcagaccca ggtactcatg
      781 tccctgctgc tctgcatgtc tggtgcggcc gccgtggaag atgtgggaga ggatgccagc
      841 gccgtgctgg aactgatcag cgagggcagc accaccacct acaagatccc cgacagcatg
      901 gtggtgctga acaacgccaa cttcgagagc tacctgttcc ccagcaagcg ggccaccccc
      961 agagcctttc tggtgctgtg ctacagcccc tggtgccccc actgcaagag cctgctgccc
     1021 cagttcctga acgccagcat gcagctggac ctgatgaagg tgccccacag caacttcgcc
     1081 gtggtggacg tgcagaaaaa caccgccgtg tccgagtact tcgacgtgga acggttcccc
     1141 accctgctgt acaccaccgg caagggccgg cagtggcacc tgtacgaggg cggcaatacc
     1201 cagcagggct tcatgcagtt cagcacctac ctgcagaacg ccatggacac cggcagcttc
     1261 agcgaggacg tgaccgacgt gtcccacttc aacgaggtgg aagagaagtc cggcaccacc
     1321 cgggtgccct gctacgtgta cgtgcccgcc acaagcagca gcgcccctga gagccagaga
     1381 accgcccact ggtcccacgc cattgacggc gctgccagcg tgtccaacat cagattcgcc
     1441 gtgatctacg agaagtccca ggccgaggga tgggccgagc acgccagcga caagtacaag
     1501 aaagtggtgg aaaaggccaa ggcctgcgtg gccgctggca aggcttctgg acctggcgga
     1561 gaggccctgg tggtgttcag cgaccggtac agagagcccc actgctacag cggcccctgg
     1621 gtggaagaaa gaagcgtggc cagaagcagc aagcaccgga ccagacaggt ggacgccgac
     1681 accctgacca tgtctaccag cctggaaaac ttcctggccc tgcacggctt ccacgccgtc
     1741 gaggatgcct ccagcgccat gttcgccacc ctggcctact accccaagaa ctacctgggc
     1801 gtcgtgatga ccaaccggcc catcgacgac aaggacatgg acttcgtgcc cgtgctgaga
     1861 gagatcacac aggccgagaa cgccgccctg gaaaagaagc acggcagcga cctgcccatc
     1921 gaggaagaga tgcggacccc cagagtgtcc tggtcctaca tcgacgtggt ggaatacgaa
     1981 gtgtggcgga gcagatacga catcgagctg gaccagctgc ccgccgtgat gatcatcgac
     2041 accaagcggg accggttctt caagatgaga acccacgtgc ccagattcga ggccatcaag
     2101 atggacaccc cctggaaagt gggcggcgag cagcagcagc tgatcgccca gttcgcccag
     2161 gacgtgctgg ccgacgccta caaggcccag aaactgtccg tggctggcgc cgtggccgaa
     2221 tacctgagcc actaccccgg cttcgccctg atgtacgagg ccctgaacta cgaggacttc
     2281 gtgggcgcgc ccggcagcct gcaccacatc ctggacgccc agaagatgct gtggaaccac
     2341 agggacagga acctgcctcc tctggcccct ctgggccccc accaccatca ccaccattga
     2401 gtctagacac gtgattaatt aaggatcccc cgacctcgac ctctggctaa taaaggaaat
     2461 ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg
     2521 ggagggcaaa tcatttggtc gagatccctc ggagatctct agctagagga tcgatccccg
     2581 ccccggacga actaaacctg actacgacat ctctgcccct tcttcgcggg gcagtgcatg
     2641 taatcccttc agttggttgg tacaacttgc caactgggcc ctgttccaca tgtgacacgg
     2701 ggggggacca aacacaaagg ggttctctga ctgtagttga catccttata aatggatgtg
     2761 cacatttgcc aacactgagt ggctttcatc ctggagcaga ctttgcagtc tgtggactgc
     2821 aacacaacat tgcctttatg tgtaactctt ggctgaagct cttacaccaa tgctggggga
     2881 catgtacctc ccaggggccc aggaagacta cgggaggcta caccaacgtc aatcagaggg
     2941 gcctgtgtag ctaccgataa gcggaccctc aagagggcat tagcaatagt gtttataagg
     3001 cccccttgtt aaccctaaac gggtagcata tgcttcccgg gtagtagtat atactatcca
     3061 gactaaccct aattcaatag catatgttac ccaacgggaa gcatatgcta tcgaattagg
     3121 gttagtaaaa gggtcctaag gaacagcgat atctcccacc ccatgagctg tcacggtttt
     3181 atttacatgg ggtcaggatt ccacgagggt agtgaaccat tttagtcaca agggcagtgg
     3241 ctgaagatca aggagcgggc agtgaactct cctgaatctt cgcctgcttc ttcattctcc
     3301 ttcgtttagc taatagaata actgctgagt tgtgaacagt aaggtgtatg tgaggtgctc
     3361 gaaaacaagg tttcaggtga cgcccccaga ataaaatttg gacggggggt tcagtggtgg
     3421 cattgtgcta tgacaccaat ataaccctca caaacccctt gggcaataaa tactagtgta
     3481 ggaatgaaac attctgaata tctttaacaa tagaaatcca tggggtgggg acaagccgta
     3541 aagactggat gtccatctca cacgaattta tggctatggg caacacataa tcctagtgca
     3601 atatgatact ggggttatta agatgtgtcc caggcaggga ccaagacagg tgaaccatgt
     3661 tgttacactc tatttgtaac aaggggaaag agagtggacg ccgacagcag cggactccac
     3721 tggttgtctc taacaccccc gaaaattaaa cggggctcca cgccaatggg gcccataaac
     3781 aaagacaagt ggccactctt ttttttgaaa ttgtggagtg ggggcacgcg tcagccccca
     3841 cacgccgccc tgcggttttg gactgtaaaa taagggtgta ataacttggc tgattgtaac
     3901 cccgctaacc actgcggtca aaccacttgc ccacaaaacc actaatggca ccccggggaa
     3961 tacctgcata agtaggtggg cgggccaaga taggggcgcg attgctgcga tctggaggac
     4021 aaattacaca cacttgcgcc tgagcgccaa gcacagggtt gttggtcctc atattcacga
     4081 ggtcgctgag agcacggtgg gctaatgttg ccatgggtag catatactac ccaaatatct
     4141 ggatagcata tgctatccta atctatatct gggtagcata ggctatccta atctatatct
     4201 gggtagcata tgctatccta atctatatct gggtagtata tgctatccta atttatatct
     4261 gggtagcata ggctatccta atctatatct gggtagcata tgctatccta atctatatct
     4321 gggtagtata tgctatccta atctgtatcc gggtagcata tgctatccta atagagatta
     4381 gggtagtata tgctatccta atttatatct gggtagcata tactacccaa atatctggat
     4441 agcatatgct atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt
     4501 agcataggct atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt
     4561 agtatatgct atcctaattt atatctgggt agcataggct atcctaatct atatctgggt
     4621 agcatatgct atcctaatct atatctgggt agtatatgct atcctaatct gtatccgggt
     4681 agcatatgct atcctcatga taagctgtca aacatgagaa ttttcttgaa gacgaaaggg
     4741 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     4801 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     4861 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     4921 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     4981 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     5041 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     5101 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     5161 ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     5221 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     5281 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     5341 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     5401 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     5461 caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     5521 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     5581 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     5641 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     5701 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     5761 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     5821 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     5881 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     5941 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     6001 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     6061 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     6121 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     6181 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     6241 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     6301 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6361 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6421 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     6481 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     6541 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     6601 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     6661 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     6721 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     6781 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     6841 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     6901 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     6961 cgccaagctc tagctagagg tcgaccaatt ctcatgtttg acagcttatc atcgcagatc
     7021 cgggcaacgt tgttgccatt gctgcaggcg cagaactggt aggtatggaa gatctataca
     7081 ttgaatcaat attggcaatt agccatatta gtcattggtt atatagcata aatcaatatt
     7141 ggctattggc cattgcatac gttgtatcta tatcataata tgtacattta tattggctca
     7201 tgtccaatat gaccgccatg ttgacattga ttattgacta gttattaata gtaatcaatt
     7261 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     7321 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     7381 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     7441 actgcccact tggcagtaca tcaagtgtat catatgccaa gtccgccccc tattgacgtc
     7501 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttacg ggactttcct
     7561 acttggcagt acatctacgt attagtcatc gctattacca tg
//
