LOCUS       sequence-262382-        7935 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 7935)
  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 7935)
  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: D90_Ld_350740
FEATURES             Location/Qualifiers
     source          1..7935
                     /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             3099..3116
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3168..3223
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3222..3241)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5459..5477)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5545..5649
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5650..5718,5719..6510)
                     /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(5868..5887)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6681..7269
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7170..7189
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7423..7440
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7557..7578
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7593..7598,7599..7616,7617..7623)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7631..7647
                     /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     7636..7658
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7655..7671
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7655..7671
                     /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 gcacgccagc ccccgctcct ggagaatccc cgtctaagac cgctctcaat gcagaagtct
     1261 ctgtctctac cgctcccccg ccaactacat ctagtagtgc ggtgccaagc ggcagcgcta
     1321 ccgaccaaca cagaggggcc gatgcacctt atagaagcct gtatccacaa gttaagctcc
     1381 gggccacaac ccaggaaagg ccatataaac gaagaggctt tgttgccttc gaagaagcag
     1441 agacaggggt agagcaacgg cctagactcg aagatggctc cgcccaagtt attgctcacc
     1501 tgcccaccgc agcagaaact ggcggggccg caattctgcc tggtcataga agaagtcacg
     1561 ctcaaacttc acttgctact caagagggcg gctgcgggca gttgagcgat gcagctgaag
     1621 gcaatgaggc ccaacaattt ctggacgaag tacaaaggca ggtcgacgta gctagacatg
     1681 ctaggaacca ggccatccct ttcccccaga aacctgaagc gaattcccca gcttttcgaa
     1741 ggataaagag acacgccaaa atgaagattg aggtgcccga ccccgactat cccaccttcg
     1801 ctcgcaagga caaagtcatg caactgcccc caccccacga acatccatgg gttcggaaga
     1861 acacacctat aggacccttt attgtacacg gggatggtca gattcatcag acaggcggca
     1921 ctggccaggt ggatttcgat gattcctctg tgaatgaaaa gctgcctcag tcctaccgcg
     1981 ggcttcagca ccggtccacc cttcaacggc aattgcctca ggacaatgga agggtgcttc
     2041 aagaggccgt tataaaacac agcttcacac ttacgggcag aggcctgttt gctactcgac
     2101 ggatagcaaa aggggaaacc atcatgattg ttcagagtac ggcacgcaac gtaggagtaa
     2161 agggagagct tcaacgtttg gaagagatgt gcacagacat actgatcgct tgtagagatg
     2221 gggacgcccg tgcttgcgac tacttgcatg attggatcct gactggacag ccaagttccc
     2281 tgctcgagca ctggccagcc gcatccactt cccgagtcat ggacgccatc ggcggtgcgg
     2341 atgtgcttag agcgcttgag ctgcatccga tccacattgc gcgcatggca gccattattg
     2401 atctgaatag tttcctggtt gagtccagct tttccgaaag gaaaggcatg gcatactttc
     2461 ccgaagcagg gttcctgaac cactcctgcg ctccgaatgc cacttatgac attatgcctg
     2521 aacatgtttt ccgggagaca gattattacc tggacgaagc aaccagggcg acggcagcgg
     2581 aatccggaga agaagaggac gtgcagggta ctgtgatggc cgcagatgat gcaggtacat
     2641 cttcaacagc tgcgccatgt ccggcacgtg ccgctcagcc cccagccgca gtgggatata
     2701 ttagctcctc acaggttctg gtctgtgacg gcgtacccga aggacaaaga cacgttgcgt
     2761 tcttcggtca tgaagaccga ctggccgcct accccgggct gacggaagcc gatgctccgg
     2821 tctacttgtt ttgttgtaga gccgagaagg aaatagaggc tggtgaggaa attgtgatta
     2881 gctacgtgcc cccgcagtgg tcatttgata atcgacagta cgtgctgcat gatcgctaca
     2941 ggttttggtg taagtgtccg aaatgcagcc ccgtgcttga taagaagtac gccagggtgc
     3001 ctggcgcgcc cggcagcctg caccacatcc tggacgccca gaagatgctg tggaaccaca
     3061 gggacaggaa cctgcctcct ctggcccctc tgggccccca ccaccatcac caccattgag
     3121 tctagacacg tgattaatta aggatccccc gacctcgacc tctggctaat aaaggaaatt
     3181 tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg
     3241 gagggcaaat catttggtcg agatccctcg gagatctcta gctagaggat cgatccccgc
     3301 cccggacgaa ctaaacctga ctacgacatc tctgcccctt cttcgcgggg cagtgcatgt
     3361 aatcccttca gttggttggt acaacttgcc aactgggccc tgttccacat gtgacacggg
     3421 gggggaccaa acacaaaggg gttctctgac tgtagttgac atccttataa atggatgtgc
     3481 acatttgcca acactgagtg gctttcatcc tggagcagac tttgcagtct gtggactgca
     3541 acacaacatt gcctttatgt gtaactcttg gctgaagctc ttacaccaat gctgggggac
     3601 atgtacctcc caggggccca ggaagactac gggaggctac accaacgtca atcagagggg
     3661 cctgtgtagc taccgataag cggaccctca agagggcatt agcaatagtg tttataaggc
     3721 ccccttgtta accctaaacg ggtagcatat gcttcccggg tagtagtata tactatccag
     3781 actaacccta attcaatagc atatgttacc caacgggaag catatgctat cgaattaggg
     3841 ttagtaaaag ggtcctaagg aacagcgata tctcccaccc catgagctgt cacggtttta
     3901 tttacatggg gtcaggattc cacgagggta gtgaaccatt ttagtcacaa gggcagtggc
     3961 tgaagatcaa ggagcgggca gtgaactctc ctgaatcttc gcctgcttct tcattctcct
     4021 tcgtttagct aatagaataa ctgctgagtt gtgaacagta aggtgtatgt gaggtgctcg
     4081 aaaacaaggt ttcaggtgac gcccccagaa taaaatttgg acggggggtt cagtggtggc
     4141 attgtgctat gacaccaata taaccctcac aaaccccttg ggcaataaat actagtgtag
     4201 gaatgaaaca ttctgaatat ctttaacaat agaaatccat ggggtgggga caagccgtaa
     4261 agactggatg tccatctcac acgaatttat ggctatgggc aacacataat cctagtgcaa
     4321 tatgatactg gggttattaa gatgtgtccc aggcagggac caagacaggt gaaccatgtt
     4381 gttacactct atttgtaaca aggggaaaga gagtggacgc cgacagcagc ggactccact
     4441 ggttgtctct aacacccccg aaaattaaac ggggctccac gccaatgggg cccataaaca
     4501 aagacaagtg gccactcttt tttttgaaat tgtggagtgg gggcacgcgt cagcccccac
     4561 acgccgccct gcggttttgg actgtaaaat aagggtgtaa taacttggct gattgtaacc
     4621 ccgctaacca ctgcggtcaa accacttgcc cacaaaacca ctaatggcac cccggggaat
     4681 acctgcataa gtaggtgggc gggccaagat aggggcgcga ttgctgcgat ctggaggaca
     4741 aattacacac acttgcgcct gagcgccaag cacagggttg ttggtcctca tattcacgag
     4801 gtcgctgaga gcacggtggg ctaatgttgc catgggtagc atatactacc caaatatctg
     4861 gatagcatat gctatcctaa tctatatctg ggtagcatag gctatcctaa tctatatctg
     4921 ggtagcatat gctatcctaa tctatatctg ggtagtatat gctatcctaa tttatatctg
     4981 ggtagcatag gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg
     5041 ggtagtatat gctatcctaa tctgtatccg ggtagcatat gctatcctaa tagagattag
     5101 ggtagtatat gctatcctaa tttatatctg ggtagcatat actacccaaa tatctggata
     5161 gcatatgcta tcctaatcta tatctgggta gcatatgcta tcctaatcta tatctgggta
     5221 gcataggcta tcctaatcta tatctgggta gcatatgcta tcctaatcta tatctgggta
     5281 gtatatgcta tcctaattta tatctgggta gcataggcta tcctaatcta tatctgggta
     5341 gcatatgcta tcctaatcta tatctgggta gtatatgcta tcctaatctg tatccgggta
     5401 gcatatgcta tcctcatgat aagctgtcaa acatgagaat tttcttgaag acgaaagggc
     5461 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     5521 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     5581 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     5641 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     5701 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     5761 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     5821 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     5881 gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     5941 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     6001 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     6061 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     6121 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     6181 accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     6241 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     6301 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     6361 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     6421 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     6481 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     6541 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     6601 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     6661 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     6721 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     6781 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     6841 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     6901 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     6961 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     7021 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     7081 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     7141 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     7201 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     7261 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     7321 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     7381 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     7441 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     7501 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     7561 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     7621 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     7681 gccaagctct agctagaggt cgaccaattc tcatgtttga cagcttatca tcgcagatcc
     7741 gggcaacgtt gttgccattg ctgcaggcgc agaactggta ggtatggaag atctatacat
     7801 tgaatcaata ttggcaatta gccatattag tcattggtta tatagcataa atcaatattg
     7861 gctattggcc attgcatacg ttgtatctat atcataatat gtacatttat attggctcat
     7921 gtccaatatg accgc
//
