LOCUS       sequence-274859-        7047 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 7047)
  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 7047)
  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: D71_Ld_230810
FEATURES             Location/Qualifiers
     source          1..7047
                     /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             1825..1842
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1894..1949
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1948..1967)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4185..4203)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4271..4375
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4376..4444,4445..5236)
                     /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(4594..4613)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5407..5995
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5896..5915
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6149..6166
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6283..6304
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6319..6324,6325..6342,6343..6349)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6357..6373
                     /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     6362..6384
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6381..6397
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6381..6397
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        6668..7047
                     /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 gcccaagagg tcgccgccac tgccacagat
      841 gccgccggaa agcctgtggc tcagcacacg tccgttacaa ctgcaacctc tgaggaagag
      901 gcgagggcca gcaacgcagc agtgcaggcg gcagaggctg aagctgccgc aatgcgccaa
      961 ctggaagctg aggagcaagc aagacgcgaa gaagaggaag agttggccga gcgtcaacgg
     1021 cgacagcaag aagaagagag gagaagacag gaagaagagg aattggccga gagacagcgc
     1081 cgccaacaag aagaggaaag aagaagacag gaagaagagg agctcgcaga gcgccaacgt
     1141 cggcagcaag aggaagaaag ggcgaagcgg cgcattgata gcgtggatat gatccgctat
     1201 agcgccgagg aactgcccca aggatgtgct aaaagtatag aggagtacct gtttcgcgtc
     1261 cacgaacacc gtcaacacac agccgctgca gtaagggatg ccaggtccga attgaatgag
     1321 gctagggtgc atgcaccaaa gaaagtggcg gaactggaga aacggctcgc tgctcggcag
     1381 gatgacctgc tggtatacga tatccaaact ctgcgaatgt acggcgggtc agttcccaag
     1441 acatgtattg ccgaaagtct ggcttggctc gacgcacaac ctgacgtcag ctccatgagt
     1501 gaggcaaagc aacggttggt atatttcttc cgactcactt ctacccattg tagagctctg
     1561 tacctcactg cacgaggagt gatgcaatat atgatagaca gctatacccc cgtagtgaag
     1621 tcttacacta aatccgtgca gtattattgt gataaagcca gtgcgattta cgctgaattg
     1681 ctgcctgcat caggcaagcg gcctgacata cccatgatgg atttggccgg cgcgcccggc
     1741 agcctgcacc acatcctgga cgcccagaag atgctgtgga accacaggga caggaacctg
     1801 cctcctctgg cccctctggg cccccaccac catcaccacc attgagtcta gacacgtgat
     1861 taattaagga tcccccgacc tcgacctctg gctaataaag gaaatttatt ttcattgcaa
     1921 tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt
     1981 tggtcgagat ccctcggaga tctctagcta gaggatcgat ccccgccccg gacgaactaa
     2041 acctgactac gacatctctg ccccttcttc gcggggcagt gcatgtaatc ccttcagttg
     2101 gttggtacaa cttgccaact gggccctgtt ccacatgtga cacggggggg gaccaaacac
     2161 aaaggggttc tctgactgta gttgacatcc ttataaatgg atgtgcacat ttgccaacac
     2221 tgagtggctt tcatcctgga gcagactttg cagtctgtgg actgcaacac aacattgcct
     2281 ttatgtgtaa ctcttggctg aagctcttac accaatgctg ggggacatgt acctcccagg
     2341 ggcccaggaa gactacggga ggctacacca acgtcaatca gaggggcctg tgtagctacc
     2401 gataagcgga ccctcaagag ggcattagca atagtgttta taaggccccc ttgttaaccc
     2461 taaacgggta gcatatgctt cccgggtagt agtatatact atccagacta accctaattc
     2521 aatagcatat gttacccaac gggaagcata tgctatcgaa ttagggttag taaaagggtc
     2581 ctaaggaaca gcgatatctc ccaccccatg agctgtcacg gttttattta catggggtca
     2641 ggattccacg agggtagtga accattttag tcacaagggc agtggctgaa gatcaaggag
     2701 cgggcagtga actctcctga atcttcgcct gcttcttcat tctccttcgt ttagctaata
     2761 gaataactgc tgagttgtga acagtaaggt gtatgtgagg tgctcgaaaa caaggtttca
     2821 ggtgacgccc ccagaataaa atttggacgg ggggttcagt ggtggcattg tgctatgaca
     2881 ccaatataac cctcacaaac cccttgggca ataaatacta gtgtaggaat gaaacattct
     2941 gaatatcttt aacaatagaa atccatgggg tggggacaag ccgtaaagac tggatgtcca
     3001 tctcacacga atttatggct atgggcaaca cataatccta gtgcaatatg atactggggt
     3061 tattaagatg tgtcccaggc agggaccaag acaggtgaac catgttgtta cactctattt
     3121 gtaacaaggg gaaagagagt ggacgccgac agcagcggac tccactggtt gtctctaaca
     3181 cccccgaaaa ttaaacgggg ctccacgcca atggggccca taaacaaaga caagtggcca
     3241 ctcttttttt tgaaattgtg gagtgggggc acgcgtcagc ccccacacgc cgccctgcgg
     3301 ttttggactg taaaataagg gtgtaataac ttggctgatt gtaaccccgc taaccactgc
     3361 ggtcaaacca cttgcccaca aaaccactaa tggcaccccg gggaatacct gcataagtag
     3421 gtgggcgggc caagataggg gcgcgattgc tgcgatctgg aggacaaatt acacacactt
     3481 gcgcctgagc gccaagcaca gggttgttgg tcctcatatt cacgaggtcg ctgagagcac
     3541 ggtgggctaa tgttgccatg ggtagcatat actacccaaa tatctggata gcatatgcta
     3601 tcctaatcta tatctgggta gcataggcta tcctaatcta tatctgggta gcatatgcta
     3661 tcctaatcta tatctgggta gtatatgcta tcctaattta tatctgggta gcataggcta
     3721 tcctaatcta tatctgggta gcatatgcta tcctaatcta tatctgggta gtatatgcta
     3781 tcctaatctg tatccgggta gcatatgcta tcctaataga gattagggta gtatatgcta
     3841 tcctaattta tatctgggta gcatatacta cccaaatatc tggatagcat atgctatcct
     3901 aatctatatc tgggtagcat atgctatcct aatctatatc tgggtagcat aggctatcct
     3961 aatctatatc tgggtagcat atgctatcct aatctatatc tgggtagtat atgctatcct
     4021 aatttatatc tgggtagcat aggctatcct aatctatatc tgggtagcat atgctatcct
     4081 aatctatatc tgggtagtat atgctatcct aatctgtatc cgggtagcat atgctatcct
     4141 catgataagc tgtcaaacat gagaattttc ttgaagacga aagggcctcg tgatacgcct
     4201 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     4261 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     4321 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     4381 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4441 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     4501 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     4561 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt
     4621 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     4681 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     4741 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     4801 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     4861 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc
     4921 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     4981 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     5041 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     5101 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     5161 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     5221 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     5281 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     5341 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5401 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5461 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5521 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     5581 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5641 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5701 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5761 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5821 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5881 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5941 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6001 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     6061 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     6121 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     6181 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     6241 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     6301 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     6361 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agctctagct
     6421 agaggtcgac caattctcat gtttgacagc ttatcatcgc agatccgggc aacgttgttg
     6481 ccattgctgc aggcgcagaa ctggtaggta tggaagatct atacattgaa tcaatattgg
     6541 caattagcca tattagtcat tggttatata gcataaatca atattggcta ttggccattg
     6601 catacgttgt atctatatca tgatatgtac atttatattg gctcatgtcc aatatgaccg
     6661 ccatgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
     6721 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     6781 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     6841 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     6901 gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga cggtaaatgg
     6961 cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg gcagtacatc
     7021 tacgtattag tcatcgctat taccatg
//
