LOCUS       sequence-274679-        7017 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 7017)
  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 7017)
  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: D87_Ld_301610
FEATURES             Location/Qualifiers
     source          1..7017
                     /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             1795..1812
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1864..1919
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1918..1937)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4155..4173)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4241..4345
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4346..4414,4415..5206)
                     /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(4564..4583)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5377..5965
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5866..5885
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6119..6136
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6253..6274
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6289..6294,6295..6312,6313..6319)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6327..6343
                     /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     6332..6354
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6351..6367
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6351..6367
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        6638..7017
                     /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 gccgctccac ctgcctcagc cgaagccgcc
      841 gttcccaact cagctgccag aactgtgccc cctttcgagc agttctttgc gcagtccttc
      901 caagtgagtg tgatgactca gtcctctggc gtgtttaatg caaccctgag aatgagggcg
      961 tcacccaact ttcccgaacg cgttcagggc gagctgattc ctattggtga agccaaactt
     1021 aacagccaga gacagacgtt gctcccccac ttccaaagaa cccgttccct ggaagatgac
     1081 cgtctttccg cactgtcctg ctctatggca gctgggttcg gaagaacaaa taggaagaag
     1141 aaaggtagca ctgatgagag tgggtctggt gaatttgcag ttgcgggaaa ggaagaacgc
     1201 ccaagcctgc tgatggttga tctccacttg agacactccg atgccatgca ggggacagcc
     1261 tctgtgtact acctgccggc agatctgatg gccttgcgtg ctcagaggga aggcatcaag
     1321 gaagggagcg caccaccatc agctacagtg gaattcgagt tcagaagtga agtggatcat
     1381 atgtccggga acccactgag cgatgttcag gcactcgcac acgtatccag tgccgtggtc
     1441 acaatgggtg gacagtcagg cgcctctaag agcagcagtg ttgccgggga gaaggtggat
     1501 gacgggagcg ctgccgccac aaggcagggt gggatcattt tcaggtggat aactgaacac
     1561 gaattttccg cctacgtaac tattccgatt actgacaaaa caggggctac atcaactcat
     1621 atggagcaaa tatgggtgta tgggtatgct gtcccatcca agagtgtgtt tgataggaag
     1681 cagagagaat tgccgctcgg cgcgcccggc agcctgcacc acatcctgga cgcccagaag
     1741 atgctgtgga accacaggga caggaacctg cctcctctgg cccctctggg cccccaccac
     1801 catcaccacc attgagtcta gacacgtgat taattaagga tcccccgacc tcgacctctg
     1861 gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac
     1921 tcggaaggac atatgggagg gcaaatcatt tggtcgagat ccctcggaga tctctagcta
     1981 gaggatcgat ccccgccccg gacgaactaa acctgactac gacatctctg ccccttcttc
     2041 gcggggcagt gcatgtaatc ccttcagttg gttggtacaa cttgccaact gggccctgtt
     2101 ccacatgtga cacggggggg gaccaaacac aaaggggttc tctgactgta gttgacatcc
     2161 ttataaatgg atgtgcacat ttgccaacac tgagtggctt tcatcctgga gcagactttg
     2221 cagtctgtgg actgcaacac aacattgcct ttatgtgtaa ctcttggctg aagctcttac
     2281 accaatgctg ggggacatgt acctcccagg ggcccaggaa gactacggga ggctacacca
     2341 acgtcaatca gaggggcctg tgtagctacc gataagcgga ccctcaagag ggcattagca
     2401 atagtgttta taaggccccc ttgttaaccc taaacgggta gcatatgctt cccgggtagt
     2461 agtatatact atccagacta accctaattc aatagcatat gttacccaac gggaagcata
     2521 tgctatcgaa ttagggttag taaaagggtc ctaaggaaca gcgatatctc ccaccccatg
     2581 agctgtcacg gttttattta catggggtca ggattccacg agggtagtga accattttag
     2641 tcacaagggc agtggctgaa gatcaaggag cgggcagtga actctcctga atcttcgcct
     2701 gcttcttcat tctccttcgt ttagctaata gaataactgc tgagttgtga acagtaaggt
     2761 gtatgtgagg tgctcgaaaa caaggtttca ggtgacgccc ccagaataaa atttggacgg
     2821 ggggttcagt ggtggcattg tgctatgaca ccaatataac cctcacaaac cccttgggca
     2881 ataaatacta gtgtaggaat gaaacattct gaatatcttt aacaatagaa atccatgggg
     2941 tggggacaag ccgtaaagac tggatgtcca tctcacacga atttatggct atgggcaaca
     3001 cataatccta gtgcaatatg atactggggt tattaagatg tgtcccaggc agggaccaag
     3061 acaggtgaac catgttgtta cactctattt gtaacaaggg gaaagagagt ggacgccgac
     3121 agcagcggac tccactggtt gtctctaaca cccccgaaaa ttaaacgggg ctccacgcca
     3181 atggggccca taaacaaaga caagtggcca ctcttttttt tgaaattgtg gagtgggggc
     3241 acgcgtcagc ccccacacgc cgccctgcgg ttttggactg taaaataagg gtgtaataac
     3301 ttggctgatt gtaaccccgc taaccactgc ggtcaaacca cttgcccaca aaaccactaa
     3361 tggcaccccg gggaatacct gcataagtag gtgggcgggc caagataggg gcgcgattgc
     3421 tgcgatctgg aggacaaatt acacacactt gcgcctgagc gccaagcaca gggttgttgg
     3481 tcctcatatt cacgaggtcg ctgagagcac ggtgggctaa tgttgccatg ggtagcatat
     3541 actacccaaa tatctggata gcatatgcta tcctaatcta tatctgggta gcataggcta
     3601 tcctaatcta tatctgggta gcatatgcta tcctaatcta tatctgggta gtatatgcta
     3661 tcctaattta tatctgggta gcataggcta tcctaatcta tatctgggta gcatatgcta
     3721 tcctaatcta tatctgggta gtatatgcta tcctaatctg tatccgggta gcatatgcta
     3781 tcctaataga gattagggta gtatatgcta tcctaattta tatctgggta gcatatacta
     3841 cccaaatatc tggatagcat atgctatcct aatctatatc tgggtagcat atgctatcct
     3901 aatctatatc tgggtagcat aggctatcct aatctatatc tgggtagcat atgctatcct
     3961 aatctatatc tgggtagtat atgctatcct aatttatatc tgggtagcat aggctatcct
     4021 aatctatatc tgggtagcat atgctatcct aatctatatc tgggtagtat atgctatcct
     4081 aatctgtatc cgggtagcat atgctatcct catgataagc tgtcaaacat gagaattttc
     4141 ttgaagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     4201 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     4261 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4321 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     4381 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     4441 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     4501 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     4561 tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg
     4621 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     4681 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     4741 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     4801 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     4861 aaacgacgag cgtgacacca cgatgcctgc agcaatggca acaacgttgc gcaaactatt
     4921 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     4981 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     5041 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     5101 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     5161 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     5221 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     5281 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     5341 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     5401 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     5461 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     5521 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     5581 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     5641 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     5701 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     5761 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     5821 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     5881 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     5941 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     6001 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     6061 ccgtattacc gcctttgagt gagctgatac cgctcgtcgc agccgaacga ccgagcgcag
     6121 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     6181 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     6241 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
     6301 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
     6361 ctatgaccat gattacgcca agctctagct agaggtcgac caattctcat gtttgacagc
     6421 ttatcatcgc agatccgggc aacgttgttg ccattgctgc aggcgcagaa ctggtaggta
     6481 tggaagatct atacattgaa tcaatattgg caattagcca tattagtcat tggttatata
     6541 gcataaatca atattggcta ttggccattg catacgttgt atctatatca taatatgtac
     6601 atttatattg gctcatgtcc aatatgaccg ccatgttgac attgattatt gactagttat
     6661 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
     6721 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
     6781 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
     6841 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtccg
     6901 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
     6961 ttacgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatg
//
