LOCUS       sequence-274668-        6687 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 6687)
  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 6687)
  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: D92_Ld_360170
FEATURES             Location/Qualifiers
     source          1..6687
                     /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             1465..1482
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1534..1589
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1588..1607)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3825..3843)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3911..4015
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4016..4084,4085..4876)
                     /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(4234..4253)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5047..5635
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5536..5555
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5789..5806
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5923..5944
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5959..5964,5965..5982,5983..5989)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5997..6013
                     /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     6002..6024
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6021..6037
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6021..6037
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        6308..6687
                     /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 gcctctggag atcttcaaca gggcacatgg
      841 gacgtaagta ctctgctccc tggtagaggg gcagttcccc ctaattatga actgaccgta
      901 cagccaggca aatggacagt tagagaaaga gataacggta ggctgggcag acttttcgat
      961 tatgcagccg cccgggtggg tttggtttat gcaggggtgc ctagtgatga acatgcactc
     1021 tgccgaatgc catctagcag cactttgaag ttggccgccg ttaggactga tgacggggtg
     1081 cgcctggaat gccagactgt gtggttctct ccatccatgc tgagaagttt cgagaatgac
     1141 ggactcctgc tgagaaggtt tgatcctgca gagccacttt acagtgctga taatggcaga
     1201 tgtctggcta ctccatggcg tgtgcgtgag ttgtatgtgg cccgaatggg cgagggctcc
     1261 gtactcctga caggcattct ggtgggtgga gcaagtgaca ctcaggagcc atgctctgta
     1321 catcttgagg taaggcaccg ccctatcgcc agcgggaagg attcaagtgg cgcgcccggc
     1381 agcctgcacc acatcctgga cgcccagaag atgctgtgga accacaggga caggaacctg
     1441 cctcctctgg cccctctggg cccccaccac catcaccacc attgagtcta gacacgtgat
     1501 taattaagga tcccccgacc tcgacctctg gctaataaag gaaatttatt ttcattgcaa
     1561 tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt
     1621 tggtcgagat ccctcggaga tctctagcta gaggatcgat ccccgccccg gacgaactaa
     1681 acctgactac gacatctctg ccccttcttc gcggggcagt gcatgtaatc ccttcagttg
     1741 gttggtacaa cttgccaact gggccctgtt ccacatgtga cacggggggg gaccaaacac
     1801 aaaggggttc tctgactgta gttgacatcc ttataaatgg atgtgcacat ttgccaacac
     1861 tgagtggctt tcatcctgga gcagactttg cagtctgtgg actgcaacac aacattgcct
     1921 ttatgtgtaa ctcttggctg aagctcttac accaatgctg ggggacatgt acctcccagg
     1981 ggcccaggaa gactacggga ggctacacca acgtcaatca gaggggcctg tgtagctacc
     2041 gataagcgga ccctcaagag ggcattagca atagtgttta taaggccccc ttgttaaccc
     2101 taaacgggta gcatatgctt cccgggtagt agtatatact atccagacta accctaattc
     2161 aatagcatat gttacccaac gggaagcata tgctatcgaa ttagggttag taaaagggtc
     2221 ctaaggaaca gcgatatctc ccaccccatg agctgtcacg gttttattta catggggtca
     2281 ggattccacg agggtagtga accattttag tcacaagggc agtggctgaa gatcaaggag
     2341 cgggcagtga actctcctga atcttcgcct gcttcttcat tctccttcgt ttagctaata
     2401 gaataactgc tgagttgtga acagtaaggt gtatgtgagg tgctcgaaaa caaggtttca
     2461 ggtgacgccc ccagaataaa atttggacgg ggggttcagt ggtggcattg tgctatgaca
     2521 ccaatataac cctcacaaac cccttgggca ataaatacta gtgtaggaat gaaacattct
     2581 gaatatcttt aacaatagaa atccatgggg tggggacaag ccgtaaagac tggatgtcca
     2641 tctcacacga atttatggct atgggcaaca cataatccta gtgcaatatg atactggggt
     2701 tattaagatg tgtcccaggc agggaccaag acaggtgaac catgttgtta cactctattt
     2761 gtaacaaggg gaaagagagt ggacgccgac agcagcggac tccactggtt gtctctaaca
     2821 cccccgaaaa ttaaacgggg ctccacgcca atggggccca taaacaaaga caagtggcca
     2881 ctcttttttt tgaaattgtg gagtgggggc acgcgtcagc ccccacacgc cgccctgcgg
     2941 ttttggactg taaaataagg gtgtaataac ttggctgatt gtaaccccgc taaccactgc
     3001 ggtcaaacca cttgcccaca aaaccactaa tggcaccccg gggaatacct gcataagtag
     3061 gtgggcgggc caagataggg gcgcgattgc tgcgatctgg aggacaaatt acacacactt
     3121 gcgcctgagc gccaagcaca gggttgttgg tcctcatatt cacgaggtcg ctgagagcac
     3181 ggtgggctaa tgttgccatg ggtagcatat actacccaaa tatctggata gcatatgcta
     3241 tcctaatcta tatctgggta gcataggcta tcctaatcta tatctgggta gcatatgcta
     3301 tcctaatcta tatctgggta gtatatgcta tcctaattta tatctgggta gcataggcta
     3361 tcctaatcta tatctgggta gcatatgcta tcctaatcta tatctgggta gtatatgcta
     3421 tcctaatctg tatccgggta gcatatgcta tcctaataga gattagggta gtatatgcta
     3481 tcctaattta tatctgggta gcatatacta cccaaatatc tggatagcat atgctatcct
     3541 aatctatatc tgggtagcat atgctatcct aatctatatc tgggtagcat aggctatcct
     3601 aatctatatc tgggtagcat atgctatcct aatctatatc tgggtagtat atgctatcct
     3661 aatttatatc tgggtagcat aggctatcct aatctatatc tgggtagcat atgctatcct
     3721 aatctatatc tgggtagtat atgctatcct aatctgtatc cgggtagcat atgctatcct
     3781 catgataagc tgtcaaacat gagaattttc ttgaagacga aagggcctcg tgatacgcct
     3841 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     3901 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3961 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     4021 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4081 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     4141 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     4201 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt
     4261 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     4321 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     4381 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     4441 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     4501 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc
     4561 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     4621 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     4681 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     4741 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     4801 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     4861 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     4921 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     4981 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5041 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5101 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5161 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     5221 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5281 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5341 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5401 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5461 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5521 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5581 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5641 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     5701 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     5761 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     5821 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     5881 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     5941 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     6001 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agctctagct
     6061 agaggtcgac caattctcat gtttgacagc ttatcatcgc agatccgggc aacgttgttg
     6121 ccattgctgc aggcgcagaa ctggtaggta tggaagatct atacattgaa tcaatattgg
     6181 caattagcca tattagtcat tggttatata gcataaatca atattggcta ttggccattg
     6241 catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc aatatgaccg
     6301 ccatgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
     6361 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     6421 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     6481 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     6541 gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga cggtaaatgg
     6601 cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg gcagtacatc
     6661 tacgtattag tcatcgctat taccatg
//
