LOCUS       sequence-274879-        6496 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 6496)
  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 6496)
  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: D64_Ld_010010
FEATURES             Location/Qualifiers
     source          1..6496
                     /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             1570..1587
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1639..1694
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1693..1712)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3634..3652)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3720..3824
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3825..3893,3894..4685)
                     /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(4043..4062)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4856..5444
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5345..5364
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5598..5615
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5732..5753
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5768..5773,5774..5791,5792..5798)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5806..5822
                     /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     5811..5833
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5830..5846
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5830..5846
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        6117..6496
                     /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 gcctactcta ccacttacgc cgcgcgctct
      841 gcagggtctg gagctttcgc cttcgaccca ctcgagatcg acttttgtaa actgcaggac
      901 gcggcttcac ataccgccat gtccaccagc gcatcaatcc aggaagctct tcaaagcgct
      961 tatcagcgtg aaggaatatc cggaaacatc cctcttggcg gaatgacaga gcaggcagtg
     1021 tgcgccggta cacagtgtga atggcgctgg taccggggcc tgtttaggac cacccgccct
     1081 cttttcctta ggggagtgaa ttattggggc gttaacgacg atagcactcc cgtggaccgt
     1141 accacagcag tccccggcag ttatactaat ttcgagacaa actacccacg cagctgggga
     1201 aatctcagct actggggaaa gaggctcgtg gtgatgcagc cctcaggcaa atgggttaat
     1261 acagaagttt ctacatcctt tacccatttc ttgtgcgagt actacgttta cgcagacagc
     1321 gacggtgtga acaggctcgt gccgactttt cccggcggat ctgcaattcc gttcgtctcc
     1381 agcagtggac atacatacac ctactgtggt aagaaggtcg aggtggactc tcaatcccgc
     1441 tggatcttgc ctaaatgtaa tgaaaccttt cccggcgcgc ccggcagcct gcaccacatc
     1501 ctggacgccc agaagatgct gtggaaccac agggacagga acctgcctcc tctggcccct
     1561 ctgggccccc accaccatca ccaccattga gtctagacac gtgattaatt aaggatcccc
     1621 cgacctcgac ctctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt
     1681 ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttggtc gagatccctc
     1741 ggagatctct agctagagga tcgatccccg ccccggacga actaaacctg actacgacat
     1801 ctctgcccct tcttcgcggg gcagtgcatg taatcccttc agttggttgg tacaacttgc
     1861 caactgggcc ctgttccaca tgtgacacgg ggggggacca aacacaaagg ggttctctga
     1921 ctgtagttga catccttata aatggatgtg cacatttgcc aacactgagt ggctttcatc
     1981 ctggagcaga ctttgcagtc tgtggactgc aacacaacat tgcctttatg tgtaactctt
     2041 ggctgaagct cttacaccaa tgctggggga catgtacctc ccaggggccc aggaagacta
     2101 cgggaggcta caccaacgtc aatcagaggg gcctgtgtag ctaccgataa gcggaccctc
     2161 aagagggcat tagcaatagt gtttataagg cccccttgtt aaccctaaac gggtagcata
     2221 tgcttcccgg gtagtagtat atactatcca gactaaccct aattcaatag catatgttac
     2281 ccaacgggaa gcatatgcta tcgaattagg gttagtaaaa gggtcctaag gaacagcgat
     2341 atctcccacc ccatgagctg tcacggtttt atttacatgg ggtcaggatt ccacgagggt
     2401 agtgaaccat tttagtcaca agggcagtgg ctgaagatca aggagcgggc agtgaactct
     2461 cctgaatctt cgcctgcttc ttcattctcc ttcgtttagc taatagaata actgctgagt
     2521 tgtgaacagt aaggtgtatg tgaggtgctc gaaaacaagg tttcaggtga cgcccccaga
     2581 ataaaatttg gacggggggt tcagtggtgg cattgtgcta tgacaccaat ataaccctca
     2641 caaacccctt gggcaataaa tactagtgta ggaatgaaac attctgaata tctttaacaa
     2701 tagaaatcca tggggtgggg acaagccgta aagactggat gtccatctca cacgaattta
     2761 tggctatggg caacacataa tcctagtgca atatgatact ggggttatta agatgtgtcc
     2821 caggcaggga ccaagacagg tgaaccatgt tgttacactc tatttgtaac aaggggaaag
     2881 agagtggacg ccgacagcag cggactccac tggttgtctc taacaccccc gaaaattaaa
     2941 cggggctcca cgccaatggg gcccataaac aaagacaagt ggccactctt ttttttgaaa
     3001 ttgtggagtg ggggcacgcg tcagccccca cacgccgccc tgcggttttg gactgtaaaa
     3061 taagggtgta ataacttggc tgattgtaac cccgctaacc actgcggtca aaccacttgc
     3121 ccacaaaacc actaatggca ccccggggaa tacctgcata agtaggtggg cgggccaaga
     3181 taggggcgcg attgctgcga tctggaggac aaattacaca cacttgcgcc tgagcgccaa
     3241 gcacagggtt gttggtcctc atattcacga ggtcgctgag agcacggtgg gctaatgttg
     3301 ccatgggtag catatactac ccaaatatct ggatagcata tgctatccta atctatatct
     3361 gggtagcata ggctatccta atctatatct gggtagcata tgctatccta atctatatct
     3421 gggtagtata tgctatccta atttatatct gggtagyata kgctatccta atytatatct
     3481 gggtagcata ggctatccta atctatatct gggtagcata tgctatccta atctatatct
     3541 gggtagtata tgctatccta atctgtatcc gggtagcata tgctatcctc atgataagct
     3601 gtcaaacatg agaattttct tgaagacgaa agggcctcgt gatacgccta tttttatagg
     3661 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     3721 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     3781 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     3841 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     3901 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     3961 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     4021 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc
     4081 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     4141 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     4201 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     4261 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     4321 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa
     4381 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     4441 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     4501 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     4561 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     4621 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     4681 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     4741 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     4801 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     4861 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     4921 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     4981 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     5041 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     5101 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     5161 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     5221 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     5281 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     5341 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     5401 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     5461 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     5521 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     5581 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     5641 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     5701 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
     5761 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     5821 aatttcacac aggaaacagc tatgaccatg attacgccaa gctctagcta gaggtcgacc
     5881 aattctcatg tttgacagct tatcatcgca gatccgggca acgttgttgc cattgctgca
     5941 ggcgcagaac tggtaggtat ggaagatcta tacattgaat caatattggc aattagccat
     6001 attagtcatt ggttatatag cataaatcaa tattggctat tggccattgc atacgttgta
     6061 tctatatcat aatatgtaca tttatattgg ctcatgtcca atatgaccgc catgttgaca
     6121 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     6181 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     6241 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     6301 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     6361 gtatcatatg ccaagtccgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     6421 ttatgcccag tacatgacct tacgggactt tcctacttgg cagtacatct acgtattagt
     6481 catcgctatt accatg
//
