LOCUS       sequence-274549-        8643 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 8643)
  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 8643)
  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: D23_Ld_170240
FEATURES             Location/Qualifiers
     source          1..8643
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     1086..1105
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1128..1137
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3801..3818
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3870..3925
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3924..3943)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6161..6179)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6247..6351
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6352..6420,6421..7212)
                     /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(6570..6589)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7383..7971
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7872..7891
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8125..8142
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8259..8280
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8295..8300,8301..8318,8319..8325)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8333..8349
                     /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     8338..8360
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8357..8373
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8357..8373
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agatcctcac tctcttccgc atcgctgtct gcgagggcca gctgttgggc tcgcggttga
      661 ggacaaactc ttcgcggtct ttccagtact cttggatcgg aaacccgtcg gcctccgaac
      721 ggtactccgc caccgaggga cctgagcgag tccgcatcga ccggatcgga aaacctctcg
      781 agaaaggcgt ctaaccagtc acagtcgcaa ggtaggctga gcaccgtggc gggcggcagc
      841 gggtggcggt cggggttgtt tctggcggag gtgctgctga tgatgtaatt aaagtaggcg
      901 gtcttgagac ggcggatggt cgaggtgagg tgtggcaggc ttgagatcca gctgttgggg
      961 tgagtactcc ctctcaaaag cgggcattac ttctgcgcta agattgtcag tttccaaaaa
     1021 cgaggaggat ttgatattca cctggcccga tctggccata cacttgagtg acaatgacat
     1081 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caagtttgcc accatggagt
     1141 ttcagaccca ggtactcatg tccctgctgc tctgcatgtc tggtgcggcc gccacctttg
     1201 atggcgacgc cgtgacagcc cctcctagcg ccagcaccgc cttcaccgag caccctttca
     1261 ccctgcctac cgccgccagc ctgctggacg tgctggccag cgacttctgc accagaaccg
     1321 gcgagggcga ggaaagcagc tgcaccccca gcgtgtccag actgcccacc caccccgtgc
     1381 tgatcctgtt taccgagagc aacgccggcc acagcagcac cctggccgcc ctgagagcca
     1441 aagtgcggca gatcgccacc gacgtgccca gcagcgtgct gcgggtgcac gagtacgtga
     1501 tccccatgag cgccacagag cggcggctga tcgagctgct gctgatcggc gccagccacg
     1561 tgccaacact ggccctgttt cacggccggg tggccaaggt gcaggtcgcc cctggatcca
     1621 tcgtgtctgc cagccccttc cagacccctg tgctgtactc caccaccccc ctggcctccg
     1681 ccagctacct ggaactgaga agctgggccc tgtccgagct gcccgccaga tacgtggacc
     1741 ccgtgacctt ccacctgatc cctagcctgc agttcgtgtt cagccctgcc gaagtgcacg
     1801 aaacactgca gctcgtgcgg gacgccctgg aagccagcga gagcagatcc gtgctgcctg
     1861 ccttcgtgtc catggcctac atgcggctga cccggcacgg cagcgaagag gtgctggctg
     1921 ccctgtctag cattgccacc caggccggca acgctgtgct gaccctggtc accgaaagcg
     1981 ccgaggtggc cgctgcctgg ggactgagag ctgagcacac cttcaccatg gccccttggt
     2041 ccagcacagt ggccgcctac ctgaacagca gcgcctccga tggcttcgcc gcctccgtgt
     2101 ccgatctgag ccctccccag agcatcggca ccgtgaccga gctggccgaa gccacagccg
     2161 gctctgccgc ttggagagcc gcttgtgccg cctctgtgcg ggccgagatc gcccagctgc
     2221 agaactggcg gcaggccatg aacgccttca acaccaccag ccctctgcgg cggatcgaca
     2281 gcgccgctca tctgctgcac gagctgaagg ccctgcagaa tgccctgaag atcgtgttcg
     2341 tgctgagaga aagcgacgag atgtggtttc accaccacct ggaagtggct gccgctctgg
     2401 ccagaaggct gcagagcacc accgtgttct acaacacaac cacactggct aagggcagca
     2461 aggcccggtc ccgggtggaa agatcctgga cccccatcat gagatgcgac gtgttctggc
     2521 tggacgccga gcagctccct gccgtggctg atggactgca cgtgtcccag gtgcccagtg
     2581 tgctgattct ggtgcccctg cagtccagat tcggagaggg cggcgacggc gatagagccg
     2641 atgctgctgc agccgctggc gacgacgatc aggcagctgc cggcgcttct gaggaagccg
     2701 gcctgagaag gcgggacccc ttcatcggca tccacaccat caacagatac gacctgctga
     2761 aagccgccta tatggacgac gcccagatga cccacgagcc tgtgtccggc aaagaggcac
     2821 tgcctctgtt ccccagcgat tccgatgccc tgctgcggtt cctggctagc ggcagctttc
     2881 tgggcgccat gcagtacacc atgtcctcca cccagctgtc taccctgcgg gctaaagtga
     2941 cagagctgcc tgatgccgtg caggccgcta ccgccagcgg cggaaccagc aacaggcact
     3001 acctgcagct ggaccggcgg tactaccctc tgagaggggc cgaggaaccc atggaaggcc
     3061 ccgtgtacgt cagacagatc ctgaacggct ccagccccct gccagtgctg ggctccgacg
     3121 acagcgacac cgaactgggc agacagcccg tgggcggcag aagcggcaca caccctagag
     3181 gcaccacctc cgcctccccc gacgaggccg atgagagact gagacagcgg agagccagag
     3241 aagaggccgc agccctgaaa cggaaggccg cctgggaggc caatctggcc aagcggagaa
     3301 gggaaaaggc cgaccggatg cggagaaagg ctgccgagga tgccgccgag cgcgagaggc
     3361 agcaggaagc cttccagcag gaagtgaacg ccgccctgaa ggccgaacag gaaggcgctg
     3421 gcggcggagc tggcggaaca tgggcttctc agcctgaggg cctgctcgtc agaaggccca
     3481 ctctggaaga tgccagcgaa ggcctggtgg aagagggcag acggaagggc aggccccaga
     3541 ggcccttcgg ggaggaagtg gatagccagg acgctgccca gattctccgg cggaggcgga
     3601 gacaccgcga gtacagagag tggctggctg acagacagct gatggtcaat agatgcgtga
     3661 cagtgaccga aaagggcaag ctgagcctga tgaccagatg ggaaggcgcg cccggcagcc
     3721 tgcaccacat cctggacgcc cagaagatgc tgtggaacca cagggacagg aacctgcctc
     3781 ctctggcccc tctgggcccc caccaccatc accaccattg agtctagaca cgtgattaat
     3841 taaggatccc ccgacctcga cctctggcta ataaaggaaa tttattttca ttgcaatagt
     3901 gtgttggaat tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttggt
     3961 cgagatccct cggagatctc tagctagagg atcgatcccc gccccggacg aactaaacct
     4021 gactacgaca tctctgcccc ttcttcgcgg ggcagtgcat gtaatccctt cagttggttg
     4081 gtacaacttg ccaactgggc cctgttccac atgtgacacg gggggggacc aaacacaaag
     4141 gggttctctg actgtagttg acatccttat aaatggatgt gcacatttgc caacactgag
     4201 tggctttcat cctggagcag actttgcagt ctgtggactg caacacaaca ttgcctttat
     4261 gtgtaactct tggctgaagc tcttacacca atgctggggg acatgtacct cccaggggcc
     4321 caggaagact acgggaggct acaccaacgt caatcagagg ggcctgtgta gctaccgata
     4381 agcggaccct caagagggca ttagcaatag tgtttataag gcccccttgt taaccctaaa
     4441 cgggtagcat atgcttcccg ggtagtagta tatactatcc agactaaccc taattcaata
     4501 gcatatgtta cccaacggga agcatatgct atcgaattag ggttagtaaa agggtcctaa
     4561 ggaacagcga tatctcccac cccatgagct gtcacggttt tatttacatg gggtcaggat
     4621 tccacgaggg tagtgaacca ttttagtcac aagggcagtg gctgaagatc aaggagcggg
     4681 cagtgaactc tcctgaatct tcgcctgctt cttcattctc cttcgtttag ctaatagaat
     4741 aactgctgag ttgtgaacag taaggtgtat gtgaggtgct cgaaaacaag gtttcaggtg
     4801 acgcccccag aataaaattt ggacgggggg ttcagtggtg gcattgtgct atgacaccaa
     4861 tataaccctc acaaacccct tgggcaataa atactagtgt aggaatgaaa cattctgaat
     4921 atctttaaca atagaaatcc atggggtggg gacaagccgt aaagactgga tgtccatctc
     4981 acacgaattt atggctatgg gcaacacata atcctagtgc aatatgatac tggggttatt
     5041 aagatgtgtc ccaggcaggg accaagacag gtgaaccatg ttgttacact ctatttgtaa
     5101 caaggggaaa gagagtggac gccgacagca gcggactcca ctggttgtct ctaacacccc
     5161 cgaaaattaa acggggctcc acgccaatgg ggcccataaa caaagacaag tggccactct
     5221 tttttttgaa attgtggagt gggggcacgc gtcagccccc acacgccgcc ctgcggtttt
     5281 ggactgtaaa ataagggtgt aataacttgg ctgattgtaa ccccgctaac cactgcggtc
     5341 aaaccacttg cccacaaaac cactaatggc accccgggga atacctgcat aagtaggtgg
     5401 gcgggccaag ataggggcgc gattgctgcg atctggagga caaattacac acacttgcgc
     5461 ctgagcgcca agcacagggt tgttggtcct catattcacg aggtcgctga gagcacggtg
     5521 ggctaatgtt gccatgggta gcatatacta cccaaatatc tggatagcat atgctatcct
     5581 aatctatatc tgggtagcat aggctatcct aatctatatc tgggtagcat atgctatcct
     5641 aatctatatc tgggtagtat atgctatcct aatttatatc tgggtagcat aggctatcct
     5701 aatctatatc tgggtagcat atgctatcct aatctatatc tgggtagtat atgctatcct
     5761 aatctgtatc cgggtagcat atgctatcct aatagagatt agggtagtat atgctatcct
     5821 aatttatatc tgggtagcat atactaccca aatatctgga tagcatatgc tatcctaatc
     5881 tatatctggg tagcatatgc tatcctaatc tatatctggg tagcataggc tatcctaatc
     5941 tatatctggg tagcatatgc tatcctaatc tatatctggg tagtatatgc tatcctaatt
     6001 tatatctggg tagcataggc tatcctaatc tatatctggg tagcatatgc tatcctaatc
     6061 tatatctggg tagtatatgc tatcctaatc tgtatccggg tagcatatgc tatcctcatg
     6121 ataagctgtc aaacatgaga attttcttga agacgaaagg gcctcgtgat acgcctattt
     6181 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     6241 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     6301 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     6361 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     6421 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     6481 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     6541 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac
     6601 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     6661 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     6721 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     6781 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     6841 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca
     6901 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     6961 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     7021 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     7081 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     7141 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     7201 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     7261 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     7321 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     7381 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     7441 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     7501 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     7561 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     7621 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     7681 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     7741 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     7801 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     7861 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     7921 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     7981 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     8041 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     8101 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     8161 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     8221 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     8281 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     8341 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct ctagctagag
     8401 gtcgaccaat tctcatgttt gacagcttat catcgcagat ccgggcaacg ttgttgccat
     8461 tgctgcaggc gcagaactgg taggtatgga agatctatac attgaatcaa tattggcaat
     8521 tagccatatt agtcattggt tatatagcat aaatcaatat tggctattgg ccattgcata
     8581 cgttgtatct atatcataat atgtacattt atattggctc atgtccaata tgaccgccat
     8641 gtt
//
