LOCUS       sequence-275112-        6597 bp    DNA     circular SYN 15-APR-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 6597)
  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 6597)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: D76_Ld_303190
FEATURES             Location/Qualifiers
     source          1..6597
                     /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             1755..1772
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1824..1879
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1878..1897)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4115..4133)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4201..4305
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4306..4374,4375..5166)
                     /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(4524..4543)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5337..5925
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5826..5845
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6079..6096
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6213..6234
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6249..6254,6255..6272,6273..6279)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6287..6303
                     /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     6292..6314
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6311..6327
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6311..6327
                     /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 gcccaagttg
     1201 cgcaggaagc cgagatcaat cctcatccct tgctcttcgt atcaaagacc acatccagcg
     1261 atgatatagt gttggggagc tccgttgaag tggtcgtgac cgttactaac tacggacagt
     1321 cacccgcctt cgatgtgcag atttccgatc tccttgagga tggaagtctg cagagcaaga
     1381 gcattgccta cctgccatac ggagctagtg agaccctgag atataccgtg actccgacag
     1441 cgcttggtaa ttatgctgtg agtgtggcag aggttaccta caacgtcgag cagggtaata
     1501 ctgcgacggc tagaaaggct ctctcaaact tgatccgcga aggagaggct tattattacg
     1561 gtgagggcgt tgacgatgag tcatttcgcg gcgttatcag cgtactcacc agggaccggt
     1621 acgatagact gcacgctcga tatataaagg aatctactgg cgcgcccggc agcctgcacc
     1681 acatcctgga cgcccagaag atgctgtgga accacaggga caggaacctg cctcctctgg
     1741 cccctctggg cccccaccac catcaccacc attgagtcta gacacgtgat taattaagga
     1801 tcccccgacc tcgacctctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg
     1861 gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt tggtcgagat
     1921 ccctcggaga tctctagcta gaggatcgat ccccgccccg gacgaactaa acctgactac
     1981 gacatctctg ccccttcttc gcggggcagt gcatgtaatc ccttcagttg gttggtacaa
     2041 cttgccaact gggccctgtt ccacatgtga cacggggggg gaccaaacac aaaggggttc
     2101 tctgactgta gttgacatcc ttataaatgg atgtgcacat ttgccaacac tgagtggctt
     2161 tcatcctgga gcagactttg cagtctgtgg actgcaacac aacattgcct ttatgtgtaa
     2221 ctcttggctg aagctcttac accaatgctg ggggacatgt acctcccagg ggcccaggaa
     2281 gactacggga ggctacacca acgtcaatca gaggggcctg tgtagctacc gataagcgga
     2341 ccctcaagag ggcattagca atagtgttta taaggccccc ttgttaaccc taaacgggta
     2401 gcatatgctt cccgggtagt agtatatact atccagacta accctaattc aatagcatat
     2461 gttacccaac gggaagcata tgctatcgaa ttagggttag taaaagggtc ctaaggaaca
     2521 gcgatatctc ccaccccatg agctgtcacg gttttattta catggggtca ggattccacg
     2581 agggtagtga accattttag tcacaagggc agtggctgaa gatcaaggag cgggcagtga
     2641 actctcctga atcttcgcct gcttcttcat tctccttcgt ttagctaata gaataactgc
     2701 tgagttgtga acagtaaggt gtatgtgagg tgctcgaaaa caaggtttca ggtgacgccc
     2761 ccagaataaa atttggacgg ggggttcagt ggtggcattg tgctatgaca ccaatataac
     2821 cctcacaaac cccttgggca ataaatacta gtgtaggaat gaaacattct gaatatcttt
     2881 aacaatagaa atccatgggg tggggacaag ccgtaaagac tggatgtcca tctcacacga
     2941 atttatggct atgggcaaca cataatccta gtgcaatatg atactggggt tattaagatg
     3001 tgtcccaggc agggaccaag acaggtgaac catgttgtta cactctattt gtaacaaggg
     3061 gaaagagagt ggacgccgac agcagcggac tccactggtt gtctctaaca cccccgaaaa
     3121 ttaaacgggg ctccacgcca atggggccca taaacaaaga caagtggcca ctcttttttt
     3181 tgaaattgtg gagtgggggc acgcgtcagc ccccacacgc cgccctgcgg ttttggactg
     3241 taaaataagg gtgtaataac ttggctgatt gtaaccccgc taaccactgc ggtcaaacca
     3301 cttgcccaca aaaccactaa tggcaccccg gggaatacct gcataagtag gtgggcgggc
     3361 caagataggg gcgcgattgc tgcgatctgg aggacaaatt acacacactt gcgcctgagc
     3421 gccaagcaca gggttgttgg tcctcatatt cacgaggtcg ctgagagcac ggtgggctaa
     3481 tgttgccatg ggtagcatat actacccaaa tatctggata gcatatgcta tcctaatcta
     3541 tatctgggta gcataggcta tcctaatcta tatctgggta gcatatgcta tcctaatcta
     3601 tatctgggta gtatatgcta tcctaattta tatctgggta gcataggcta tcctaatcta
     3661 tatctgggta gcatatgcta tcctaatcta tatctgggta gtatatgcta tcctaatctg
     3721 tatccgggta gcatatgcta tcctaataga gattagggta gtatatgcta tcctaattta
     3781 tatctgggta gcatatacta cccaaatatc tggatagcat atgctatcct aatctatatc
     3841 tgggtagcat atgctatcct aatctatatc tgggtagcat aggctatcct aatctatatc
     3901 tgggtagcat atgctatcct aatctatatc tgggtagtat atgctatcct aatttatatc
     3961 tgggtagcat aggctatcct aatctatatc tgggtagcat atgctatcct aatctatatc
     4021 tgggtagtat atgctatcct aatctgtatc cgggtagcat atgctatcct catgataagc
     4081 tgtcaaacat gagaattttc ttgaagacga aagggcctcg tgatacgcct atttttatag
     4141 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     4201 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     4261 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     4321 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     4381 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     4441 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     4501 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg
     4561 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     4621 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     4681 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     4741 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     4801 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca
     4861 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     4921 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     4981 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     5041 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     5101 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     5161 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     5221 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     5281 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5341 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5401 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     5461 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     5521 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     5581 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     5641 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     5701 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     5761 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     5821 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     5881 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     5941 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     6001 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     6061 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     6121 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     6181 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     6241 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     6301 caatttcaca caggaaacag ctatgaccat gattacgcca agctctagct agaggtcgac
     6361 caattctcat gtttgacagc ttatcatcgc agatccgggc aacgttgttg ccattgctgc
     6421 aggcgcagaa ctggtaggta tggaagatct atacattgaa tcaatattgg caattagcca
     6481 tattagtcat tggttatata gcataaatca atattggcta ttggccattg catacgttgt
     6541 atctatatca taatatgtac atttatattg gctcatgtcc aatatgaccg ccatgtt
//
