LOCUS       sequence-274511-        8376 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 8376)
  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 8376)
  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: D4__Ld_290790
FEATURES             Location/Qualifiers
     source          1..8376
                     /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             3534..3551
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3603..3658
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3657..3676)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5894..5912)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5980..6084
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6085..6153,6154..6945)
                     /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(6303..6322)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7116..7704
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7605..7624
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7858..7875
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7992..8013
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8028..8033,8034..8051,8052..8058)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8066..8082
                     /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     8071..8093
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8090..8106
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8090..8106
                     /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 gccaccgggg
     1201 atggcagagg caccccaatc gcattccagg ccgaggtgtc caagatgctg gacatcctgg
     1261 tcaacagcct gtataccaac cgggccgtgt tcctgagaga gctgatcagc aacggcagcg
     1321 acgccctgga caagatccgg gtgctgtacc tgaccagccc caaagagccc ctgaccaagg
     1381 acggcgaggc ccccaccatg gacctgcgga tcagcttcga caaagagaag tccgagctga
     1441 tcctgcggga cggcggcgtg ggcatgacca aagaggaact ggccaagcac ctgggcagcc
     1501 tgggcaccag cggcaccaag cactttctgg aaaagctgca ggaaggcgtc ggcgccgtcg
     1561 gaggcgacca gaacaatctg atcggccagt tcggcgtggg cttctacagc gtgttcctcg
     1621 tgggcgacag agtgcgggtg gccagcaaga gcgacgacag cgacgagcag tacgtgtggg
     1681 agagcaaggg cgacggccag tacttcctgt accccgaccc cagaggcaac accctgggca
     1741 ggggcaccga gatcaccatc gagctgaagc ccgacgccga gcagttcctg agcgccgaga
     1801 caatcaagaa aaccatccac cagtacagcg agttcatcaa cttccccatc tacgtgcagg
     1861 aagaggtcga ggtcgccagc accgccgcca cacctgagtc tgccgccgag gaaagaagcc
     1921 tggacgaggg cgccgtggaa gaggaccccg acaaagaggg cgacacccag ggcgtggtca
     1981 aagaaaagcg gtggaccctg gtcaacgaga accggcccat ctggacccgg cccatcggca
     2041 acgtgaccga ggaagagtac cacaagttct acaaggcctt cagcggcgac taccgggacc
     2101 ccctgtactt caaccacttc aaggtggaag gcgaggtgga cttcgacagc atcctgttcg
     2161 tgcccaccac cgtggacccc gccagcttca gcgacgacaa cgccgtgccc aacaccaaca
     2221 tcaagctgta cgtgcggcgg gtgttcatca ccgacgagtt ccgggacctg ctgcccagat
     2281 acctgaactt cgtgaagggc atcgtggaca gcaacgacct gcccctgaac gtgtccagag
     2341 aggtgctgca ggaaagccgg atcctgagag tgatcaagaa aaagctcgtg cgcaagaccc
     2401 tgagcatgtt cgccgatatc gccgcccagg acgaggccat tgccaacggc aaacaggtgg
     2461 aaaaccctgc ccccagcggc cacacccacc tgaagaagcc cgcctacacc aagttctggg
     2521 agctgtacgg caagcacctg agactgggcg tgatgctgga ctccaacaac cggaaccggc
     2581 tgaccaagct gttccggtac aagagcagca gaagcgagag cgagtacatc agcctgcaga
     2641 cctacgtgga ccggatgaag aagggccaga agggcatcta ctacctgagc ggcgacagcg
     2701 tggcccggat caagaaatcc cccgtgctgg aagatgccgt gaaccacgac gtggaagtga
     2761 tcttcatgac cgacgccatc gacgagtacg tggtgtccca gctgaccgac ttcgccggca
     2821 agaagctgat caacctggcc aaagaaggcg tgcagttcga ggaaagcgac gcccggcagc
     2881 gggtggtgga ccggaagcgg aaagagaagt acgacagctt ctttacccac ctgagagccc
     2941 tgttcggcta cagcgaagtg cggaaagtga tcctgaccaa gcggatgacc aacgaggcct
     3001 tcatcgtgtc cagcagcgag aaccagatca ccgccagact ggccagcatc atgcggggcc
     3061 agagcatgag cctggccaac cagcagatga ccgccgagcg ggtgctggaa gtgaactaca
     3121 gacaccccct ggtggacgag atgttcaagc ggttcaccgt ggacgaggac gacgaggtgg
     3181 ccaccgacat tgcctgggtg ctgtatgaca ccgccaacct gcaggccgag ttccccgtgg
     3241 ccgatgtggc cgcctacagc aagcggatca accggctgct gcggagcagc gtggacctgt
     3301 ccgccgacga tagcctgctg ccccccgacg atgccgagta caccgtgtcc gacaccgaaa
     3361 ccgaagagga agaggaacag cccaaggtgg acaccaacgc ccacgaagag gccgagacag
     3421 acggcgaggg agacctgggc gcgcccggca gcctgcacca catcctggac gcccagaaga
     3481 tgctgtggaa ccacagggac aggaacctgc ctcctctggc ccctctgggc ccccaccacc
     3541 atcaccacca ttgagtctag acacgtgatt aattaaggat cccccgacct cgacctctgg
     3601 ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact
     3661 cggaaggaca tatgggaggg caaatcattt ggtcgagatc cctcggagat ctctagctag
     3721 aggatcgatc cccgccccgg acgaactaaa cctgactacg acatctctgc cccttcttcg
     3781 cggggcagtg catgtaatcc cttcagttgg ttggtacaac ttgccaactg ggccctgttc
     3841 cacatgtgac acgggggggg accaaacaca aaggggttct ctgactgtag ttgacatcct
     3901 tataaatgga tgtgcacatt tgccaacact gagtggcttt catcctggag cagactttgc
     3961 agtctgtgga ctgcaacaca acattgcctt tatgtgtaac tcttggctga agctcttaca
     4021 ccaatgctgg gggacatgta cctcccaggg gcccaggaag actacgggag gctacaccaa
     4081 cgtcaatcag aggggcctgt gtagctaccg ataagcggac cctcaagagg gcattagcaa
     4141 tagtgtttat aaggccccct tgttaaccct aaacgggtag catatgcttc ccgggtagta
     4201 gtatatacta tccagactaa ccctaattca atagcatatg ttacccaacg ggaagcatat
     4261 gctatcgaat tagggttagt aaaagggtcc taaggaacag cgatatctcc caccccatga
     4321 gctgtcacgg ttttatttac atggggtcag gattccacga gggtagtgaa ccattttagt
     4381 cacaagggca gtggctgaag atcaaggagc gggcagtgaa ctctcctgaa tcttcgcctg
     4441 cttcttcatt ctccttcgtt tagctaatag aataactgct gagttgtgaa cagtaaggtg
     4501 tatgtgaggt gctcgaaaac aaggtttcag gtgacgcccc cagaataaaa tttggacggg
     4561 gggttcagtg gtggcattgt gctatgacac caatataacc ctcacaaacc ccttgggcaa
     4621 taaatactag tgtaggaatg aaacattctg aatatcttta acaatagaaa tccatggggt
     4681 ggggacaagc cgtaaagact ggatgtccat ctcacacgaa tttatggcta tgggcaacac
     4741 ataatcctag tgcaatatga tactggggtt attaagatgt gtcccaggca gggaccaaga
     4801 caggtgaacc atgttgttac actctatttg taacaagggg aaagagagtg gacgccgaca
     4861 gcagcggact ccactggttg tctctaacac ccccgaaaat taaacggggc tccacgccaa
     4921 tggggcccat aaacaaagac aagtggccac tctttttttt gaaattgtgg agtgggggca
     4981 cgcgtcagcc cccacacgcc gccctgcggt tttggactgt aaaataaggg tgtaataact
     5041 tggctgattg taaccccgct aaccactgcg gtcaaaccac ttgcccacaa aaccactaat
     5101 ggcaccccgg ggaatacctg cataagtagg tgggcgggcc aagatagggg cgcgattgct
     5161 gcgatctgga ggacaaatta cacacacttg cgcctgagcg ccaagcacag ggttgttggt
     5221 cctcatattc acgaggtcgc tgagagcacg gtgggctaat gttgccatgg gtagcatata
     5281 ctacccaaat atctggatag catatgctat cctaatctat atctgggtag cataggctat
     5341 cctaatctat atctgggtag catatgctat cctaatctat atctgggtag tatatgctat
     5401 cctaatttat atctgggtag cataggctat cctaatctat atctgggtag catatgctat
     5461 cctaatctat atctgggtag tatatgctat cctaatctgt atccgggtag catatgctat
     5521 cctaatagag attagggtag tatatgctat cctaatttat atctgggtag catatactac
     5581 ccaaatatct ggatagcata tgctatccta atctatatct gggtagcata tgctatccta
     5641 atctatatct gggtagcata ggctatccta atctatatct gggtagcata tgctatccta
     5701 atctatatct gggtagtata tgctatccta atttatatct gggtagcata ggctatccta
     5761 atctatatct gggtagcata tgctatccta atctatatct gggtagtata tgctatccta
     5821 atctgtatcc gggtagcata tgctatcctc atgataagct gtcaaacatg agaattttct
     5881 tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     5941 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     6001 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     6061 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     6121 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     6181 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     6241 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     6301 ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc
     6361 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     6421 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     6481 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     6541 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     6601 aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta
     6661 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     6721 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     6781 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     6841 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     6901 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     6961 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     7021 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     7081 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     7141 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     7201 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     7261 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     7321 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     7381 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     7441 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     7501 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     7561 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     7621 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     7681 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     7741 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     7801 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     7861 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     7921 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     7981 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     8041 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     8101 tatgaccatg attacgccaa gctctagcta gaggtcgacc aattctcatg tttgacagct
     8161 tatcatcgca gatccgggca acgttgttgc cattgctgca ggcgcagaac tggtaggtat
     8221 ggaagatcta tacattgaat caatattggc aattagccat attagtcatt ggttatatag
     8281 cataaatcaa tattggctat tggccattgc atacgttgta tctatatcat aatatgtaca
     8341 tttatattgg ctcatgtcca atatgaccgc catgtt
//
