LOCUS       sequence-274532-        8424 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 8424)
  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 8424)
  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: D16_Ld_090240
FEATURES             Location/Qualifiers
     source          1..8424
                     /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             3202..3219
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3271..3326
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3325..3344)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5562..5580)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5648..5752
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5753..5821,5822..6613)
                     /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(5971..5990)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6784..7372
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7273..7292
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7526..7543
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7660..7681
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7696..7701,7702..7719,7720..7726)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7734..7750
                     /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     7739..7761
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7758..7774
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7758..7774
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        8045..8424
                     /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 gccacctact tcacctttct gaacgtgccc
      841 gtggaagccg aggaacccgg cagcacagct acagcctctg ctggcgacgg cgtgtccgcc
      901 atggtcggag ctgctccatc tggcgccctg cctggcacag gatccgatgg cgccgagtac
      961 ggccacgcct acgaccctct gctcctgctg cctcacgagc tgcccctgta caccagcatc
     1021 accagatacg tgtccctggc cgaggacacc accctgggaa tcgcctaccg gtggctgctg
     1081 atcaacggcc tgcactacag ccccagagag gtgtacggcg agcggcggct gagagagtgg
     1141 ggctactgct acagcagcag cgtgggcgga ccctccggcg gcaaagatgt ggatgtggcc
     1201 ctgagcggag ccggcatgag agtgtccgac agacctagca gcgccgtggc cgcctctctg
     1261 tctagcgccg gatctgccat ggccgccagc ctggacagca gccggtgcag cagaagaagg
     1321 ctgagaagag gcgccgctgc cgctgctgcc gcacctccta cacccctgag aagggaaagc
     1381 gcccactggg tcaacgtgct gattagatgc gccgccttcc tgtgcctggg cggaggcacc
     1441 gtgaagcccg aagtgtggac cgaccatctg ctgtacgggg ccgagaaggt gctggaaagc
     1501 gtgaacgtgg gctacgagaa ccggatgcgg gccagagccg cccaggctca tgctgtggca
     1561 gcccagctgg cctctgccgc ccatcctagc cctccacctc acagagccag cagccagagc
     1621 gctgccccac tgctgaccgc ccctagaaag cccctgctgc ggatccagct gctggaactg
     1681 ggctctggcc tgctgggcgg cacactgaga gatgtgcgca gaccccccct gagcctgcac
     1741 catggcgcct ctgatgccag cggcagctct gccacccctg ccagagctac ccctcctgct
     1801 agaagggccg ctaccgcagc cgccgctgga tctcctggaa tggccccatc tgccgcaggc
     1861 ttcgcccacc atcaccacca cagcaacagc tccgtgtcca gcgaccacag acagcagatc
     1921 agctcccacg ccgccctgta cgccccaagc agcctgctgt cctctagcgt ggccggcctg
     1981 agcaccgcca tcaccgatgc cgtgtcctcc gtgaccacca ggggcaccga ttccgcccac
     2041 ctgagcgatc ctgtggctgg cgctccagtg gccgattacg ccggctcctg ccctctgttt
     2101 actggcggcg gagggggagc ctcttctgcc ggcgctcagc tgggagtggt gctgcccaga
     2161 gtggaaggcg acgtgatcag cgtggaacag gcctacagcg agggctgcca catgaggcct
     2221 ccaaccccag ccgcctccaa tagcaccccc gctagagccc tgtctgcagc cgctagcgcc
     2281 agccctctgc ctagagccag tgccgccttt cagctgcggt gcttcgccgt gcctctcctg
     2341 tacgaggacc agcggtttca cgtgcggctg ggctgctgtc tgcctctggg agcactgctg
     2401 cccgtgtctc tgtgcgtgcc ccccgatgtg ctgaccctga actgcgctgt ggccgtgcac
     2461 agagtgacct tcaccggcca cctgtatgcc gctttccacg gcgccagggt ggaactgagc
     2521 ttccccacag cccccctgtt caccgccgtg gtcaaagtga tgcccgacac cagcgacgcc
     2581 agctccggca gcggaagcgt ggcagaagtg tcccatcacg gcggatgggg catgagggca
     2641 gctgggggag gcggagtgca tagcggagca gccgcagcag ctgccactac cgcctttacc
     2701 gccacataca cccagcggaa cgaggccacc gtcgtgggag gcagcagcat cgatggcggc
     2761 agagactcct acaacgccgc agcccgacct cccctgcctc ccagaagcgg cctgtacagc
     2821 tctggctcca atgcctccct gaaccaccac catagatccg gcatgtccgg gggagggggc
     2881 acaagcgcct ccggctcttg cctgaacgag cacaacgaga aagtgcagga agtggtcctg
     2941 ctggctgtgc ggagactgat cgagagcctg acctacccca aggtgctcgt gggcgaactc
     3001 gtgtgcagaa ggcctagcgg cgctggcgga acagccgagg gcggagctga aatgcagctg
     3061 cagctcagat ggaccagaca gaccgctagc ctgccactgt gcatgggcgc gcccggcagc
     3121 ctgcaccaca tcctggacgc ccagaagatg ctgtggaacc acagggacag gaacctgcct
     3181 cctctggccc ctctgggccc ccaccaccat caccaccatt gagtctagac acgtgattaa
     3241 ttaaggatcc cccgacctcg acctctggct aataaaggaa atttattttc attgcaatag
     3301 tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttgg
     3361 tcgagatccc tcggagatct ctagctagag gatcgatccc cgccccggac gaactaaacc
     3421 tgactacgac atctctgccc cttcttcgcg gggcagtgca tgtaatccct tcagttggtt
     3481 ggtacaactt gccaactggg ccctgttcca catgtgacac ggggggggac caaacacaaa
     3541 ggggttctct gactgtagtt gacatcctta taaatggatg tgcacatttg ccaacactga
     3601 gtggctttca tcctggagca gactttgcag tctgtggact gcaacacaac attgccttta
     3661 tgtgtaactc ttggctgaag ctcttacacc aatgctgggg gacatgtacc tcccaggggc
     3721 ccaggaagac tacgggaggc tacaccaacg tcaatcagag gggcctgtgt agctaccgat
     3781 aagcggaccc tcaagagggc attagcaata gtgtttataa ggcccccttg ttaaccctaa
     3841 acgggtagca tatgcttccc gggtagtagt atatactatc cagactaacc ctaattcaat
     3901 agcatatgtt acccaacggg aagcatatgc tatcgaatta gggttagtaa aagggtccta
     3961 aggaacagcg atatctccca ccccatgagc tgtcacggtt ttatttacat ggggtcagga
     4021 ttccacgagg gtagtgaacc attttagtca caagggcagt ggctgaagat caaggagcgg
     4081 gcagtgaact ctcctgaatc ttcgcctgct tcttcattct ccttcgttta gctaatagaa
     4141 taactgctga gttgtgaaca gtaaggtgta tgtgaggtgc tcgaaaacaa ggtttcaggt
     4201 gacgccccca gaataaaatt tggacggggg gttcagtggt ggcattgtgc tatgacacca
     4261 atataaccct cacaaacccc ttgggcaata aatactagtg taggaatgaa acattctgaa
     4321 tatctttaac aatagaaatc catggggtgg ggacaagccg taaagactgg atgtccatct
     4381 cacacgaatt tatggctatg ggcaacacat aatcctagtg caatatgata ctggggttat
     4441 taagatgtgt cccaggcagg gaccaagaca ggtgaaccat gttgttacac tctatttgta
     4501 acaaggggaa agagagtgga cgccgacagc agcggactcc actggttgtc tctaacaccc
     4561 ccgaaaatta aacggggctc cacgccaatg gggcccataa acaaagacaa gtggccactc
     4621 ttttttttga aattgtggag tgggggcacg cgtcagcccc cacacgccgc cctgcggttt
     4681 tggactgtaa aataagggtg taataacttg gctgattgta accccgctaa ccactgcggt
     4741 caaaccactt gcccacaaaa ccactaatgg caccccgggg aatacctgca taagtaggtg
     4801 ggcgggccaa gataggggcg cgattgctgc gatctggagg acaaattaca cacacttgcg
     4861 cctgagcgcc aagcacaggg ttgttggtcc tcatattcac gaggtcgctg agagcacggt
     4921 gggctaatgt tgccatgggt agcatatact acccaaatat ctggatagca tatgctatcc
     4981 taatctatat ctgggtagca taggctatcc taatctatat ctgggtagca tatgctatcc
     5041 taatctatat ctgggtagta tatgctatcc taatttatat ctgggtagca taggctatcc
     5101 taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagta tatgctatcc
     5161 taatctgtat ccgggtagca tatgctatcc taatagagat tagggtagta tatgctatcc
     5221 taatttatat ctgggtagca tatactaccc aaatatctgg atagcatatg ctatcctaat
     5281 ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagcatagg ctatcctaat
     5341 ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat
     5401 ttatatctgg gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat
     5461 ctatatctgg gtagtatatg ctatcctaat ctgtatccgg gtagcatatg ctatcctcat
     5521 gataagctgt caaacatgag aattttcttg aagacgaaag ggcctcgtga tacgcctatt
     5581 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     5641 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     5701 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     5761 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     5821 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     5881 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     5941 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga
     6001 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     6061 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     6121 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     6181 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     6241 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc
     6301 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     6361 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     6421 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     6481 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     6541 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     6601 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     6661 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     6721 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     6781 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     6841 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     6901 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     6961 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     7021 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     7081 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     7141 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     7201 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     7261 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     7321 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     7381 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     7441 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     7501 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     7561 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     7621 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     7681 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     7741 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc tctagctaga
     7801 ggtcgaccaa ttctcatgtt tgacagctta tcatcgcaga tccgggcaac gttgttgcca
     7861 ttgctgcagg cgcagaactg gtaggtatgg aagatctata cattgaatca atattggcaa
     7921 ttagccatat tagtcattgg ttatatagca taaatcaata ttggctattg gccattgcat
     7981 acgttgtatc tatatcataa tatgtacatt tatattggct catgtccaat atgaccgcca
     8041 tgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
     8101 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
     8161 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
     8221 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     8281 catcaagtgt atcatatgcc aagtccgccc cctattgacg tcaatgacgg taaatggccc
     8341 gcctggcatt atgcccagta catgacctta cgggactttc ctacttggca gtacatctac
     8401 gtattagtca tcgctattac catg
//
