LOCUS       sequence-262342-        8145 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 8145)
  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 8145)
  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: D50_Ld_100670
FEATURES             Location/Qualifiers
     source          1..8145
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        7..386
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        387..590
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     541..561
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     587..611
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     1092..1111
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1134..1143
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3309..3326
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3378..3433
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3432..3451)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5669..5687)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5755..5859
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5860..5928,5929..6720)
                     /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(6078..6097)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6891..7479
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7380..7399
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7633..7650
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7767..7788
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7803..7808,7809..7826,7827..7833)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7841..7857
                     /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     7846..7868
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7865..7881
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7865..7881
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 catgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
       61 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      121 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      181 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      241 tacatcaagt gtatcatatg ccaagtccgc cccctattga cgtcaatgac ggtaaatggc
      301 ccgcctggca ttatgcccag tacatgacct tacgggactt tcctacttgg cagtacatct
      361 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacacc aatgggcgtg
      421 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      481 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taataacccc gccccgttga
      541 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga
      601 accgtcagat cctcactctc ttccgcatcg ctgtctgcga gggccagctg ttgggctcgc
      661 ggttgaggac aaactcttcg cggtctttcc agtactcttg gatcggaaac ccgtcggcct
      721 ccgaacggta ctccgccacc gagggacctg agcgagtccg catcgaccgg atcggaaaac
      781 ctctcgagaa aggcgtctaa ccagtcacag tcgcaaggta ggctgagcac cgtggcgggc
      841 ggcagcgggt ggcggtcggg gttgtttctg gcggaggtgc tgctgatgat gtaattaaag
      901 taggcggtct tgagacggcg gatggtcgag gtgaggtgtg gcaggcttga gatccagctg
      961 ttggggtgag tactccctct caaaagcggg cattacttct gcgctaagat tgtcagtttc
     1021 caaaaacgag gaggatttga tattcacctg gcccgatctg gccatacact tgagtgacaa
     1081 tgacatccac tttgcctttc tctccacagg tgtccactcc caggtccaag tttgccacca
     1141 tggagtttca gacccaggta ctcatgtccc tgctgctctg catgtctggt gcggccgcca
     1201 ccgctgccgt ggccacaaca acagccagag cctacgacca cgccggcatc acagtggctg
     1261 gcgccctgct cgtgggccag aacctgcagg gaaaggcagg cgccagccgg atcctgaacc
     1321 ccttcgccat ctgcgccgac ttcgacaccg ccgacgtgga agataccacc ctgctgatcg
     1381 gcggagccag ctacttcttc agcttcaacc ggtacagcac ctacctgggc ttttggtacg
     1441 gccagggcag catgaacctg aacagcggcc ccatcgacaa agtgcggctg accggcgtgt
     1501 tcggctgcgt gaccctgcgg cccaacagca gcaatggcct ggtcaccagc accgtgtact
     1561 acgtgcagaa cgacggcatg ctgtactggg tgtccaacag cgtggtgtac ctgacccagg
     1621 tcaaacacgg catcagcttc gtggacgtga ccgtgcacga caacaatgtg tatctgctga
     1681 gcacccagaa cggcatctac cagtgcggca tcggcgctgg cggagctgtc gtgggcagcg
     1741 cctgtaccca gatcaccctg acaggcagca ccgagttcca ccagctgatc cccatcagca
     1801 gcgacttccg gggcttcgcc gtgtccagct ccggcatctt catcaccccc accagcgacc
     1861 tgtattggtt caacctgagc ggcgccttta tcgccaagag cgtgggcgtg acattcgtgg
     1921 ataccaagtt cacctccagc cgggataccg ccaaccgggg cacatctgtg ctgatggccg
     1981 ccagcacctc cgccgtgtac accgtgacca cctccgacgc caccatcagc tacgccctgg
     2041 tgtccggcaa agaagtgaag tcctgcaacc ccgccctgaa caacgtggac agcgatacca
     2101 gccccacctt ctgcggaatc gcccggatct accccctgtc caccgacatg gtgtatatga
     2161 ccacaggcgg cgctagcgtc gtgcgggcca tcatcgtgtc caataccacc atctccgaca
     2221 ccatcacccg gacccccttc cctgtgtact tcctggacaa cagctccatc atgcccctga
     2281 tcctggatgg catgaactac gagctggtgt ctaactccaa catccccttt ccctacgtgg
     2341 ccatcaatca cagcaccccc aaggtggacg actccacctg ggacacaatc ttctccgtgg
     2401 atgtgtcaaa ccggttcttt agcacagtgt ccagagccgc cgtgatcggc acccctttca
     2461 tgagcagcct gcacggactg caggcctact acaaccggac caaccacatc ctgttcggcg
     2521 accccaacgt gctgcccatg tgcaacctga ccaagatgca gatgatcgag cgggccgtgg
     2581 ctgccgatgc cagagctgcc ctgcagtacc cctacatcta caccagcaag gcccagaact
     2641 tcaccgtgaa cgcccacccc aatctgacac tgctgaagct gctgatgccc taccctttcg
     2701 gcgagattct gaacgagagc ggcttcttcg agaacaccac cacccccgcc tccctggcca
     2761 atgtgcactt caacaccaca atgctggccg ctgtgcggaa cgcctacacc cccgatttcg
     2821 tgtacgactg catcttcgcc ggcaacgcct tccccttcca cattctgacc gccgctcagc
     2881 agcagctcgt gcggtggatc atctatacag ccatccagga acagctggcc aaatgcgccg
     2941 agaacagccc cagctacacc ggcagcggca gctcctccag cgacagccac gacgatatgg
     3001 tgcccggctg tgtgcccaga gtgggcatcg ataacctgac agagctggtc atgcccggca
     3061 tgccttacag caactacaac atcaccgtgt tcatccccga gtccctgcac tacaacttca
     3121 gcatctcccg gtgcctggac ggcaccgact ggaccaacgt gacagattac ctgcagaatg
     3181 ccaccacaac cagaacccgg aagtgtggca caggcgcgcc cggcagcctg caccacatcc
     3241 tggacgccca gaagatgctg tggaaccaca gggacaggaa cctgcctcct ctggcccctc
     3301 tgggccccca ccaccatcac caccattgag tctagacacg tgattaatta aggatccccc
     3361 gacctcgacc tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt
     3421 tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttggtcg agatccctcg
     3481 gagatctcta gctagaggat cgatccccgc cccggacgaa ctaaacctga ctacgacatc
     3541 tctgcccctt cttcgcgggg cagtgcatgt aatcccttca gttggttggt acaacttgcc
     3601 aactgggccc tgttccacat gtgacacggg gggggaccaa acacaaaggg gttctctgac
     3661 tgtagttgac atccttataa atggatgtgc acatttgcca acactgagtg gctttcatcc
     3721 tggagcagac tttgcagtct gtggactgca acacaacatt gcctttatgt gtaactcttg
     3781 gctgaagctc ttacaccaat gctgggggac atgtacctcc caggggccca ggaagactac
     3841 gggaggctac accaacgtca atcagagggg cctgtgtagc taccgataag cggaccctca
     3901 agagggcatt agcaatagtg tttataaggc ccccttgtta accctaaacg ggtagcatat
     3961 gcttcccggg tagtagtata tactatccag actaacccta attcaatagc atatgttacc
     4021 caacgggaag catatgctat cgaattaggg ttagtaaaag ggtcctaagg aacagcgata
     4081 tctcccaccc catgagctgt cacggtttta tttacatggg gtcaggattc cacgagggta
     4141 gtgaaccatt ttagtcacaa gggcagtggc tgaagatcaa ggagcgggca gtgaactctc
     4201 ctgaatcttc gcctgcttct tcattctcct tcgtttagct aatagaataa ctgctgagtt
     4261 gtgaacagta aggtgtatgt gaggtgctcg aaaacaaggt ttcaggtgac gcccccagaa
     4321 taaaatttgg acggggggtt cagtggtggc attgtgctat gacaccaata taaccctcac
     4381 aaaccccttg ggcaataaat actagtgtag gaatgaaaca ttctgaatat ctttaacaat
     4441 agaaatccat ggggtgggga caagccgtaa agactggatg tccatctcac acgaatttat
     4501 ggctatgggc aacacataat cctagtgcaa tatgatactg gggttattaa gatgtgtccc
     4561 aggcagggac caagacaggt gaaccatgtt gttacactct atttgtaaca aggggaaaga
     4621 gagtggacgc cgacagcagc ggactccact ggttgtctct aacacccccg aaaattaaac
     4681 ggggctccac gccaatgggg cccataaaca aagacaagtg gccactcttt tttttgaaat
     4741 tgtggagtgg gggcacgcgt cagcccccac acgccgccct gcggttttgg actgtaaaat
     4801 aagggtgtaa taacttggct gattgtaacc ccgctaacca ctgcggtcaa accacttgcc
     4861 cacaaaacca ctaatggcac cccggggaat acctgcataa gtaggtgggc gggccaagat
     4921 aggggcgcga ttgctgcgat ctggaggaca aattacacac acttgcgcct gagcgccaag
     4981 cacagggttg ttggtcctca tattcacgag gtcgctgaga gcacggtggg ctaatgttgc
     5041 catgggtagc atatactacc caaatatctg gatagcatat gctatcctaa tctatatctg
     5101 ggtagcatag gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg
     5161 ggtagtatat gctatcctaa tttatatctg ggtagcatag gctatcctaa tctatatctg
     5221 ggtagcatat gctatcctaa tctatatctg ggtagtatat gctatcctaa tctgtatccg
     5281 ggtagcatat gctatcctaa tagagattag ggtagtatat gctatcctaa tttatatctg
     5341 ggtagcatat actacccaaa tatctggata gcatatgcta tcctaatcta tatctgggta
     5401 gcatatgcta tcctaatcta tatctgggta gcataggcta tcctaatcta tatctgggta
     5461 gcatatgcta tcctaatcta tatctgggta gtatatgcta tcctaattta tatctgggta
     5521 gcataggcta tcctaatcta tatctgggta gcatatgcta tcctaatcta tatctgggta
     5581 gtatatgcta tcctaatctg tatccgggta gcatatgcta tcctcatgat aagctgtcaa
     5641 acatgagaat tttcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     5701 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     5761 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     5821 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     5881 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     5941 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     6001 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     6061 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag
     6121 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     6181 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     6241 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     6301 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     6361 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg
     6421 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     6481 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     6541 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     6601 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     6661 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     6721 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     6781 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     6841 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     6901 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     6961 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     7021 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     7081 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     7141 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     7201 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     7261 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     7321 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     7381 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     7441 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     7501 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     7561 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     7621 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     7681 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     7741 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     7801 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     7861 cacacaggaa acagctatga ccatgattac gccaagctct agctagaggt cgaccaattc
     7921 tcatgtttga cagcttatca tcgcagatcc gggcaacgtt gttgccattg ctgcaggcgc
     7981 agaactggta ggtatggaag atctatacat tgaatcaata ttggcaatta gccatattag
     8041 tcattggtta tatagcataa atcaatattg gctattggcc attgcatacg ttgtatctat
     8101 atcataatat gtacatttat attggctcat gtccaatatg accgc
//
