LOCUS       sequence-274471-        7071 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 7071)
  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 7071)
  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: D1_Ld_290860
FEATURES             Location/Qualifiers
     source          1..7071
                     /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             2229..2246
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2298..2353
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2352..2371)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4589..4607)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4675..4779
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4780..4848,4849..5640)
                     /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(4998..5017)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5811..6399
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6300..6319
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6553..6570
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6687..6708
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6723..6728,6729..6746,6747..6753)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6761..6777
                     /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     6766..6788
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6785..6801
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6785..6801
                     /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 gccaccaagc
     1201 ccagcgattt cccactgctg ggcaagagct tcgtggccga gatcaacagc aaggccagag
     1261 gccagtggac cgccagcgcc gacaatggct acctggtgtc cggcaagagc ctggaagagg
     1321 tccgaaagct gatgggcgtg accgacatga gcaccgaggc cgtgcccccc agaaacttca
     1381 gcgtggacga gatgcagcag gacctgcccg agttcttcga cgccgccgag cactggccta
     1441 tgtgcgtgac catcagcgag atccgggacc agagcaactg cggcagctgc tgggccattg
     1501 ccgccgtgga agccatcagc gaccggtact gtaccctggg cggcgtgccc gacagacgga
     1561 tcagcaccag caacctgctg agctgctgct tcatctgcgg cttcggctgc tacggcggca
     1621 tccccacaat ggcctggctg tggtgggtct gggtcggaat caccaccgaa gtgtgccagc
     1681 cctacccctt cggcccttgc agccaccacg gcaacagcga caagtacccc ccctgcccca
     1741 acaccatcta cgacaccccc aagtgcaaca ccacatgcga gaagtccgag atggacctgg
     1801 tcaagtacaa gggcggcacc agctacagcg tgaagggcga gaaagaactg atgatcgagc
     1861 tgatgaccaa cggccccctg gaagtgacca tgcaggtcta cagcgacttc gtgggctaca
     1921 agagcggcgt gtacaagcac gtgtccggcg acctgctggg cggccacgct gtgaaactcg
     1981 tgggctgggg aacccagggc ggagtgccct actggaagat cgccaacagc tggaacaccg
     2041 actggggcga caagggctac ttcctgatcc agcggggcag caacgagtgc ggcatcgagt
     2101 ctggcggcgt ggccggaaca ccagcccagg aaggcgcgcc cggcagcctg caccacatcc
     2161 tggacgccca gaagatgctg tggaaccaca gggacaggaa cctgcctcct ctggcccctc
     2221 tgggccccca ccaccatcac caccattgag tctagacacg tgattaatta aggatccccc
     2281 gacctcgacc tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt
     2341 tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttggtcg agatccctcg
     2401 gagatctcta gctagaggat cgatccccgc cccggacgaa ctaaacctga ctacgacatc
     2461 tctgcccctt cttcgcgggg cagtgcatgt aatcccttca gttggttggt acaacttgcc
     2521 aactgggccc tgttccacat gtgacacggg gggggaccaa acacaaaggg gttctctgac
     2581 tgtagttgac atccttataa atggatgtgc acatttgcca acactgagtg gctttcatcc
     2641 tggagcagac tttgcagtct gtggactgca acacaacatt gcctttatgt gtaactcttg
     2701 gctgaagctc ttacaccaat gctgggggac atgtacctcc caggggccca ggaagactac
     2761 gggaggctac accaacgtca atcagagggg cctgtgtagc taccgataag cggaccctca
     2821 agagggcatt agcaatagtg tttataaggc ccccttgtta accctaaacg ggtagcatat
     2881 gcttcccggg tagtagtata tactatccag actaacccta attcaatagc atatgttacc
     2941 caacgggaag catatgctat cgaattaggg ttagtaaaag ggtcctaagg aacagcgata
     3001 tctcccaccc catgagctgt cacggtttta tttacatggg gtcaggattc cacgagggta
     3061 gtgaaccatt ttagtcacaa gggcagtggc tgaagatcaa ggagcgggca gtgaactctc
     3121 ctgaatcttc gcctgcttct tcattctcct tcgtttagct aatagaataa ctgctgagtt
     3181 gtgaacagta aggtgtatgt gaggtgctcg aaaacaaggt ttcaggtgac gcccccagaa
     3241 taaaatttgg acggggggtt cagtggtggc attgtgctat gacaccaata taaccctcac
     3301 aaaccccttg ggcaataaat actagtgtag gaatgaaaca ttctgaatat ctttaacaat
     3361 agaaatccat ggggtgggga caagccgtaa agactggatg tccatctcac acgaatttat
     3421 ggctatgggc aacacataat cctagtgcaa tatgatactg gggttattaa gatgtgtccc
     3481 aggcagggac caagacaggt gaaccatgtt gttacactct atttgtaaca aggggaaaga
     3541 gagtggacgc cgacagcagc ggactccact ggttgtctct aacacccccg aaaattaaac
     3601 ggggctccac gccaatgggg cccataaaca aagacaagtg gccactcttt tttttgaaat
     3661 tgtggagtgg gggcacgcgt cagcccccac acgccgccct gcggttttgg actgtaaaat
     3721 aagggtgtaa taacttggct gattgtaacc ccgctaacca ctgcggtcaa accacttgcc
     3781 cacaaaacca ctaatggcac cccggggaat acctgcataa gtaggtgggc gggccaagat
     3841 aggggcgcga ttgctgcgat ctggaggaca aattacacac acttgcgcct gagcgccaag
     3901 cacagggttg ttggtcctca tattcacgag gtcgctgaga gcacggtggg ctaatgttgc
     3961 catgggtagc atatactacc caaatatctg gatagcatat gctatcctaa tctatatctg
     4021 ggtagcatag gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg
     4081 ggtagtatat gctatcctaa tttatatctg ggtagcatag gctatcctaa tctatatctg
     4141 ggtagcatat gctatcctaa tctatatctg ggtagtatat gctatcctaa tctgtatccg
     4201 ggtagcatat gctatcctaa tagagattag ggtagtatat gctatcctaa tttatatctg
     4261 ggtagcatat actacccaaa tatctggata gcatatgcta tcctaatcta tatctgggta
     4321 gcatatgcta tcctaatcta tatctgggta gcataggcta tcctaatcta tatctgggta
     4381 gcatatgcta tcctaatcta tatctgggta gtatatgcta tcctaattta tatctgggta
     4441 gcataggcta tcctaatcta tatctgggta gcatatgcta tcctaatcta tatctgggta
     4501 gtatatgcta tcctaatctg tatccgggta gcatatgcta tcctcatgat aagctgtcaa
     4561 acatgagaat tttcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     4621 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4681 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4741 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     4801 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     4861 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     4921 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     4981 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag
     5041 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     5101 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     5161 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     5221 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     5281 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg
     5341 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     5401 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     5461 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     5521 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     5581 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     5641 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     5701 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5761 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5821 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5881 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5941 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     6001 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     6061 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     6121 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     6181 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     6241 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6301 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6361 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6421 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     6481 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     6541 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     6601 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     6661 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     6721 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     6781 cacacaggaa acagctatga ccatgattac gccaagctct agctagaggt cgaccaattc
     6841 tcatgtttga cagcttatca tcgcagatcc gggcaacgtt gttgccattg ctgcaggcgc
     6901 agaactggta ggtatggaag atctatacat tgaatcaata ttggcaatta gccatattag
     6961 tcattggtta tatagcataa atcaatattg gctattggcc attgcatacg ttgtatctat
     7021 atcataatat gtacatttat attggctcat gtccaatatg accgccatgt t
//
