LOCUS       sequence-274901-        6978 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 6978)
  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 6978)
  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: D29_Ld_251870
FEATURES             Location/Qualifiers
     source          1..6978
                     /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             2136..2153
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2205..2260
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2259..2278)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4496..4514)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4582..4686
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4687..4755,4756..5547)
                     /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(4905..4924)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5718..6306
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6207..6226
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6460..6477
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6594..6615
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6630..6635,6636..6653,6654..6660)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6668..6684
                     /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     6673..6695
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6692..6708
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6692..6708
                     /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 gccaccgccc
     1201 tgtactacta cagcgtgtac gactacgaga agcacatgag catggtgcag cggaagtaca
     1261 gcggctacga ccccctgacc gactgcgcca ccccttttgg ccagctgctg ggcgtggccg
     1321 acgatgtgcc cgcctacagc aactgcaaca ccaagttcag cagcacctac atcaactacg
     1381 tgtccctgat ggaccccatg gacaacggca gacggggcga ccccagcgag acacggatcg
     1441 tgatgaccgc ctaccggtac accgccttcg actactgcat gcggtggctc gtgtggaacc
     1501 ggggcgtgat gccccggctg gtggaaaaca ccaaccagct ctggaaaacc gtggactact
     1561 tcaaccccgc caagcccgag caggggtggt ccgccgagta catcaccaac tacgaagagg
     1621 tcacaggcat ggaagagaga aagttcaacg cccccagacg cgccgacgcc atcgtgtacc
     1681 ggatggacaa gaacaccatc cctgccggcc acattgccgt ggtggtcaag gtggaagatg
     1741 acgtggaagc cgctggcggc cctgagaagc tgaaagaact gaagaagatg cggctgcacc
     1801 ccaggcgggt gtacgtggca gagcagaatt ggaagaacca gccctggggc ggacacaact
     1861 acagccgggt gctgcagttc aagtggcgcg ctgtggccga gaaggcctac gagggctgct
     1921 acgtggaccc cgacggcctg ggcatcatcg gcgtcgtcag agtgggcaag gccatgcccc
     1981 tgagagccgc cccagacccc taccaggaag ccctggacat ggataacgac ggcgacctgg
     2041 gcgcgcccgg cagcctgcac cacatcctgg acgcccagaa gatgctgtgg aaccacaggg
     2101 acaggaacct gcctcctctg gcccctctgg gcccccacca ccatcaccac cattgagtct
     2161 agacacgtga ttaattaagg atcccccgac ctcgacctct ggctaataaa ggaaatttat
     2221 tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag
     2281 ggcaaatcat ttggtcgaga tccctcggag atctctagct agaggatcga tccccgcccc
     2341 ggacgaacta aacctgacta cgacatctct gccccttctt cgcggggcag tgcatgtaat
     2401 cccttcagtt ggttggtaca acttgccaac tgggccctgt tccacatgtg acacgggggg
     2461 ggaccaaaca caaaggggtt ctctgactgt agttgacatc cttataaatg gatgtgcaca
     2521 tttgccaaca ctgagtggct ttcatcctgg agcagacttt gcagtctgtg gactgcaaca
     2581 caacattgcc tttatgtgta actcttggct gaagctctta caccaatgct gggggacatg
     2641 tacctcccag gggcccagga agactacggg aggctacacc aacgtcaatc agaggggcct
     2701 gtgtagctac cgataagcgg accctcaaga gggcattagc aatagtgttt ataaggcccc
     2761 cttgttaacc ctaaacgggt agcatatgct tcccgggtag tagtatatac tatccagact
     2821 aaccctaatt caatagcata tgttacccaa cgggaagcat atgctatcga attagggtta
     2881 gtaaaagggt cctaaggaac agcgatatct cccaccccat gagctgtcac ggttttattt
     2941 acatggggtc aggattccac gagggtagtg aaccatttta gtcacaaggg cagtggctga
     3001 agatcaagga gcgggcagtg aactctcctg aatcttcgcc tgcttcttca ttctccttcg
     3061 tttagctaat agaataactg ctgagttgtg aacagtaagg tgtatgtgag gtgctcgaaa
     3121 acaaggtttc aggtgacgcc cccagaataa aatttggacg gggggttcag tggtggcatt
     3181 gtgctatgac accaatataa ccctcacaaa ccccttgggc aataaatact agtgtaggaa
     3241 tgaaacattc tgaatatctt taacaataga aatccatggg gtggggacaa gccgtaaaga
     3301 ctggatgtcc atctcacacg aatttatggc tatgggcaac acataatcct agtgcaatat
     3361 gatactgggg ttattaagat gtgtcccagg cagggaccaa gacaggtgaa ccatgttgtt
     3421 acactctatt tgtaacaagg ggaaagagag tggacgccga cagcagcgga ctccactggt
     3481 tgtctctaac acccccgaaa attaaacggg gctccacgcc aatggggccc ataaacaaag
     3541 acaagtggcc actctttttt ttgaaattgt ggagtggggg cacgcgtcag cccccacacg
     3601 ccgccctgcg gttttggact gtaaaataag ggtgtaataa cttggctgat tgtaaccccg
     3661 ctaaccactg cggtcaaacc acttgcccac aaaaccacta atggcacccc ggggaatacc
     3721 tgcataagta ggtgggcggg ccaagatagg ggcgcgattg ctgcgatctg gaggacaaat
     3781 tacacacact tgcgcctgag cgccaagcac agggttgttg gtcctcatat tcacgaggtc
     3841 gctgagagca cggtgggcta atgttgccat gggtagcata tactacccaa atatctggat
     3901 agcatatgct atcctaatct atatctgggt agcataggct atcctaatct atatctgggt
     3961 agcatatgct atcctaatct atatctgggt agtatatgct atcctaattt atatctgggt
     4021 agcataggct atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt
     4081 agtatatgct atcctaatct gtatccgggt agcatatgct atcctaatag agattagggt
     4141 agtatatgct atcctaattt atatctgggt agcatatact acccaaatat ctggatagca
     4201 tatgctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagca
     4261 taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagta
     4321 tatgctatcc taatttatat ctgggtagca taggctatcc taatctatat ctgggtagca
     4381 tatgctatcc taatctatat ctgggtagta tatgctatcc taatctgtat ccgggtagca
     4441 tatgctatcc tcatgataag ctgtcaaaca tgagaatttt cttgaagacg aaagggcctc
     4501 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     4561 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     4621 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     4681 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     4741 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     4801 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     4861 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     4921 ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     4981 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     5041 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     5101 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     5161 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     5221 acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     5281 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     5341 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     5401 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     5461 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     5521 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     5581 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     5641 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     5701 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     5761 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     5821 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     5881 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     5941 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     6001 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     6061 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     6121 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     6181 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     6241 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     6301 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     6361 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     6421 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     6481 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     6541 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     6601 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     6661 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     6721 aagctctagc tagaggtcga ccaattctca tgtttgacag cttatcatcg cagatccggg
     6781 caacgttgtt gccattgctg caggcgcaga actggtaggt atggaagatc tatacattga
     6841 atcaatattg gcaattagcc atattagtca ttggttatat agcataaatc aatattggct
     6901 attggccatt gcatacgttg tatctatatc ataatatgta catttatatt ggctcatgtc
     6961 caatatgacc gccatgtt
//
