LOCUS       sequence-274886-        7266 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 7266)
  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 7266)
  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: D59_Ld_312380
FEATURES             Location/Qualifiers
     source          1..7266
                     /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"
     regulatory      808..817
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2044..2061
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2113..2168
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2167..2186)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4404..4422)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4490..4594
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4595..4663,4664..5455)
                     /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(4813..4832)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5626..6214
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6115..6134
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6368..6385
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6502..6523
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6538..6543,6544..6561,6562..6568)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6576..6592
                     /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     6581..6603
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6600..6616
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6600..6616
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        6887..7266
                     /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 gccatggcca gagccagatt tctgcagctg
      841 ctgctggtca ccctgaccct gctgtctaca gccgccctgc ctgtgtccgc ctggtggtcc
      901 aagggccaca tgagcgtggc cctgatcgcc aagagacaca tgggcgccag cctggtggaa
      961 aaggccgagc tggccgccaa ggtgctgagc ttcagcggcc cctaccccaa gagccccgac
     1021 atggtgcaga ccgccccctg ggccgacgac atcaagacca tcggcctgaa aaccctgagc
     1081 acctggcact acatcaccac cccctactac accgacgagg acttcaccct ggacgtgtcc
     1141 cccgtgcaga ccgtgaacgt ggccagcgtg atccccatgc tgcagacagc catcgagaag
     1201 cccaccgcca acagcgacgt gatcgtgcag agcctggccc tgctgctgca cttcatgggc
     1261 gacatccacc agcccctgca caacgtgaac ctgttcagca accagtaccc cgagagcgac
     1321 ctgggcggca acaagcagct ggtggtcatc gacagcaagg gcaccaagat gctgctgcat
     1381 gcctactggg acagcatggc cgagggcaag agcggcgagg acgtgcccag acctctgagc
     1441 gaggccgact acgacgacct gaacaacttc gccgactacc tggaagccac ctacgccagc
     1501 accctgaccg acaaagaaaa gaacctggtg gacaccaccg agatcagcaa agagacattc
     1561 gacctggccc tgaagtacgc ctaccctggc gccgacaatg gcgccaccct gagcgacgag
     1621 tacaagacca acgccaagaa gatcagcgag agacaggtgc tgctggccgg ctaccggctg
     1681 gccaaaatgc tgaacaccac cctgaagtcc gtgtccatgg ataccatcct gcagggcctg
     1741 aagtccatcc agagcgaggt ggacaccgag aacaaggccg aggtgcacaa ccactacgac
     1801 cagaagggca tctctgccgc cgtgaccgcc attgtggccg tggccctgtt tatcgccggc
     1861 atcatcattg ccaccctggt ggtgctggct ctgaagtgct acctgcccaa gcgggacaga
     1921 ttcggcagct acgagcacgt ggccctgggc gcgcccggca gcctgcacca catcctggac
     1981 gcccagaaga tgctgtggaa ccacagggac aggaacctgc ctcctctggc ccctctgggc
     2041 ccccaccacc atcaccacca ttgagtctag acacgtgatt aattaaggat cccccgacct
     2101 cgacctctgg ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt
     2161 gtctctcact cggaaggaca tatgggaggg caaatcattt ggtcgagatc cctcggagat
     2221 ctctagctag aggatcgatc cccgccccgg acgaactaaa cctgactacg acatctctgc
     2281 cccttcttcg cggggcagtg catgtaatcc cttcagttgg ttggtacaac ttgccaactg
     2341 ggccctgttc cacatgtgac acgggggggg accaaacaca aaggggttct ctgactgtag
     2401 ttgacatcct tataaatgga tgtgcacatt tgccaacact gagtggcttt catcctggag
     2461 cagactttgc agtctgtgga ctgcaacaca acattgcctt tatgtgtaac tcttggctga
     2521 agctcttaca ccaatgctgg gggacatgta cctcccaggg gcccaggaag actacgggag
     2581 gctacaccaa cgtcaatcag aggggcctgt gtagctaccg ataagcggac cctcaagagg
     2641 gcattagcaa tagtgtttat aaggccccct tgttaaccct aaacgggtag catatgcttc
     2701 ccgggtagta gtatatacta tccagactaa ccctaattca atagcatatg ttacccaacg
     2761 ggaagcatat gctatcgaat tagggttagt aaaagggtcc taaggaacag cgatatctcc
     2821 caccccatga gctgtcacgg ttttatttac atggggtcag gattccacga gggtagtgaa
     2881 ccattttagt cacaagggca gtggctgaag atcaaggagc gggcagtgaa ctctcctgaa
     2941 tcttcgcctg cttcttcatt ctccttcgtt tagctaatag aataactgct gagttgtgaa
     3001 cagtaaggtg tatgtgaggt gctcgaaaac aaggtttcag gtgacgcccc cagaataaaa
     3061 tttggacggg gggttcagtg gtggcattgt gctatgacac caatataacc ctcacaaacc
     3121 ccttgggcaa taaatactag tgtaggaatg aaacattctg aatatcttta acaatagaaa
     3181 tccatggggt ggggacaagc cgtaaagact ggatgtccat ctcacacgaa tttatggcta
     3241 tgggcaacac ataatcctag tgcaatatga tactggggtt attaagatgt gtcccaggca
     3301 gggaccaaga caggtgaacc atgttgttac actctatttg taacaagggg aaagagagtg
     3361 gacgccgaca gcagcggact ccactggttg tctctaacac ccccgaaaat taaacggggc
     3421 tccacgccaa tggggcccat aaacaaagac aagtggccac tctttttttt gaaattgtgg
     3481 agtgggggca cgcgtcagcc cccacacgcc gccctgcggt tttggactgt aaaataaggg
     3541 tgtaataact tggctgattg taaccccgct aaccactgcg gtcaaaccac ttgcccacaa
     3601 aaccactaat ggcaccccgg ggaatacctg cataagtagg tgggcgggcc aagatagggg
     3661 cgcgattgct gcgatctgga ggacaaatta cacacacttg cgcctgagcg ccaagcacag
     3721 ggttgttggt cctcatattc acgaggtcgc tgagagcacg gtgggctaat gttgccatgg
     3781 gtagcatata ctacccaaat atctggatag catatgctat cctaatctat atctgggtag
     3841 cataggctat cctaatctat atctgggtag catatgctat cctaatctat atctgggtag
     3901 tatatgctat cctaatttat atctgggtag cataggctat cctaatctat atctgggtag
     3961 catatgctat cctaatctat atctgggtag tatatgctat cctaatctgt atccgggtag
     4021 catatgctat cctaatagag attagggtag tatatgctat cctaatttat atctgggtag
     4081 catatactac ccaaatatct ggatagcata tgctatccta atctatatct gggtagcata
     4141 tgctatccta atctatatct gggtagcata ggctatccta atctatatct gggtagcata
     4201 tgctatccta atctatatct gggtagtata tgctatccta atttatatct gggtagcata
     4261 ggctatccta atctatatct gggtagcata tgctatccta atctatatct gggtagtata
     4321 tgctatccta atctgtatcc gggtagcata tgctatcctc atgataagct gtcaaacatg
     4381 agaattttct tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     4441 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     4501 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     4561 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     4621 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     4681 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     4741 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     4801 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact
     4861 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     4921 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     4981 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     5041 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     5101 agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg
     5161 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     5221 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     5281 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     5341 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     5401 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     5461 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     5521 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     5581 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     5641 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5701 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5761 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5821 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5881 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5941 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     6001 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     6061 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6121 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     6181 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     6241 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     6301 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     6361 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     6421 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     6481 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     6541 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     6601 aggaaacagc tatgaccatg attacgccaa gctctagcta gaggtcgacc aattctcatg
     6661 tttgacagct tatcatcgca gatccgggca acgttgttgc cattgctgca ggcgcagaac
     6721 tggtaggtat ggaagatcta tacattgaat caatattggc aattagccat attagtcatt
     6781 ggttatatag cataaatcaa tattggctat tggccattgc atacgttgta tctatatcat
     6841 aatatgtaca tttatattgg ctcatgtcca atatgaccgc catgttgaca ttgattattg
     6901 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
     6961 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
     7021 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
     7081 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
     7141 ccaagtccgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
     7201 tacatgacct tacgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
     7261 accatg
//
