LOCUS       sequence-262345-        7002 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 7002)
  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 7002)
  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: D53_Ld_365900
FEATURES             Location/Qualifiers
     source          1..7002
                     /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             2166..2183
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2235..2290
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2289..2308)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4526..4544)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4612..4716
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4717..4785,4786..5577)
                     /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(4935..4954)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5748..6336
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6237..6256
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6490..6507
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6624..6645
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6660..6665,6666..6683,6684..6690)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6698..6714
                     /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     6703..6725
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6722..6738
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6722..6738
                     /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 gaaagcagct gcccctgaac accccctacc cccccgtggt gcaggacgcc ctgagatgcg
     1261 acgtgtgcag cttcatcgtg gtcaacgccc tgtaccaggt ggaagccaag agagaggaac
     1321 agcagcggaa gcggctgcag gtccgagagg acgacgtgct ggaagaggtc gagaatatgt
     1381 gcgtgccctt caaggaccag ggccagtgga tcagacaggt ggccctgcag atggaagagg
     1441 tggcccccag caagagaggc gccgatagac ctgccgccgc tacccccaga taccacatga
     1501 gcgtgggcgt ggtggactac tacagcaagt gcggccggat ctgcgagaca gtggccgtgc
     1561 tgtgcgagga atggatggac agcggctaca tggacggctt cagcagccgg ctggtcaaag
     1621 aggccaaggc cggcagaaac atcagcgacg ccgcccacag ggacgtggtg ttcggcagct
     1681 tctgcgcccc cagcccccac tgcaagaaac acgccacctt cgtgcggaag ctggacatgg
     1741 ccctgaccaa ggacaccgag ctgagagaga gcatcgacgc cgaccggccc caggaaatcg
     1801 agacagagga acgcgagatg gaaaccatgc tgcaccggct gaccagagag cagggccaga
     1861 gcgccgacgt gttcagccgg gacgagatcc ggcggatgaa ggaagccttc gtcaccggca
     1921 acaaagagga cctgcaggcc gtcgacccca ccgccttcga cctgaccgac gacgagttca
     1981 gcaccctgca ggactacatg cggggcgagg aacggaagca gcagcagcgg cagagaaggc
     2041 ccagcgcccc tagcgacgcc gacgatctgg gcgcgcccgg cagcctgcac cacatcctgg
     2101 acgcccagaa gatgctgtgg aaccacaggg acaggaacct gcctcctctg gcccctctgg
     2161 gcccccacca ccatcaccac cattgagtct agacacgtga ttaattaagg atcccccgac
     2221 ctcgacctct ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt
     2281 gtgtctctca ctcggaagga catatgggag ggcaaatcat ttggtcgaga tccctcggag
     2341 atctctagct agaggatcga tccccgcccc ggacgaacta aacctgacta cgacatctct
     2401 gccccttctt cgcggggcag tgcatgtaat cccttcagtt ggttggtaca acttgccaac
     2461 tgggccctgt tccacatgtg acacgggggg ggaccaaaca caaaggggtt ctctgactgt
     2521 agttgacatc cttataaatg gatgtgcaca tttgccaaca ctgagtggct ttcatcctgg
     2581 agcagacttt gcagtctgtg gactgcaaca caacattgcc tttatgtgta actcttggct
     2641 gaagctctta caccaatgct gggggacatg tacctcccag gggcccagga agactacggg
     2701 aggctacacc aacgtcaatc agaggggcct gtgtagctac cgataagcgg accctcaaga
     2761 gggcattagc aatagtgttt ataaggcccc cttgttaacc ctaaacgggt agcatatgct
     2821 tcccgggtag tagtatatac tatccagact aaccctaatt caatagcata tgttacccaa
     2881 cgggaagcat atgctatcga attagggtta gtaaaagggt cctaaggaac agcgatatct
     2941 cccaccccat gagctgtcac ggttttattt acatggggtc aggattccac gagggtagtg
     3001 aaccatttta gtcacaaggg cagtggctga agatcaagga gcgggcagtg aactctcctg
     3061 aatcttcgcc tgcttcttca ttctccttcg tttagctaat agaataactg ctgagttgtg
     3121 aacagtaagg tgtatgtgag gtgctcgaaa acaaggtttc aggtgacgcc cccagaataa
     3181 aatttggacg gggggttcag tggtggcatt gtgctatgac accaatataa ccctcacaaa
     3241 ccccttgggc aataaatact agtgtaggaa tgaaacattc tgaatatctt taacaataga
     3301 aatccatggg gtggggacaa gccgtaaaga ctggatgtcc atctcacacg aatttatggc
     3361 tatgggcaac acataatcct agtgcaatat gatactgggg ttattaagat gtgtcccagg
     3421 cagggaccaa gacaggtgaa ccatgttgtt acactctatt tgtaacaagg ggaaagagag
     3481 tggacgccga cagcagcgga ctccactggt tgtctctaac acccccgaaa attaaacggg
     3541 gctccacgcc aatggggccc ataaacaaag acaagtggcc actctttttt ttgaaattgt
     3601 ggagtggggg cacgcgtcag cccccacacg ccgccctgcg gttttggact gtaaaataag
     3661 ggtgtaataa cttggctgat tgtaaccccg ctaaccactg cggtcaaacc acttgcccac
     3721 aaaaccacta atggcacccc ggggaatacc tgcataagta ggtgggcggg ccaagatagg
     3781 ggcgcgattg ctgcgatctg gaggacaaat tacacacact tgcgcctgag cgccaagcac
     3841 agggttgttg gtcctcatat tcacgaggtc gctgagagca cggtgggcta atgttgccat
     3901 gggtagcata tactacccaa atatctggat agcatatgct atcctaatct atatctgggt
     3961 agcataggct atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt
     4021 agtatatgct atcctaattt atatctgggt agcataggct atcctaatct atatctgggt
     4081 agcatatgct atcctaatct atatctgggt agtatatgct atcctaatct gtatccgggt
     4141 agcatatgct atcctaatag agattagggt agtatatgct atcctaattt atatctgggt
     4201 agcatatact acccaaatat ctggatagca tatgctatcc taatctatat ctgggtagca
     4261 tatgctatcc taatctatat ctgggtagca taggctatcc taatctatat ctgggtagca
     4321 tatgctatcc taatctatat ctgggtagta tatgctatcc taatttatat ctgggtagca
     4381 taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagta
     4441 tatgctatcc taatctgtat ccgggtagca tatgctatcc tcatgataag ctgtcaaaca
     4501 tgagaatttt cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     4561 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     4621 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4681 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     4741 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     4801 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     4861 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     4921 acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa
     4981 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     5041 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     5101 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     5161 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     5221 gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg
     5281 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     5341 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     5401 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     5461 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     5521 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     5581 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     5641 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5701 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     5761 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     5821 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     5881 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     5941 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     6001 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     6061 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     6121 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     6181 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     6241 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     6301 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     6361 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     6421 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     6481 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     6541 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     6601 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     6661 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     6721 acaggaaaca gctatgacca tgattacgcc aagctctagc tagaggtcga ccaattctca
     6781 tgtttgacag cttatcatcg cagatccggg caacgttgtt gccattgctg caggcgcaga
     6841 actggtaggt atggaagatc tatacattga atcaatattg gcaattagcc atattagtca
     6901 ttggttatat agcataaatc aatattggct attggccatt gcatacgttg tatctatatc
     6961 ataatatgta catttatatt ggctcatgtc caatatgacc gc
//
