LOCUS       sequence-262387-        6582 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 6582)
  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 6582)
  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: D95_Ld_290220
FEATURES             Location/Qualifiers
     source          1..6582
                     /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             1746..1763
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1815..1870
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1869..1888)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4106..4124)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4192..4296
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4297..4365,4366..5157)
                     /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(4515..4534)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5328..5916
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5817..5836
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6070..6087
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6204..6225
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6240..6245,6246..6263,6264..6270)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6278..6294
                     /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     6283..6305
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6302..6318
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6302..6318
                     /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 gcggccgcct
     1201 gggactcttc tgaagacgct aaagccatgg ccaaaagggc caagcatgag cagattcagt
     1261 tttgggaacg tgaggttaat attttgcggc agggagaact gactcgtgct tacaacaagc
     1321 tgtatcaagc tgaggccgct ctcgagagtg ccagggctaa acaaggcttc ttctacactc
     1381 gcccccagga taaagctacg atcagactgc tcgatgaaga ttaccgcagg actttggtcg
     1441 aggtgaaagc tcttaaggaa caagagcgac ttatcatggc caaactgaag cccctctacg
     1501 gggtggtgag tctgcatttc gcccaggaac agaagcgcac gatttctgag tctatcaaaa
     1561 ctgtgcagtc tcttagctat gacaatgctt ggtacagcag cctcttttct cttggggaag
     1621 ccgagagctt cagcgatatc attatgggtg gcgcgcccgg cagcctgcac cacatcctgg
     1681 acgcccagaa gatgctgtgg aaccacaggg acaggaacct gcctcctctg gcccctctgg
     1741 gcccccacca ccatcaccac cattgagtct agacacgtga ttaattaagg atcccccgac
     1801 ctcgacctct ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt
     1861 gtgtctctca ctcggaagga catatgggag ggcaaatcat ttggtcgaga tccctcggag
     1921 atctctagct agaggatcga tccccgcccc ggacgaacta aacctgacta cgacatctct
     1981 gccccttctt cgcggggcag tgcatgtaat cccttcagtt ggttggtaca acttgccaac
     2041 tgggccctgt tccacatgtg acacgggggg ggaccaaaca caaaggggtt ctctgactgt
     2101 agttgacatc cttataaatg gatgtgcaca tttgccaaca ctgagtggct ttcatcctgg
     2161 agcagacttt gcagtctgtg gactgcaaca caacattgcc tttatgtgta actcttggct
     2221 gaagctctta caccaatgct gggggacatg tacctcccag gggcccagga agactacggg
     2281 aggctacacc aacgtcaatc agaggggcct gtgtagctac cgataagcgg accctcaaga
     2341 gggcattagc aatagtgttt ataaggcccc cttgttaacc ctaaacgggt agcatatgct
     2401 tcccgggtag tagtatatac tatccagact aaccctaatt caatagcata tgttacccaa
     2461 cgggaagcat atgctatcga attagggtta gtaaaagggt cctaaggaac agcgatatct
     2521 cccaccccat gagctgtcac ggttttattt acatggggtc aggattccac gagggtagtg
     2581 aaccatttta gtcacaaggg cagtggctga agatcaagga gcgggcagtg aactctcctg
     2641 aatcttcgcc tgcttcttca ttctccttcg tttagctaat agaataactg ctgagttgtg
     2701 aacagtaagg tgtatgtgag gtgctcgaaa acaaggtttc aggtgacgcc cccagaataa
     2761 aatttggacg gggggttcag tggtggcatt gtgctatgac accaatataa ccctcacaaa
     2821 ccccttgggc aataaatact agtgtaggaa tgaaacattc tgaatatctt taacaataga
     2881 aatccatggg gtggggacaa gccgtaaaga ctggatgtcc atctcacacg aatttatggc
     2941 tatgggcaac acataatcct agtgcaatat gatactgggg ttattaagat gtgtcccagg
     3001 cagggaccaa gacaggtgaa ccatgttgtt acactctatt tgtaacaagg ggaaagagag
     3061 tggacgccga cagcagcgga ctccactggt tgtctctaac acccccgaaa attaaacggg
     3121 gctccacgcc aatggggccc ataaacaaag acaagtggcc actctttttt ttgaaattgt
     3181 ggagtggggg cacgcgtcag cccccacacg ccgccctgcg gttttggact gtaaaataag
     3241 ggtgtaataa cttggctgat tgtaaccccg ctaaccactg cggtcaaacc acttgcccac
     3301 aaaaccacta atggcacccc ggggaatacc tgcataagta ggtgggcggg ccaagatagg
     3361 ggcgcgattg ctgcgatctg gaggacaaat tacacacact tgcgcctgag cgccaagcac
     3421 agggttgttg gtcctcatat tcacgaggtc gctgagagca cggtgggcta atgttgccat
     3481 gggtagcata tactacccaa atatctggat agcatatgct atcctaatct atatctgggt
     3541 agcataggct atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt
     3601 agtatatgct atcctaattt atatctgggt agcataggct atcctaatct atatctgggt
     3661 agcatatgct atcctaatct atatctgggt agtatatgct atcctaatct gtatccgggt
     3721 agcatatgct atcctaatag agattagggt agtatatgct atcctaattt atatctgggt
     3781 agcatatact acccaaatat ctggatagca tatgctatcc taatctatat ctgggtagca
     3841 tatgctatcc taatctatat ctgggtagca taggctatcc taatctatat ctgggtagca
     3901 tatgctatcc taatctatat ctgggtagta tatgctatcc taatttatat ctgggtagca
     3961 taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagta
     4021 tatgctatcc taatctgtat ccgggtagca tatgctatcc tcatgataag ctgtcaaaca
     4081 tgagaatttt cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     4141 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     4201 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4261 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     4321 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     4381 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     4441 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     4501 acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa
     4561 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     4621 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     4681 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     4741 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     4801 gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg
     4861 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     4921 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     4981 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     5041 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     5101 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     5161 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     5221 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5281 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     5341 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     5401 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     5461 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     5521 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     5581 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     5641 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     5701 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     5761 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     5821 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     5881 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     5941 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     6001 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     6061 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     6121 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     6181 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     6241 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     6301 acaggaaaca gctatgacca tgattacgcc aagctctagc tagaggtcga ccaattctca
     6361 tgtttgacag cttatcatcg cagatccggg caacgttgtt gccattgctg caggcgcaga
     6421 actggtaggt atggaagatc tatacattga atcaatattg gcaattagcc atattagtca
     6481 ttggttatat agcataaatc aatattggct attggccatt gcatacgttg tatctatatc
     6541 ataatatgta catttatatt ggctcatgtc caatatgacc gc
//
