LOCUS       sequence-262380-        6573 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 6573)
  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 6573)
  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: D88_Ld_320450
FEATURES             Location/Qualifiers
     source          1..6573
                     /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             1737..1754
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1806..1861
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1860..1879)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4097..4115)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4183..4287
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4288..4356,4357..5148)
                     /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(4506..4525)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5319..5907
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5808..5827
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6061..6078
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6195..6216
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6231..6236,6237..6254,6255..6261)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6269..6285
                     /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     6274..6296
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6293..6309
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6293..6309
                     /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 ctacctggaa ctcaactcct gaaaagtatg gcgagtctgc agctgcgagc ggagccacag
     1261 tgttgccggg tgtggacctc gctcgcaggg ccgtctcccc aaatcaggaa tttccagcga
     1321 cctggtctga agacgaacct ctgtcaccat ccaaccctag agtttgctgg cgcgaacact
     1381 actcactgga tgctcaaaag ccgtattacc aaaatattaa gacaatggaa gtgacatggg
     1441 atattcccga aggctttgta accagatttc cacgacttta tgcaagcaac ggctaccatg
     1501 tcgacgcgac aggagcagtg tttcggtcct caaccgccag tgagccaccc gcaaatggaa
     1561 ccgtcgggac aacagtcgca aaagaggccg gtgtgaaagc agtcagtctg aaacagcgac
     1621 tggccgccta tggagctggc ggcgcgcccg gcagcctgca ccacatcctg gacgcccaga
     1681 agatgctgtg gaaccacagg gacaggaacc tgcctcctct ggcccctctg ggcccccacc
     1741 accatcacca ccattgagtc tagacacgtg attaattaag gatcccccga cctcgacctc
     1801 tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc
     1861 actcggaagg acatatggga gggcaaatca tttggtcgag atccctcgga gatctctagc
     1921 tagaggatcg atccccgccc cggacgaact aaacctgact acgacatctc tgccccttct
     1981 tcgcggggca gtgcatgtaa tcccttcagt tggttggtac aacttgccaa ctgggccctg
     2041 ttccacatgt gacacggggg gggaccaaac acaaaggggt tctctgactg tagttgacat
     2101 ccttataaat ggatgtgcac atttgccaac actgagtggc tttcatcctg gagcagactt
     2161 tgcagtctgt ggactgcaac acaacattgc ctttatgtgt aactcttggc tgaagctctt
     2221 acaccaatgc tgggggacat gtacctccca ggggcccagg aagactacgg gaggctacac
     2281 caacgtcaat cagaggggcc tgtgtagcta ccgataagcg gaccctcaag agggcattag
     2341 caatagtgtt tataaggccc ccttgttaac cctaaacggg tagcatatgc ttcccgggta
     2401 gtagtatata ctatccagac taaccctaat tcaatagcat atgttaccca acgggaagca
     2461 tatgctatcg aattagggtt agtaaaaggg tcctaaggaa cagcgatatc tcccacccca
     2521 tgagctgtca cggttttatt tacatggggt caggattcca cgagggtagt gaaccatttt
     2581 agtcacaagg gcagtggctg aagatcaagg agcgggcagt gaactctcct gaatcttcgc
     2641 ctgcttcttc attctccttc gtttagctaa tagaataact gctgagttgt gaacagtaag
     2701 gtgtatgtga ggtgctcgaa aacaaggttt caggtgacgc ccccagaata aaatttggac
     2761 ggggggttca gtggtggcat tgtgctatga caccaatata accctcacaa accccttggg
     2821 caataaatac tagtgtagga atgaaacatt ctgaatatct ttaacaatag aaatccatgg
     2881 ggtggggaca agccgtaaag actggatgtc catctcacac gaatttatgg ctatgggcaa
     2941 cacataatcc tagtgcaata tgatactggg gttattaaga tgtgtcccag gcagggacca
     3001 agacaggtga accatgttgt tacactctat ttgtaacaag gggaaagaga gtggacgccg
     3061 acagcagcgg actccactgg ttgtctctaa cacccccgaa aattaaacgg ggctccacgc
     3121 caatggggcc cataaacaaa gacaagtggc cactcttttt tttgaaattg tggagtgggg
     3181 gcacgcgtca gcccccacac gccgccctgc ggttttggac tgtaaaataa gggtgtaata
     3241 acttggctga ttgtaacccc gctaaccact gcggtcaaac cacttgccca caaaaccact
     3301 aatggcaccc cggggaatac ctgcataagt aggtgggcgg gccaagatag gggcgcgatt
     3361 gctgcgatct ggaggacaaa ttacacacac ttgcgcctga gcgccaagca cagggttgtt
     3421 ggtcctcata ttcacgaggt cgctgagagc acggtgggct aatgttgcca tgggtagcat
     3481 atactaccca aatatctgga tagcatatgc tatcctaatc tatatctggg tagcataggc
     3541 tatcctaatc tatatctggg tagcatatgc tatcctaatc tatatctggg tagtatatgc
     3601 tatcctaatt tatatctggg tagcataggc tatcctaatc tatatctggg tagcatatgc
     3661 tatcctaatc tatatctggg tagtatatgc tatcctaatc tgtatccggg tagcatatgc
     3721 tatcctaata gagattaggg tagtatatgc tatcctaatt tatatctggg tagcatatac
     3781 tacccaaata tctggatagc atatgctatc ctaatctata tctgggtagc atatgctatc
     3841 ctaatctata tctgggtagc ataggctatc ctaatctata tctgggtagc atatgctatc
     3901 ctaatctata tctgggtagt atatgctatc ctaatttata tctgggtagc ataggctatc
     3961 ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt atatgctatc
     4021 ctaatctgta tccgggtagc atatgctatc ctcatgataa gctgtcaaac atgagaattt
     4081 tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     4141 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     4201 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     4261 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     4321 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     4381 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     4441 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     4501 gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc
     4561 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     4621 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     4681 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     4741 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     4801 ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta
     4861 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     4921 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     4981 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     5041 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     5101 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     5161 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     5221 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5281 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     5341 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     5401 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     5461 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     5521 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     5581 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     5641 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     5701 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     5761 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     5821 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     5881 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     5941 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     6001 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     6061 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     6121 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     6181 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
     6241 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     6301 agctatgacc atgattacgc caagctctag ctagaggtcg accaattctc atgtttgaca
     6361 gcttatcatc gcagatccgg gcaacgttgt tgccattgct gcaggcgcag aactggtagg
     6421 tatggaagat ctatacattg aatcaatatt ggcaattagc catattagtc attggttata
     6481 tagcataaat caatattggc tattggccat tgcatacgtt gtatctatat cataatatgt
     6541 acatttatat tggctcatgt ccaatatgac cgc
//
