LOCUS       sequence-274890-        7479 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 7479)
  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 7479)
  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: D46_Ld_362700
FEATURES             Location/Qualifiers
     source          1..7479
                     /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             2637..2654
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2706..2761
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2760..2779)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4997..5015)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5083..5187
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5188..5256,5257..6048)
                     /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(5406..5425)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6219..6807
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6708..6727
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6961..6978
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7095..7116
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7131..7136,7137..7154,7155..7161)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7169..7185
                     /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     7174..7196
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7193..7209
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7193..7209
                     /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 ccatgtacaa ggtggaactg gtgcaggtcg tgcaccggca cggcgccaga tcccccctgg
     1261 tggacgacaa ccacaccctg atctgcggca ccgagttccc ttgcggcttc ctgaactacg
     1321 agggccaggc catgctggtc aagctgggcg agtacctgag acaccggtac accgaggacc
     1381 gcagcgtggt gtccaagccc tacttcccct acagctggta caacctgagc atcagctaca
     1441 cccggtccac cgacgtgctg cggaccctgc agtctgccaa cggcctgctg agaggcctgt
     1501 tccccaacaa cagcaacttc ttccccgcca tccacaccgt gggcttcgag gaagatgtgc
     1561 tgctgcacag catgatggtg cccatgatcc gggccagatt caactacgcc aaagaggaac
     1621 tgcgggccgt ctgcgaccag atgctggacc ggctgatgcc cttcgaccag ctgcaggccg
     1681 tggccgccga agtgcacagc cagggctttt gcgccaacta cacactgcgg agcagatgcg
     1741 ccgagcggct gtgcgacatc ggcagagcct acgagcccac cggcagactg gaacggctgc
     1801 ccctgctgtc cagacacctg gacgatgtgt gcgccgtgac cgccatgagc agccactttt
     1861 gcttcgccta caacgccagc aaccccgtgc accagaagca gggcgctccc ttctaccacc
     1921 tggcccggct gctggtgtct aacatgctgg cccaccagca gcgggaaacc gccccaccct
     1981 acaagctgta cgagtacagc gcccacgaca ccaccatcag ccccctggcc gtgtccttcg
     2041 gcgacagcag cctggaagct atgctgcccc ccttcggcac cgccttcatc atcgagctgc
     2101 tgagcctgac cgacgcccct gccgccagca gcagattcta cgtcagactg ctgcggggcc
     2161 acagcggcgt gcggcccgag agcaatttca ccttcgccct gagccacttc gacatgcggt
     2221 gcaaggacgc cacaggcaac acctatatcg ccaccgacaa catctgcccc ttcaccgact
     2281 tcgagcggtt catcaacagc acagccccta ccagccccat gggcacctgt tacctggacc
     2341 ccggactgct gttccggatg gactgccctg tggacgccgt gggcgacaac agaagcctga
     2401 gcaaggattg cctgttctac cggaagcact gcagcaatta cagctgtggc accggctact
     2461 acctggatgc catcgactac ggctgtcacc ggatccctgc caacaactcc accaccgtgt
     2521 ccagcccaat gtccagcgga ggcgcgcccg gcagcctgca ccacatcctg gacgcccaga
     2581 agatgctgtg gaaccacagg gacaggaacc tgcctcctct ggcccctctg ggcccccacc
     2641 accatcacca ccattgagtc tagacacgtg attaattaag gatcccccga cctcgacctc
     2701 tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc
     2761 actcggaagg acatatggga gggcaaatca tttggtcgag atccctcgga gatctctagc
     2821 tagaggatcg atccccgccc cggacgaact aaacctgact acgacatctc tgccccttct
     2881 tcgcggggca gtgcatgtaa tcccttcagt tggttggtac aacttgccaa ctgggccctg
     2941 ttccacatgt gacacggggg gggaccaaac acaaaggggt tctctgactg tagttgacat
     3001 ccttataaat ggatgtgcac atttgccaac actgagtggc tttcatcctg gagcagactt
     3061 tgcagtctgt ggactgcaac acaacattgc ctttatgtgt aactcttggc tgaagctctt
     3121 acaccaatgc tgggggacat gtacctccca ggggcccagg aagactacgg gaggctacac
     3181 caacgtcaat cagaggggcc tgtgtagcta ccgataagcg gaccctcaag agggcattag
     3241 caatagtgtt tataaggccc ccttgttaac cctaaacggg tagcatatgc ttcccgggta
     3301 gtagtatata ctatccagac taaccctaat tcaatagcat atgttaccca acgggaagca
     3361 tatgctatcg aattagggtt agtaaaaggg tcctaaggaa cagcgatatc tcccacccca
     3421 tgagctgtca cggttttatt tacatggggt caggattcca cgagggtagt gaaccatttt
     3481 agtcacaagg gcagtggctg aagatcaagg agcgggcagt gaactctcct gaatcttcgc
     3541 ctgcttcttc attctccttc gtttagctaa tagaataact gctgagttgt gaacagtaag
     3601 gtgtatgtga ggtgctcgaa aacaaggttt caggtgacgc ccccagaata aaatttggac
     3661 ggggggttca gtggtggcat tgtgctatga caccaatata accctcacaa accccttggg
     3721 caataaatac tagtgtagga atgaaacatt ctgaatatct ttaacaatag aaatccatgg
     3781 ggtggggaca agccgtaaag actggatgtc catctcacac gaatttatgg ctatgggcaa
     3841 cacataatcc tagtgcaata tgatactggg gttattaaga tgtgtcccag gcagggacca
     3901 agacaggtga accatgttgt tacactctat ttgtaacaag gggaaagaga gtggacgccg
     3961 acagcagcgg actccactgg ttgtctctaa cacccccgaa aattaaacgg ggctccacgc
     4021 caatggggcc cataaacaaa gacaagtggc cactcttttt tttgaaattg tggagtgggg
     4081 gcacgcgtca gcccccacac gccgccctgc ggttttggac tgtaaaataa gggtgtaata
     4141 acttggctga ttgtaacccc gctaaccact gcggtcaaac cacttgccca caaaaccact
     4201 aatggcaccc cggggaatac ctgcataagt aggtgggcgg gccaagatag gggcgcgatt
     4261 gctgcgatct ggaggacaaa ttacacacac ttgcgcctga gcgccaagca cagggttgtt
     4321 ggtcctcata ttcacgaggt cgctgagagc acggtgggct aatgttgcca tgggtagcat
     4381 atactaccca aatatctgga tagcatatgc tatcctaatc tatatctggg tagcataggc
     4441 tatcctaatc tatatctggg tagcatatgc tatcctaatc tatatctggg tagtatatgc
     4501 tatcctaatt tatatctggg tagcataggc tatcctaatc tatatctggg tagcatatgc
     4561 tatcctaatc tatatctggg tagtatatgc tatcctaatc tgtatccggg tagcatatgc
     4621 tatcctaata gagattaggg tagtatatgc tatcctaatt tatatctggg tagcatatac
     4681 tacccaaata tctggatagc atatgctatc ctaatctata tctgggtagc atatgctatc
     4741 ctaatctata tctgggtagc ataggctatc ctaatctata tctgggtagc atatgctatc
     4801 ctaatctata tctgggtagt atatgctatc ctaatttata tctgggtagc ataggctatc
     4861 ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt atatgctatc
     4921 ctaatctgta tccgggtagc atatgctatc ctcatgataa gctgtcaaac atgagaattt
     4981 tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     5041 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     5101 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     5161 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     5221 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     5281 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     5341 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     5401 gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc
     5461 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     5521 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     5581 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     5641 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     5701 ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta
     5761 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     5821 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     5881 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     5941 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     6001 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     6061 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     6121 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     6181 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     6241 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     6301 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     6361 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     6421 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     6481 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     6541 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     6601 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     6661 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     6721 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     6781 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     6841 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     6901 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     6961 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     7021 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     7081 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
     7141 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     7201 agctatgacc atgattacgc caagctctag ctagaggtcg accaattctc atgtttgaca
     7261 gcttatcatc gcagatccgg gcaacgttgt tgccattgct gcaggcgcag aactggtagg
     7321 tatggaagat ctatacattg aatcaatatt ggcaattagc catattagtc attggttata
     7381 tagcataaat caatattggc tattggccat tgcatacgtt gtatctatat cataatatgt
     7441 acatttatat tggctcatgt ccaatatgac cgccatgtt
//
