LOCUS       sequence-274903-        7299 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 7299)
  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 7299)
  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: D28_Ld_241170
FEATURES             Location/Qualifiers
     source          1..7299
                     /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             2457..2474
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2526..2581
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2580..2599)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4817..4835)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4903..5007
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5008..5076,5077..5868)
                     /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(5226..5245)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6039..6627
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6528..6547
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6781..6798
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6915..6936
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6951..6956,6957..6974,6975..6981)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6989..7005
                     /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     6994..7016
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7013..7029
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7013..7029
                     /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 gccaccgcca
     1201 gcatcaccgt gaccgtgctg atcaccgagt gcctgttcag cagccggcct ccccctagcc
     1261 ccccacctca tctgaaaggc gcctggcagc tgatgaaggc cacagtgacc cggcagctgt
     1321 ggtccagagg cctgagactg cccagagtgt ccagcgccgt gcccgccaac ctgagccact
     1381 gggatggcct gcctcggacc atgacaggcg ccgctgcttc tgccgctcct gccgccagag
     1441 ccagaaccac agccagcctg accagcccct tcatctgcct gcggcggttc ttctggaacc
     1501 ccttcgccag cggcaccaga ggcagcgtgt ccgccaccag agactccgtg cagcggctga
     1561 acgccgagca ggacgcctct atcgccgaga aggcccaccg ggacgccgac ctggaaaacg
     1621 agcagctgct gggcgcctgc ctggatctgc tggacatgtg ctccggcaac gccgctggca
     1681 gcgcccctag ctctgccctg gatgagctgg cccgggaccg gatctgggag ctggtggacg
     1741 tgctgctgca cgatcaccac agagcccccg ctgagcagtt cgacctggtg tacaccgccc
     1801 tgtgcatgcc agccgcccag gaccacagac aggtgcagag aagcctgctg ctggtgctgc
     1861 ggagcgtggt gcccgagaca ctgtaccggg tgttcgagag cgtggacctg ttcgtcgtgc
     1921 gggacgacga gcagtccctg caccagcgcg acgccctcat gaagttcgcc cacgcccagc
     1981 tgggagatct gcacgtgccc gacggactgg ccgaggaaga ggtgctgagc gtctacgtgg
     2041 acgacatcaa gaccgccttc cccgtgctgg ccacctgtcc cgcttggcag gtcgtggaaa
     2101 gggacgccac cacaatcgcc ctgaagtcca agctgttcgc cctgctgggg cggctgtgcg
     2161 ccgagttcga tcaggaaaag accggcaaag tcaagctggc cgatctgaga agcaccgccg
     2221 agcgggtgct gggcaccgat caggcctcta gactgctgga aggcgcccag gtggacaagg
     2281 acggcaagat cgcctatgcc cagctcgtgg ctctgctgtc ccggccacct cccaagaaga
     2341 gggcccctgc cgaagtgaca ggcgcgcccg gcagcctgca ccacatcctg gacgcccaga
     2401 agatgctgtg gaaccacagg gacaggaacc tgcctcctct ggcccctctg ggcccccacc
     2461 accatcacca ccattgagtc tagacacgtg attaattaag gatcccccga cctcgacctc
     2521 tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc
     2581 actcggaagg acatatggga gggcaaatca tttggtcgag atccctcgga gatctctagc
     2641 tagaggatcg atccccgccc cggacgaact aaacctgact acgacatctc tgccccttct
     2701 tcgcggggca gtgcatgtaa tcccttcagt tggttggtac aacttgccaa ctgggccctg
     2761 ttccacatgt gacacggggg gggaccaaac acaaaggggt tctctgactg tagttgacat
     2821 ccttataaat ggatgtgcac atttgccaac actgagtggc tttcatcctg gagcagactt
     2881 tgcagtctgt ggactgcaac acaacattgc ctttatgtgt aactcttggc tgaagctctt
     2941 acaccaatgc tgggggacat gtacctccca ggggcccagg aagactacgg gaggctacac
     3001 caacgtcaat cagaggggcc tgtgtagcta ccgataagcg gaccctcaag agggcattag
     3061 caatagtgtt tataaggccc ccttgttaac cctaaacggg tagcatatgc ttcccgggta
     3121 gtagtatata ctatccagac taaccctaat tcaatagcat atgttaccca acgggaagca
     3181 tatgctatcg aattagggtt agtaaaaggg tcctaaggaa cagcgatatc tcccacccca
     3241 tgagctgtca cggttttatt tacatggggt caggattcca cgagggtagt gaaccatttt
     3301 agtcacaagg gcagtggctg aagatcaagg agcgggcagt gaactctcct gaatcttcgc
     3361 ctgcttcttc attctccttc gtttagctaa tagaataact gctgagttgt gaacagtaag
     3421 gtgtatgtga ggtgctcgaa aacaaggttt caggtgacgc ccccagaata aaatttggac
     3481 ggggggttca gtggtggcat tgtgctatga caccaatata accctcacaa accccttggg
     3541 caataaatac tagtgtagga atgaaacatt ctgaatatct ttaacaatag aaatccatgg
     3601 ggtggggaca agccgtaaag actggatgtc catctcacac gaatttatgg ctatgggcaa
     3661 cacataatcc tagtgcaata tgatactggg gttattaaga tgtgtcccag gcagggacca
     3721 agacaggtga accatgttgt tacactctat ttgtaacaag gggaaagaga gtggacgccg
     3781 acagcagcgg actccactgg ttgtctctaa cacccccgaa aattaaacgg ggctccacgc
     3841 caatggggcc cataaacaaa gacaagtggc cactcttttt tttgaaattg tggagtgggg
     3901 gcacgcgtca gcccccacac gccgccctgc ggttttggac tgtaaaataa gggtgtaata
     3961 acttggctga ttgtaacccc gctaaccact gcggtcaaac cacttgccca caaaaccact
     4021 aatggcaccc cggggaatac ctgcataagt aggtgggcgg gccaagatag gggcgcgatt
     4081 gctgcgatct ggaggacaaa ttacacacac ttgcgcctga gcgccaagca cagggttgtt
     4141 ggtcctcata ttcacgaggt cgctgagagc acggtgggct aatgttgcca tgggtagcat
     4201 atactaccca aatatctgga tagcatatgc tatcctaatc tatatctggg tagcataggc
     4261 tatcctaatc tatatctggg tagcatatgc tatcctaatc tatatctggg tagtatatgc
     4321 tatcctaatt tatatctggg tagcataggc tatcctaatc tatatctggg tagcatatgc
     4381 tatcctaatc tatatctggg tagtatatgc tatcctaatc tgtatccggg tagcatatgc
     4441 tatcctaata gagattaggg tagtatatgc tatcctaatt tatatctggg tagcatatac
     4501 tacccaaata tctggatagc atatgctatc ctaatctata tctgggtagc atatgctatc
     4561 ctaatctata tctgggtagc ataggctatc ctaatctata tctgggtagc atatgctatc
     4621 ctaatctata tctgggtagt atatgctatc ctaatttata tctgggtagc ataggctatc
     4681 ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt atatgctatc
     4741 ctaatctgta tccgggtagc atatgctatc ctcatgataa gctgtcaaac atgagaattt
     4801 tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     4861 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     4921 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     4981 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     5041 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     5101 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     5161 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     5221 gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc
     5281 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     5341 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     5401 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     5461 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     5521 ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta
     5581 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     5641 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     5701 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     5761 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     5821 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     5881 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     5941 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     6001 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     6061 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     6121 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     6181 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     6241 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     6301 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     6361 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     6421 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     6481 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     6541 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     6601 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     6661 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     6721 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     6781 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     6841 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     6901 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
     6961 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     7021 agctatgacc atgattacgc caagctctag ctagaggtcg accaattctc atgtttgaca
     7081 gcttatcatc gcagatccgg gcaacgttgt tgccattgct gcaggcgcag aactggtagg
     7141 tatggaagat ctatacattg aatcaatatt ggcaattagc catattagtc attggttata
     7201 tagcataaat caatattggc tattggccat tgcatacgtt gtatctatat cataatatgt
     7261 acatttatat tggctcatgt ccaatatgac cgccatgtt
//
