LOCUS       sequence-274721-        7779 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 7779)
  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 7779)
  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: D48_Ld_353820
FEATURES             Location/Qualifiers
     source          1..7779
                     /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             2937..2954
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3006..3061
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3060..3079)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5297..5315)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5383..5487
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5488..5556,5557..6348)
                     /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(5706..5725)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6519..7107
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7008..7027
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7261..7278
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7395..7416
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7431..7436,7437..7454,7455..7461)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7469..7485
                     /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     7474..7496
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7493..7509
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7493..7509
                     /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 gccaccgtga
     1201 acctgctgcc tctgatgggc gagggcctgt tcgacgtgga agtcgtgtgc cgcagcgcca
     1261 tgcccagcga gcccaccgtg tacgacagaa gcgtgatcag cgccttccgg ttcagcgtga
     1321 acgccagcgc cgacagcacc aaagaaaaag tgcatctgcc cctgtacgtg gaacggatcc
     1381 tggacagcga ctacgtgcgg acctactaca tcaacagcac cgacaagtac agcatcagcg
     1441 acagcaaggg ctgcgtgcac accaagggac tgtacgagag cagcgacgag gtgctgctga
     1501 gcgagagcct gatcaccatg ctgcggagcc ccctgagcgc cacccccagc cctagcatga
     1561 tcagaggaat cgccgccgac aactactcca gccagtacaa cctgaccatc cccagcagct
     1621 acggcggcct gggcagccct gccacatata gagccgccgt gctgacctcc acccagcact
     1681 ggctggtcaa cctgatgcag gtcgacctca cccccgtgtc catcgaagtg atgaccaccg
     1741 acagcgcctc caagatcaac aactgcctgt tcaccttcag ctttttcggc agcaatgtgt
     1801 ctgccgtgcg gggctccctg cccagctcct gcaagaacag cagctccgag caccccagag
     1861 tgtggaaaag cagcaacgtg gccctgccta tgatgcgggc cctggctggc gccgctaagg
     1921 ccttcagcag cagacccagc gtgttcagct ccagcagcga gacaagctcc tccaccagcg
     1981 ctgccgacac cgccaactac accatgcccg ccttccccga cgactttacc gccaatttcc
     2041 tggtggtgtc ccccctgaag aagtccttct accaggtccg agagagctac agcctgtacg
     2101 ccggcttcag cctgaccagc ctgcagttcc ctctgaaggg cattgccggc agagtgtaca
     2161 agcacgagtg gtacaccaac agctggaacc agatgagcta ctaccacacc gagttcgccg
     2221 tgccctccgg ccaggaaatg gccgacgagg ccctgcggaa gtacttcttc cctgacctga
     2281 acacctgtcg gcgggtcgtg atcgcctacg acctgatggc ccggtctagc agctctctgc
     2341 tgctgtacag ccccaacgtg ccccccacct tcatcggcaa ccagaccgtg cggaacatcc
     2401 cctgcggcgt gtgggtggcc gagatccagg gcgtcagagt gacctggtac tgggccacca
     2461 ccgatctggt ggataccgcc agcttctgga ccgtggacag catcgagagc ggcgcctccg
     2521 cctacaccag gctcgtgcgg atgacagtga ccggcaaagg cggcgctccc cccctgtttg
     2581 cccaccaccc attctttcca cagtcctacg cctttcccgt ggacgaccgg gatgtggcct
     2641 gcaccgccat gatgcctagc gaggccgaca tgggctgcta cagctacgcc aagaacgccg
     2701 attacaccta catctacgag atcaccagct tcgtgcccta tgtgcggcgg aacgactacc
     2761 ggctgcctgc cgcctgtgtg ggcgtgaaga tcctgggctc catccctagc ttccagtgca
     2821 actacaaggg catcaccgga ggcgcgcccg gcagcctgca ccacatcctg gacgcccaga
     2881 agatgctgtg gaaccacagg gacaggaacc tgcctcctct ggcccctctg ggcccccacc
     2941 accatcacca ccattgagtc tagacacgtg attaattaag gatcccccga cctcgacctc
     3001 tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc
     3061 actcggaagg acatatggga gggcaaatca tttggtcgag atccctcgga gatctctagc
     3121 tagaggatcg atccccgccc cggacgaact aaacctgact acgacatctc tgccccttct
     3181 tcgcggggca gtgcatgtaa tcccttcagt tggttggtac aacttgccaa ctgggccctg
     3241 ttccacatgt gacacggggg gggaccaaac acaaaggggt tctctgactg tagttgacat
     3301 ccttataaat ggatgtgcac atttgccaac actgagtggc tttcatcctg gagcagactt
     3361 tgcagtctgt ggactgcaac acaacattgc ctttatgtgt aactcttggc tgaagctctt
     3421 acaccaatgc tgggggacat gtacctccca ggggcccagg aagactacgg gaggctacac
     3481 caacgtcaat cagaggggcc tgtgtagcta ccgataagcg gaccctcaag agggcattag
     3541 caatagtgtt tataaggccc ccttgttaac cctaaacggg tagcatatgc ttcccgggta
     3601 gtagtatata ctatccagac taaccctaat tcaatagcat atgttaccca acgggaagca
     3661 tatgctatcg aattagggtt agtaaaaggg tcctaaggaa cagcgatatc tcccacccca
     3721 tgagctgtca cggttttatt tacatggggt caggattcca cgagggtagt gaaccatttt
     3781 agtcacaagg gcagtggctg aagatcaagg agcgggcagt gaactctcct gaatcttcgc
     3841 ctgcttcttc attctccttc gtttagctaa tagaataact gctgagttgt gaacagtaag
     3901 gtgtatgtga ggtgctcgaa aacaaggttt caggtgacgc ccccagaata aaatttggac
     3961 ggggggttca gtggtggcat tgtgctatga caccaatata accctcacaa accccttggg
     4021 caataaatac tagtgtagga atgaaacatt ctgaatatct ttaacaatag aaatccatgg
     4081 ggtggggaca agccgtaaag actggatgtc catctcacac gaatttatgg ctatgggcaa
     4141 cacataatcc tagtgcaata tgatactggg gttattaaga tgtgtcccag gcagggacca
     4201 agacaggtga accatgttgt tacactctat ttgtaacaag gggaaagaga gtggacgccg
     4261 acagcagcgg actccactgg ttgtctctaa cacccccgaa aattaaacgg ggctccacgc
     4321 caatggggcc cataaacaaa gacaagtggc cactcttttt tttgaaattg tggagtgggg
     4381 gcacgcgtca gcccccacac gccgccctgc ggttttggac tgtaaaataa gggtgtaata
     4441 acttggctga ttgtaacccc gctaaccact gcggtcaaac cacttgccca caaaaccact
     4501 aatggcaccc cggggaatac ctgcataagt aggtgggcgg gccaagatag gggcgcgatt
     4561 gctgcgatct ggaggacaaa ttacacacac ttgcgcctga gcgccaagca cagggttgtt
     4621 ggtcctcata ttcacgaggt cgctgagagc acggtgggct aatgttgcca tgggtagcat
     4681 atactaccca aatatctgga tagcatatgc tatcctaatc tatatctggg tagcataggc
     4741 tatcctaatc tatatctggg tagcatatgc tatcctaatc tatatctggg tagtatatgc
     4801 tatcctaatt tatatctggg tagcataggc tatcctaatc tatatctggg tagcatatgc
     4861 tatcctaatc tatatctggg tagtatatgc tatcctaatc tgtatccggg tagcatatgc
     4921 tatcctaata gagattaggg tagtatatgc tatcctaatt tatatctggg tagcatatac
     4981 tacccaaata tctggatagc atatgctatc ctaatctata tctgggtagc atatgctatc
     5041 ctaatctata tctgggtagc ataggctatc ctaatctata tctgggtagc atatgctatc
     5101 ctaatctata tctgggtagt atatgctatc ctaatttata tctgggtagc ataggctatc
     5161 ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt atatgctatc
     5221 ctaatctgta tccgggtagc atatgctatc ctcatgataa gctgtcaaac atgagaattt
     5281 tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     5341 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     5401 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     5461 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     5521 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     5581 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     5641 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     5701 gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc
     5761 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     5821 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     5881 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     5941 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     6001 ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta
     6061 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     6121 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     6181 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     6241 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     6301 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     6361 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     6421 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     6481 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     6541 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     6601 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     6661 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     6721 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     6781 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     6841 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     6901 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     6961 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     7021 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     7081 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     7141 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     7201 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     7261 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     7321 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     7381 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
     7441 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     7501 agctatgacc atgattacgc caagctctag ctagaggtcg accaattctc atgtttgaca
     7561 gcttatcatc gcagatccgg gcaacgttgt tgccattgct gcaggcgcag aactggtagg
     7621 tatggaagat ctatacattg aatcaatatt ggcaattagc catattagtc attggttata
     7681 tagcataaat caatattggc tattggccat tgcatacgtt gtatctatat cataatatgt
     7741 acatttatat tggctcatgt ccaatatgac cgccatgtt
//
