LOCUS       sequence-274522-        8082 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 8082)
  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 8082)
  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: D6_Ld_100150
FEATURES             Location/Qualifiers
     source          1..8082
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     201..225
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     706..725
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      748..757
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2860..2877
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2929..2984
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2983..3002)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5220..5238)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5306..5410
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5411..5479,5480..6271)
                     /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(5629..5648)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6442..7030
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6931..6950
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7184..7201
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7318..7339
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7354..7359,7360..7377,7378..7384)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7392..7408
                     /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     7397..7419
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7416..7432
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7416..7432
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7703..8082
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta caccaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcctcac tctcttccgc
      241 atcgctgtct gcgagggcca gctgttgggc tcgcggttga ggacaaactc ttcgcggtct
      301 ttccagtact cttggatcgg aaacccgtcg gcctccgaac ggtactccgc caccgaggga
      361 cctgagcgag tccgcatcga ccggatcgga aaacctctcg agaaaggcgt ctaaccagtc
      421 acagtcgcaa ggtaggctga gcaccgtggc gggcggcagc gggtggcggt cggggttgtt
      481 tctggcggag gtgctgctga tgatgtaatt aaagtaggcg gtcttgagac ggcggatggt
      541 cgaggtgagg tgtggcaggc ttgagatcca gctgttgggg tgagtactcc ctctcaaaag
      601 cgggcattac ttctgcgcta agattgtcag tttccaaaaa cgaggaggat ttgatattca
      661 cctggcccga tctggccata cacttgagtg acaatgacat ccactttgcc tttctctcca
      721 caggtgtcca ctcccaggtc caagtttgcc accatggagt ttcagaccca ggtactcatg
      781 tccctgctgc tctgcatgtc tggtgcggcc gccaccaacc ctctgcagag cagacacatg
      841 cagctgaccc agaacgccgt gaagaagtcc gaggccaacc tggtcaactg ccgggaagtg
      901 aacgccgacc tgaagtccgg cggaggcgtg aacgccgccc aggccattgc cgagatgcgg
      961 cggcagcgcg aggaactgat gaacatcgtg gccctggaac gcgagcgggt ggtgtccgct
     1021 agaagcctgc tgcaggtctg cgaggacgag ctggccagcg acctgagcgt gctgttcggc
     1081 gtggccgacc acaacttcac cgcccggatc agaagcctgg aagagaagcg gaagcacctg
     1141 gaaggcctgc acagcatgct gaacaccacc cccttcggcg ccgtggaact gcggcggtct
     1201 agcgagatta gagccctgca ggccgccctg ctgcacgaga tgcgggccag caagaagaaa
     1261 gccgagaacg gcgtggccaa cggcgaggcc tgcaccaaga ccagcgacaa gtacagcgtg
     1321 gtgttcgaca tcggcagcac cggcaaccgg gtgcacgtgt acaagtaccg ggtggcccct
     1381 gccacccata cagccgccgc tgccggctcc gagctgagcg acatcgacct ggtggaagaa
     1441 ctgttcgagc tgaaccacaa ggccctgagc gagctggaca accccgtgca ggatgcccct
     1501 gaggccctgt gggagctgtt catgaaggcc aagtacttcg tgcccgccga gctgcacgcc
     1561 tgcaccgccg tggaattcaa ggccacagcc ggcctgcgga tgctgggcat ggaaaaggcc
     1621 accgagatcc tggacggcat ccgggccaga taccggaacg agacattctg gctgcggggc
     1681 aacgcccccg tgcggatcct ggatgcctgc gaagagggcc ccatggcctg gctgaccgtg
     1741 aactacctgc tgggcgtgtt cagccggggc accaaggcca ctgccagcac cgtggccgtg
     1801 atcgatctgg gcggaggcag cacccagatc gtgttcgagc ctggcgagag cgccttccac
     1861 ggcatgcgga ccgacctgag atacagcgcc accctgggca gcagaagcgt gtccgcctac
     1921 cagcacagct acgagggcta tggcctgcac gccgccacaa aagagctgct gttccatatt
     1981 cagggcaaga gccaggaaaa gcctggcggc ggaaccacca ccagcaccgc caccaccacc
     2041 acaaccacta cccccgccaa cagcggcgat aaggccctgc ccgtgtggaa cgtgctgggc
     2101 aatctgggcg ccgacggcag cagcgagagg gacggaatcg tgaccaaacg ggcccctccc
     2161 atgccccctc cacctctgcc tgatgccgag gccgtggaag ccttcccttg cttcgccgtg
     2221 ggctacgagg accccctggg cgtgaagaac atcaagaaga acaacgccgg cgagcccgcc
     2281 atgcccccca atttccaggc ctgcgccaac ctgttccggg accggctgct gaagcctgtg
     2341 ggcctgacat gcgaggccgc caactgtgga atcgccggcg tgatgcagcc ccccctgacc
     2401 aatttcaccg gcgagatcta cgtgttcagc ttcatcttcg acctgctggc cctggccaac
     2461 agcagcctgg tgcctgctgg cgccgctgtg tcccgcgaga agttcgaagt gaagctgccc
     2521 gacctggcca caatcgccga gggccactgt gccgccttca gcctgaccag aattgccgag
     2581 gccaccgcca aagagggcct gggcagcctg aagcccgagt acgagtgcat gtactacagc
     2641 tacgtgtacg ccctgctgag atacggctac gaggtgcccg aggaccgggt gctgcacgtg
     2701 gtcaagaaga tccggggcta cgagacagcc tggtccctgg gcgccagcct gctgagcctg
     2761 acaggcgcgc ccggcagcct gcaccacatc ctggacgccc agaagatgct gtggaaccac
     2821 agggacagga acctgcctcc tctggcccct ctgggccccc accaccatca ccaccattga
     2881 gtctagacac gtgattaatt aaggatcccc cgacctcgac ctctggctaa taaaggaaat
     2941 ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg
     3001 ggagggcaaa tcatttggtc gagatccctc ggagatctct agctagagga tcgatccccg
     3061 ccccggacga actaaacctg actacgacat ctctgcccct tcttcgcggg gcagtgcatg
     3121 taatcccttc agttggttgg tacaacttgc caactgggcc ctgttccaca tgtgacacgg
     3181 ggggggacca aacacaaagg ggttctctga ctgtagttga catccttata aatggatgtg
     3241 cacatttgcc aacactgagt ggctttcatc ctggagcaga ctttgcagtc tgtggactgc
     3301 aacacaacat tgcctttatg tgtaactctt ggctgaagct cttacaccaa tgctggggga
     3361 catgtacctc ccaggggccc aggaagacta cgggaggcta caccaacgtc aatcagaggg
     3421 gcctgtgtag ctaccgataa gcggaccctc aagagggcat tagcaatagt gtttataagg
     3481 cccccttgtt aaccctaaac gggtagcata tgcttcccgg gtagtagtat atactatcca
     3541 gactaaccct aattcaatag catatgttac ccaacgggaa gcatatgcta tcgaattagg
     3601 gttagtaaaa gggtcctaag gaacagcgat atctcccacc ccatgagctg tcacggtttt
     3661 atttacatgg ggtcaggatt ccacgagggt agtgaaccat tttagtcaca agggcagtgg
     3721 ctgaagatca aggagcgggc agtgaactct cctgaatctt cgcctgcttc ttcattctcc
     3781 ttcgtttagc taatagaata actgctgagt tgtgaacagt aaggtgtatg tgaggtgctc
     3841 gaaaacaagg tttcaggtga cgcccccaga ataaaatttg gacggggggt tcagtggtgg
     3901 cattgtgcta tgacaccaat ataaccctca caaacccctt gggcaataaa tactagtgta
     3961 ggaatgaaac attctgaata tctttaacaa tagaaatcca tggggtgggg acaagccgta
     4021 aagactggat gtccatctca cacgaattta tggctatggg caacacataa tcctagtgca
     4081 atatgatact ggggttatta agatgtgtcc caggcaggga ccaagacagg tgaaccatgt
     4141 tgttacactc tatttgtaac aaggggaaag agagtggacg ccgacagcag cggactccac
     4201 tggttgtctc taacaccccc gaaaattaaa cggggctcca cgccaatggg gcccataaac
     4261 aaagacaagt ggccactctt ttttttgaaa ttgtggagtg ggggcacgcg tcagccccca
     4321 cacgccgccc tgcggttttg gactgtaaaa taagggtgta ataacttggc tgattgtaac
     4381 cccgctaacc actgcggtca aaccacttgc ccacaaaacc actaatggca ccccggggaa
     4441 tacctgcata agtaggtggg cgggccaaga taggggcgcg attgctgcga tctggaggac
     4501 aaattacaca cacttgcgcc tgagcgccaa gcacagggtt gttggtcctc atattcacga
     4561 ggtcgctgag agcacggtgg gctaatgttg ccatgggtag catatactac ccaaatatct
     4621 ggatagcata tgctatccta atctatatct gggtagcata ggctatccta atctatatct
     4681 gggtagcata tgctatccta atctatatct gggtagtata tgctatccta atttatatct
     4741 gggtagcata ggctatccta atctatatct gggtagcata tgctatccta atctatatct
     4801 gggtagtata tgctatccta atctgtatcc gggtagcata tgctatccta atagagatta
     4861 gggtagtata tgctatccta atttatatct gggtagcata tactacccaa atatctggat
     4921 agcatatgct atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt
     4981 agcataggct atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt
     5041 agtatatgct atcctaattt atatctgggt agcataggct atcctaatct atatctgggt
     5101 agcatatgct atcctaatct atatctgggt agtatatgct atcctaatct gtatccgggt
     5161 agcatatgct atcctcatga taagctgtca aacatgagaa ttttcttgaa gacgaaaggg
     5221 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     5281 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     5341 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     5401 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     5461 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     5521 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     5581 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     5641 ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     5701 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     5761 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     5821 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     5881 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     5941 caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     6001 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     6061 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     6121 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     6181 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     6241 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     6301 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     6361 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     6421 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     6481 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     6541 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     6601 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     6661 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     6721 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     6781 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6841 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6901 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     6961 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     7021 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     7081 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     7141 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     7201 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     7261 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     7321 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     7381 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     7441 cgccaagctc tagctagagg tcgaccaatt ctcatgtttg acagcttatc atcgcagatc
     7501 cgggcaacgt tgttgccatt gctgcaggcg cagaactggt aggtatggaa gatctataca
     7561 ttgaatcaat attggcaatt agccatatta gtcattggtt atatagcata aatcaatatt
     7621 ggctattggc cattgcatac gttgtatcta tatcataata tgtacattta tattggctca
     7681 tgtccaatat gaccgccatg ttgacattga ttattgacta gttattaata gtaatcaatt
     7741 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     7801 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     7861 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     7921 actgcccact tggcagtaca tcaagtgtat catatgccaa gtccgccccc tattgacgtc
     7981 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttacg ggactttcct
     8041 acttggcagt acatctacgt attagtcatc gctattacca tg
//
