LOCUS       sequence-274747-        6342 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 6342)
  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 6342)
  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: D79_Ld_366740
FEATURES             Location/Qualifiers
     source          1..6342
                     /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             1120..1137
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1189..1244
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1243..1262)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3480..3498)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3566..3670
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3671..3739,3740..4531)
                     /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(3889..3908)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4702..5290
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5191..5210
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5444..5461
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5578..5599
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5614..5619,5620..5637,5638..5644)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5652..5668
                     /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     5657..5679
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5676..5692
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5676..5692
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        5963..6342
                     /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 gccagactgg ttgttagaat ggtgcaggtc
      841 gtgcacagac acggcgccag atctgccctg atcaacgaca ataccaccga gatctgcggc
      901 acactgtacc cttgtggcga actgacaggc gaaggcgtgg aaatggttcg agccatcgga
      961 gagtctcccc atctgcaccc ttttgccgtg actcaggatt ggtctagcct gcctcctctg
     1021 acaggcgcgc ccggcagcct gcaccacatc ctggacgccc agaagatgct gtggaaccac
     1081 agggacagga acctgcctcc tctggcccct ctgggccccc accaccatca ccaccattga
     1141 gtctagacac gtgattaatt aaggatcccc cgacctcgac ctctggctaa taaaggaaat
     1201 ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg
     1261 ggagggcaaa tcatttggtc gagatccctc ggagatctct agctagagga tcgatccccg
     1321 ccccggacga actaaacctg actacgacat ctctgcccct tcttcgcggg gcagtgcatg
     1381 taatcccttc agttggttgg tacaacttgc caactgggcc ctgttccaca tgtgacacgg
     1441 ggggggacca aacacaaagg ggttctctga ctgtagttga catccttata aatggatgtg
     1501 cacatttgcc aacactgagt ggctttcatc ctggagcaga ctttgcagtc tgtggactgc
     1561 aacacaacat tgcctttatg tgtaactctt ggctgaagct cttacaccaa tgctggggga
     1621 catgtacctc ccaggggccc aggaagacta cgggaggcta caccaacgtc aatcagaggg
     1681 gcctgtgtag ctaccgataa gcggaccctc aagagggcat tagcaatagt gtttataagg
     1741 cccccttgtt aaccctaaac gggtagcata tgcttcccgg gtagtagtat atactatcca
     1801 gactaaccct aattcaatag catatgttac ccaacgggaa gcatatgcta tcgaattagg
     1861 gttagtaaaa gggtcctaag gaacagcgat atctcccacc ccatgagctg tcacggtttt
     1921 atttacatgg ggtcaggatt ccacgagggt agtgaaccat tttagtcaca agggcagtgg
     1981 ctgaagatca aggagcgggc agtgaactct cctgaatctt cgcctgcttc ttcattctcc
     2041 ttcgtttagc taatagaata actgctgagt tgtgaacagt aaggtgtatg tgaggtgctc
     2101 gaaaacaagg tttcaggtga cgcccccaga ataaaatttg gacggggggt tcagtggtgg
     2161 cattgtgcta tgacaccaat ataaccctca caaacccctt gggcaataaa tactagtgta
     2221 ggaatgaaac attctgaata tctttaacaa tagaaatcca tggggtgggg acaagccgta
     2281 aagactggat gtccatctca cacgaattta tggctatggg caacacataa tcctagtgca
     2341 atatgatact ggggttatta agatgtgtcc caggcaggga ccaagacagg tgaaccatgt
     2401 tgttacactc tatttgtaac aaggggaaag agagtggacg ccgacagcag cggactccac
     2461 tggttgtctc taacaccccc gaaaattaaa cggggctcca cgccaatggg gcccataaac
     2521 aaagacaagt ggccactctt ttttttgaaa ttgtggagtg ggggcacgcg tcagccccca
     2581 cacgccgccc tgcggttttg gactgtaaaa taagggtgta ataacttggc tgattgtaac
     2641 cccgctaacc actgcggtca aaccacttgc ccacaaaacc actaatggca ccccggggaa
     2701 tacctgcata agtaggtggg cgggccaaga taggggcgcg attgctgcga tctggaggac
     2761 aaattacaca cacttgcgcc tgagcgccaa gcacagggtt gttggtcctc atattcacga
     2821 ggtcgctgag agcacggtgg gctaatgttg ccatgggtag catatactac ccaaatatct
     2881 ggatagcata tgctatccta atctatatct gggtagcata ggctatccta atctatatct
     2941 gggtagcata tgctatccta atctatatct gggtagtata tgctatccta atttatatct
     3001 gggtagcata ggctatccta atctatatct gggtagcata tgctatccta atctatatct
     3061 gggtagtata tgctatccta atctgtatcc gggtagcata tgctatccta atagagatta
     3121 gggtagtata tgctatccta atttatatct gggtagcata tactacccaa atatctggat
     3181 agcatatgct atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt
     3241 agcataggct atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt
     3301 agtatatgct atcctaattt atatctgggt agcataggct atcctaatct atatctgggt
     3361 agcatatgct atcctaatct atatctgggt agtatatgct atcctaatct gtatccgggt
     3421 agcatatgct atcctcatga taagctgtca aacatgagaa ttttcttgaa gacgaaaggg
     3481 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     3541 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     3601 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     3661 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     3721 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     3781 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     3841 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     3901 ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     3961 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     4021 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     4081 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     4141 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     4201 caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     4261 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     4321 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     4381 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     4441 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     4501 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     4561 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     4621 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     4681 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     4741 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     4801 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     4861 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     4921 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     4981 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     5041 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     5101 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     5161 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     5221 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     5281 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     5341 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     5401 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     5461 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     5521 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     5581 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     5641 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     5701 cgccaagctc tagctagagg tcgaccaatt ctcatgtttg acagcttatc atcgcagatc
     5761 cgggcaacgt tgttgccatt gctgcaggcg cagaactggt aggtatggaa gatctataca
     5821 ttgaatcaat attggcaatt agccatatta gtcattggtt atatagcata aatcaatatt
     5881 ggctattggc cattgcatac gttgtatcta tatcataata tgtacattta tattggctca
     5941 tgtccaatat gaccgccatg ttgacattga ttattgacta gttattaata gtaatcaatt
     6001 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     6061 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     6121 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     6181 actgcccact tggcagtaca tcaagtgtat catatgccaa gtccgccccc tattgacgtc
     6241 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttacg ggactttcct
     6301 acttggcagt acatctacgt attagtcatc gctattacca tg
//
