LOCUS       sequence-274855-        6972 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 6972)
  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 6972)
  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: D74_Ld_291730
FEATURES             Location/Qualifiers
     source          1..6972
                     /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             2130..2147
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2199..2254
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2253..2272)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4490..4508)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4576..4680
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4681..4749,4750..5541)
                     /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(4899..4918)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5712..6300
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6201..6220
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6454..6471
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6588..6609
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6624..6629,6630..6647,6648..6654)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6662..6678
                     /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     6667..6689
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6686..6702
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6686..6702
                     /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 gccggagcta
     1201 cacccgcatc cgccttttac gtacctggtg ccgccgaaaa ggcctataag aagggcgaag
     1261 aggtgaaatt tatggttaac agcctgcgct caagtagcga aatgttccca atagactata
     1321 gcaaaatgcc attctgccag ccagctcgcc aagaattcaa agaggagtct attggggaga
     1381 ttatttgggg tgaccggatg cttaattccc tgtataccgt gcgaatgaaa gaggacgtca
     1441 agtgcatggc tctgcccgac tgtgacttca ttgctaacac agagactata cgccgtaagg
     1501 aaagtaagaa cctgacaaag atgattaata agtggtaccg cgtttatatg aatattgata
     1561 acctgccagt attcagtaca aatccggaaa gtacccagat gtctgagtgc gccaagaagc
     1621 tgggtaaaga tgtaaagatc tacgcacagc gaggatttcc attgggcgtg cccgcgaagt
     1681 gcacaagtga cagagccgct ctgctgaaca accacttgga cttcactatt cattacaacc
     1741 acgactccca aacgacccga actaccgccg aagaggaacg gcgatatatt gtcgtcttca
     1801 tcgatgtaaa agctcgtagc atagcctggt cagactcact cgaatgcaat tctcagatga
     1861 aagtcgcacc cgaggtgttg gcacccatgc gcgggctcaa aatgaagaat gtgactcaga
     1921 ataaggccac agtctactgg acctactccg taaagtggaa agagaaccca aacgtcaaat
     1981 gggctacacg ttgggacttt tacctgacag cagcagcagc tgcagccccg gccggcgcgc
     2041 ccggcagcct gcaccacatc ctggacgccc agaagatgct gtggaaccac agggacagga
     2101 acctgcctcc tctggcccct ctgggccccc accaccatca ccaccattga gtctagacac
     2161 gtgattaatt aaggatcccc cgacctcgac ctctggctaa taaaggaaat ttattttcat
     2221 tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa
     2281 tcatttggtc gagatccctc ggagatctct agctagagga tcgatccccg ccccggacga
     2341 actaaacctg actacgacat ctctgcccct tcttcgcggg gcagtgcatg taatcccttc
     2401 agttggttgg tacaacttgc caactgggcc ctgttccaca tgtgacacgg ggggggacca
     2461 aacacaaagg ggttctctga ctgtagttga catccttata aatggatgtg cacatttgcc
     2521 aacactgagt ggctttcatc ctggagcaga ctttgcagtc tgtggactgc aacacaacat
     2581 tgcctttatg tgtaactctt ggctgaagct cttacaccaa tgctggggga catgtacctc
     2641 ccaggggccc aggaagacta cgggaggcta caccaacgtc aatcagaggg gcctgtgtag
     2701 ctaccgataa gcggaccctc aagagggcat tagcaatagt gtttataagg cccccttgtt
     2761 aaccctaaac gggtagcata tgcttcccgg gtagtagtat atactatcca gactaaccct
     2821 aattcaatag catatgttac ccaacgggaa gcatatgcta tcgaattagg gttagtaaaa
     2881 gggtcctaag gaacagcgat atctcccacc ccatgagctg tcacggtttt atttacatgg
     2941 ggtcaggatt ccacgagggt agtgaaccat tttagtcaca agggcagtgg ctgaagatca
     3001 aggagcgggc agtgaactct cctgaatctt cgcctgcttc ttcattctcc ttcgtttagc
     3061 taatagaata actgctgagt tgtgaacagt aaggtgtatg tgaggtgctc gaaaacaagg
     3121 tttcaggtga cgcccccaga ataaaatttg gacggggggt tcagtggtgg cattgtgcta
     3181 tgacaccaat ataaccctca caaacccctt gggcaataaa tactagtgta ggaatgaaac
     3241 attctgaata tctttaacaa tagaaatcca tggggtgggg acaagccgta aagactggat
     3301 gtccatctca cacgaattta tggctatggg caacacataa tcctagtgca atatgatact
     3361 ggggttatta agatgtgtcc caggcaggga ccaagacagg tgaaccatgt tgttacactc
     3421 tatttgtaac aaggggaaag agagtggacg ccgacagcag cggactccac tggttgtctc
     3481 taacaccccc gaaaattaaa cggggctcca cgccaatggg gcccataaac aaagacaagt
     3541 ggccactctt ttttttgaaa ttgtggagtg ggggcacgcg tcagccccca cacgccgccc
     3601 tgcggttttg gactgtaaaa taagggtgta ataacttggc tgattgtaac cccgctaacc
     3661 actgcggtca aaccacttgc ccacaaaacc actaatggca ccccggggaa tacctgcata
     3721 agtaggtggg cgggccaaga taggggcgcg attgctgcga tctggaggac aaattacaca
     3781 cacttgcgcc tgagcgccaa gcacagggtt gttggtcctc atattcacga ggtcgctgag
     3841 agcacggtgg gctaatgttg ccatgggtag catatactac ccaaatatct ggatagcata
     3901 tgctatccta atctatatct gggtagcata ggctatccta atctatatct gggtagcata
     3961 tgctatccta atctatatct gggtagtata tgctatccta atttatatct gggtagcata
     4021 ggctatccta atctatatct gggtagcata tgctatccta atctatatct gggtagtata
     4081 tgctatccta atctgtatcc gggtagcata tgctatccta atagagatta gggtagtata
     4141 tgctatccta atttatatct gggtagcata tactacccaa atatctggat agcatatgct
     4201 atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt agcataggct
     4261 atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt agtatatgct
     4321 atcctaattt atatctgggt agcataggct atcctaatct atatctgggt agcatatgct
     4381 atcctaatct atatctgggt agtatatgct atcctaatct gtatccgggt agcatatgct
     4441 atcctcatga taagctgtca aacatgagaa ttttcttgaa gacgaaaggg cctcgtgata
     4501 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     4561 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     4621 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     4681 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     4741 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     4801 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     4861 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     4921 cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     4981 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     5041 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     5101 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     5161 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     5221 cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     5281 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     5341 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     5401 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     5461 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     5521 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     5581 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     5641 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     5701 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     5761 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     5821 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     5881 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     5941 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     6001 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     6061 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     6121 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     6181 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     6241 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     6301 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     6361 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     6421 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     6481 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     6541 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     6601 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     6661 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagctc
     6721 tagctagagg tcgaccaatt ctcatgtttg acagcttatc atcgcagatc cgggcaacgt
     6781 tgttgccatt gctgcaggcg cagaactggt aggtatggaa gatctataca ttgaatcaat
     6841 attggcaatt agccatatta gtcattggtt atatagcata aatcaatatt ggctattggc
     6901 cattgcatac gttgtatcta tatcataata tgtacattta tattggctca tgtccaatat
     6961 gaccgccatg tt
//
