LOCUS       sequence-274682-        7062 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 7062)
  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 7062)
  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: D85_Ld_120350
FEATURES             Location/Qualifiers
     source          1..7062
                     /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             2220..2237
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2289..2344
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2343..2362)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4580..4598)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4666..4770
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4771..4839,4840..5631)
                     /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(4989..5008)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5802..6390
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6291..6310
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6544..6561
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6678..6699
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6714..6719,6720..6737,6738..6744)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6752..6768
                     /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     6757..6779
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6776..6792
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6776..6792
                     /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 gcctggtggg
     1201 ataaagggca catgtgtatt gccgaaattg ctagacgtaa cttgaagcct aacgtgcagt
     1261 ccaaagtgca ggcttgtgcc gatgtactca ataagattgg accttttcct aagtctacta
     1321 acatcgtgga gcttggcccc tgggctgatg atctcaaaag catgggcctc tacaccatga
     1381 gtacatggca ctttattgat acaatttaca acccgcagga cgttaaggta accatcaatc
     1441 cagtggagat tgtaaatgtg gcgtctgtga tcccaatgct cattagcgcg ataacctctc
     1501 ctgctgcaac cagtgatatt attaccacta gtgtcgcaaa cctgatacac tttgtgggag
     1561 acattcatat gcctttgcat agtgccgacc tgttcagccc tgaataccct ttgggtgact
     1621 tgggtggaaa caagcagatc gtcatagtaa acgaatccgc aggcacctct atgaaattgc
     1681 atgccttttg ggacagcatg tgtgaggggc ctcaaaacaa tgctgtaagg cccctggaca
     1741 aagacgccta cgcagagctg tcagcctttg tggacaatct ggtgaagtca tacagtttca
     1801 ctgaggaaca gatgatgatg actaacagta ccattatggc cgccgagtcc tatgaattgg
     1861 ccgtgaagaa cgtctacccg ggtatatctg ataggacagt gctttccgaa acttacaagg
     1921 ccaacgggaa gattctggca gccgggcggg tcactctggc aggctacaga ctggctacaa
     1981 ttctgaatac agctctggct ggggtgtcat tggatacaat catgaacggg acaaagcaca
     2041 tgcaggatga ggttgaggtg acccatacag gcgacaccta taactactac gctttctcag
     2101 gcgtcgagtc tggagcagcc gccggcgcgc ccggcagcct gcaccacatc ctggacgccc
     2161 agaagatgct gtggaaccac agggacagga acctgcctcc tctggcccct ctgggccccc
     2221 accaccatca ccaccattga gtctagacac gtgattaatt aaggatcccc cgacctcgac
     2281 ctctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct
     2341 ctcactcgga aggacatatg ggagggcaaa tcatttggtc gagatccctc ggagatctct
     2401 agctagagga tcgatccccg ccccggacga actaaacctg actacgacat ctctgcccct
     2461 tcttcgcggg gcagtgcatg taatcccttc agttggttgg tacaacttgc caactgggcc
     2521 ctgttccaca tgtgacacgg ggggggacca aacacaaagg ggttctctga ctgtagttga
     2581 catccttata aatggatgtg cacatttgcc aacactgagt ggctttcatc ctggagcaga
     2641 ctttgcagtc tgtggactgc aacacaacat tgcctttatg tgtaactctt ggctgaagct
     2701 cttacaccaa tgctggggga catgtacctc ccaggggccc aggaagacta cgggaggcta
     2761 caccaacgtc aatcagaggg gcctgtgtag ctaccgataa gcggaccctc aagagggcat
     2821 tagcaatagt gtttataagg cccccttgtt aaccctaaac gggtagcata tgcttcccgg
     2881 gtagtagtat atactatcca gactaaccct aattcaatag catatgttac ccaacgggaa
     2941 gcatatgcta tcgaattagg gttagtaaaa gggtcctaag gaacagcgat atctcccacc
     3001 ccatgagctg tcacggtttt atttacatgg ggtcaggatt ccacgagggt agtgaaccat
     3061 tttagtcaca agggcagtgg ctgaagatca aggagcgggc agtgaactct cctgaatctt
     3121 cgcctgcttc ttcattctcc ttcgtttagc taatagaata actgctgagt tgtgaacagt
     3181 aaggtgtatg tgaggtgctc gaaaacaagg tttcaggtga cgcccccaga ataaaatttg
     3241 gacggggggt tcagtggtgg cattgtgcta tgacaccaat ataaccctca caaacccctt
     3301 gggcaataaa tactagtgta ggaatgaaac attctgaata tctttaacaa tagaaatcca
     3361 tggggtgggg acaagccgta aagactggat gtccatctca cacgaattta tggctatggg
     3421 caacacataa tcctagtgca atatgatact ggggttatta agatgtgtcc caggcaggga
     3481 ccaagacagg tgaaccatgt tgttacactc tatttgtaac aaggggaaag agagtggacg
     3541 ccgacagcag cggactccac tggttgtctc taacaccccc gaaaattaaa cggggctcca
     3601 cgccaatggg gcccataaac aaagacaagt ggccactctt ttttttgaaa ttgtggagtg
     3661 ggggcacgcg tcagccccca cacgccgccc tgcggttttg gactgtaaaa taagggtgta
     3721 ataacttggc tgattgtaac cccgctaacc actgcggtca aaccacttgc ccacaaaacc
     3781 actaatggca ccccggggaa tacctgcata agtaggtggg cgggccaaga taggggcgcg
     3841 attgctgcga tctggaggac aaattacaca cacttgcgcc tgagcgccaa gcacagggtt
     3901 gttggtcctc atattcacga ggtcgctgag agcacggtgg gctaatgttg ccatgggtag
     3961 catatactac ccaaatatct ggatagcata tgctatccta atctatatct gggtagcata
     4021 ggctatccta atctatatct gggtagcata tgctatccta atctatatct gggtagtata
     4081 tgctatccta atttatatct gggtagcata ggctatccta atctatatct gggtagcata
     4141 tgctatccta atctatatct gggtagtata tgctatccta atctgtatcc gggtagcata
     4201 tgctatccta atagagatta gggtagtata tgctatccta atttatatct gggtagcata
     4261 tactacccaa atatctggat agcatatgct atcctaatct atatctgggt agcatatgct
     4321 atcctaatct atatctgggt agcataggct atcctaatct atatctgggt agcatatgct
     4381 atcctaatct atatctgggt agtatatgct atcctaattt atatctgggt agcataggct
     4441 atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt agtatatgct
     4501 atcctaatct gtatccgggt agcatatgct atcctcatga taagctgtca aacatgagaa
     4561 ttttcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     4621 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     4681 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     4741 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     4801 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     4861 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     4921 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     4981 aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt
     5041 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     5101 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5161 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     5221 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5281 ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa
     5341 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     5401 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     5461 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     5521 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     5581 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     5641 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5701 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5761 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5821 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     5881 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     5941 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6001 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6061 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6121 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6181 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6241 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6301 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6361 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6421 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     6481 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     6541 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     6601 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     6661 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     6721 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     6781 aacagctatg accatgatta cgccaagctc tagctagagg tcgaccaatt ctcatgtttg
     6841 acagcttatc atcgcagatc cgggcaacgt tgttgccatt gctgcaggcg cagaactggt
     6901 aggtatggaa gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt
     6961 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata
     7021 tgtacattta tattggctca tgtccaatat gaccgccatg tt
//
