LOCUS       sequence-274512-        8502 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 8502)
  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 8502)
  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: D44_Ld_180090
FEATURES             Location/Qualifiers
     source          1..8502
                     /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             3660..3677
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3729..3784
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3783..3802)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6020..6038)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6106..6210
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6211..6279,6280..7071)
                     /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(6429..6448)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7242..7830
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7731..7750
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7984..8001
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8118..8139
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8154..8159,8160..8177,8178..8184)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8192..8208
                     /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     8197..8219
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8216..8232
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8216..8232
                     /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 ccgtggtcac aacagaaggc ggcctggccc tggaagccat cggctacccc aacgacgtcg
     1261 tgcggctgac cgccaagccc ggcgacaagt ggacctacca gcccgacggc atcatcctgc
     1321 ccgacgccgt ggctctgcag gccaccaaag gcgcctccga cgcccagtgg tccatccccc
     1381 acggcaagtg catgctgacc atcagcatgg cccctcccca cgtgcgcttc gagttcagct
     1441 gtcacggcag cagcctggtg gaagtgaccg ccttcgccga tgccatggct gcccccgaag
     1501 tgaacttcac cttccctgcc gcccagacta tgtacggact ggccgagcac gccgccgacc
     1561 tgcctctgag aggcggcaac gtgtacgaga tgtacaacac cgacaccttc cagtacagcg
     1621 tgaacagcac cgaggccctg tacggcgtga tccccttcat catggcctac gcccccaagt
     1681 ctacctgcgg cgtgctgttc ctgaacccca gcgagacaaa cgtgggcgtg tccgccgata
     1741 gcgccgctcc cagctgccag tggaagcctg aagtgggcgc catcgacatc ttttttctgc
     1801 ccggacccac ccccaccaag gtccagcagc agcatgccgc cctgaccggc gccacagtga
     1861 tgccccctta cttcagcctg ggcctgcacc agtgccggtg gaactacctg aacaccaagg
     1921 actgcctgag cgtggacgag ggcttcgaca cccacaacat gccctacgac gtgctgtggc
     1981 tggacatcga gcacaccgat aagaagaagt actttacctg ggacccctac acctttcccg
     2041 accccaaggc cctgacagac gccctggcca gcaagggcag aaagctggtc accgtgcggg
     2101 accctcacgt gaagcgggat gagggctact acgtgcacaa cgaggcccag aaaggcggct
     2161 actatgtgaa ggacgccagc ggcgaggact acgtgggcaa gtgttggccc ggcagctcca
     2221 gctggcccga cttcttcaac acccggacca gagtgtggta cagccagttc ttccacgacg
     2281 acagataccc tggcggcagc cgggacatcc acacctgggt ggacatgaac gagcccagcg
     2341 tgttcggcgg cgagcggggc acaatgccta agaccgccgt gcacagcctg gacaacggcc
     2401 acaccgtgga acacagattc gtgcataacg cctacagctt ctatagcgtg caggcagtgc
     2461 acaagggcat gctcgaggct ggcggcccta acaccgcccc tgagcggccc tttatcctga
     2521 ccagaagctt cttcagcggc tcccagagat acgccgccat gtggaccggc gataacatgg
     2581 ccagatggga ccacctggaa aacagcatcc ccgagctgct gagcctgagc atctccaact
     2641 accctttctg cggctgcgat atcggcggat tcttcttcga ccccgaagag gaactgttcg
     2701 tgcggtggat gcaggccggg gtgttcgtcc ctttctaccg ggcccacagc catctggaca
     2761 ccaagcggag agagccctgg accttcagcg ccgaagccca gtctctcgtg cgcagcgccc
     2821 tcgccctgcg ctacgccatg ctgccttacc tgtacacaac cttctaccac gcccacaccg
     2881 agggcaacac catcatgcgg cccctgttct acgagttccc ccggcagagc gagctgagag
     2941 aggtgcagaa cacctacctg ttcggcccca gcatcctcgt ccggcccgtc gtgaagccta
     3001 gcgtgaccga agtcacagtg cccctgccca aagaggccct ctggtacaac tacttctccg
     3061 gcgagctggc cgtgggccag cacaccatgc ccgtggacaa ggacaccatc cccatgtttc
     3121 tgaggggcgg acacatcgtg cccatgaagc tgagactgcg gcggagcagc ttcgccgcca
     3181 gactggaccc tttcaccctg tttgtggccc tgaatgccca gggcaactcc tacggcgatc
     3241 tgtacatcga cgacggcacc acctacgact acaaaaaggg cgccttcgtg caccgggcct
     3301 tcagctacag caaccaggtg ctgcagaact ccgcctaccc taacagcccc ggcaccagcg
     3361 gcctgtacaa cgccgtgaat gccgtggaaa gagtggtcat ctacggctat gtgggaaagg
     3421 tgtccaaggc cgtgctgaat acccgggtgg acgaaaccga ggtggccacc cccctggaat
     3481 tcgagcagct gggccagaca gtggtcgtcc ggaagccttc tgtgctggtg gccgccgatt
     3541 ggatcatcag cttcgagaac gaaggcgcgc ccggcagcct gcaccacatc ctggacgccc
     3601 agaagatgct gtggaaccac agggacagga acctgcctcc tctggcccct ctgggccccc
     3661 accaccatca ccaccattga gtctagacac gtgattaatt aaggatcccc cgacctcgac
     3721 ctctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct
     3781 ctcactcgga aggacatatg ggagggcaaa tcatttggtc gagatccctc ggagatctct
     3841 agctagagga tcgatccccg ccccggacga actaaacctg actacgacat ctctgcccct
     3901 tcttcgcggg gcagtgcatg taatcccttc agttggttgg tacaacttgc caactgggcc
     3961 ctgttccaca tgtgacacgg ggggggacca aacacaaagg ggttctctga ctgtagttga
     4021 catccttata aatggatgtg cacatttgcc aacactgagt ggctttcatc ctggagcaga
     4081 ctttgcagtc tgtggactgc aacacaacat tgcctttatg tgtaactctt ggctgaagct
     4141 cttacaccaa tgctggggga catgtacctc ccaggggccc aggaagacta cgggaggcta
     4201 caccaacgtc aatcagaggg gcctgtgtag ctaccgataa gcggaccctc aagagggcat
     4261 tagcaatagt gtttataagg cccccttgtt aaccctaaac gggtagcata tgcttcccgg
     4321 gtagtagtat atactatcca gactaaccct aattcaatag catatgttac ccaacgggaa
     4381 gcatatgcta tcgaattagg gttagtaaaa gggtcctaag gaacagcgat atctcccacc
     4441 ccatgagctg tcacggtttt atttacatgg ggtcaggatt ccacgagggt agtgaaccat
     4501 tttagtcaca agggcagtgg ctgaagatca aggagcgggc agtgaactct cctgaatctt
     4561 cgcctgcttc ttcattctcc ttcgtttagc taatagaata actgctgagt tgtgaacagt
     4621 aaggtgtatg tgaggtgctc gaaaacaagg tttcaggtga cgcccccaga ataaaatttg
     4681 gacggggggt tcagtggtgg cattgtgcta tgacaccaat ataaccctca caaacccctt
     4741 gggcaataaa tactagtgta ggaatgaaac attctgaata tctttaacaa tagaaatcca
     4801 tggggtgggg acaagccgta aagactggat gtccatctca cacgaattta tggctatggg
     4861 caacacataa tcctagtgca atatgatact ggggttatta agatgtgtcc caggcaggga
     4921 ccaagacagg tgaaccatgt tgttacactc tatttgtaac aaggggaaag agagtggacg
     4981 ccgacagcag cggactccac tggttgtctc taacaccccc gaaaattaaa cggggctcca
     5041 cgccaatggg gcccataaac aaagacaagt ggccactctt ttttttgaaa ttgtggagtg
     5101 ggggcacgcg tcagccccca cacgccgccc tgcggttttg gactgtaaaa taagggtgta
     5161 ataacttggc tgattgtaac cccgctaacc actgcggtca aaccacttgc ccacaaaacc
     5221 actaatggca ccccggggaa tacctgcata agtaggtggg cgggccaaga taggggcgcg
     5281 attgctgcga tctggaggac aaattacaca cacttgcgcc tgagcgccaa gcacagggtt
     5341 gttggtcctc atattcacga ggtcgctgag agcacggtgg gctaatgttg ccatgggtag
     5401 catatactac ccaaatatct ggatagcata tgctatccta atctatatct gggtagcata
     5461 ggctatccta atctatatct gggtagcata tgctatccta atctatatct gggtagtata
     5521 tgctatccta atttatatct gggtagcata ggctatccta atctatatct gggtagcata
     5581 tgctatccta atctatatct gggtagtata tgctatccta atctgtatcc gggtagcata
     5641 tgctatccta atagagatta gggtagtata tgctatccta atttatatct gggtagcata
     5701 tactacccaa atatctggat agcatatgct atcctaatct atatctgggt agcatatgct
     5761 atcctaatct atatctgggt agcataggct atcctaatct atatctgggt agcatatgct
     5821 atcctaatct atatctgggt agtatatgct atcctaattt atatctgggt agcataggct
     5881 atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt agtatatgct
     5941 atcctaatct gtatccgggt agcatatgct atcctcatga taagctgtca aacatgagaa
     6001 ttttcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     6061 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     6121 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     6181 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     6241 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     6301 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     6361 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     6421 aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt
     6481 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     6541 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     6601 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     6661 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     6721 ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa
     6781 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     6841 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     6901 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     6961 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     7021 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     7081 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     7141 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     7201 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     7261 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     7321 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     7381 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     7441 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     7501 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     7561 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     7621 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     7681 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     7741 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     7801 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     7861 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     7921 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     7981 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     8041 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     8101 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     8161 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     8221 aacagctatg accatgatta cgccaagctc tagctagagg tcgaccaatt ctcatgtttg
     8281 acagcttatc atcgcagatc cgggcaacgt tgttgccatt gctgcaggcg cagaactggt
     8341 aggtatggaa gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt
     8401 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata
     8461 tgtacattta tattggctca tgtccaatat gaccgccatg tt
//
