LOCUS       sequence-275081-        8049 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 8049)
  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 8049)
  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: D32_Ld_354780
FEATURES             Location/Qualifiers
     source          1..8049
                     /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             3010..3027
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3079..3134
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3133..3152)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     CDS             join(5378..5446,5447..6238)
                     /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(5596..5615)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6409..6997
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6898..6917
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7151..7168
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7285..7306
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7321..7326,7327..7344,7345..7351)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7359..7375
                     /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     7364..7386
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7383..7399
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7383..7399
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7670..8049
                     /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 gccacccacg tgatcggagt ggatctgggc
      841 agcgagtaca tcaaggtggc cggaccccac ggcgacaagg gcgtggacat cgtgctgaac
      901 gagcagagca gaagaaagac cgacaacttc atcggcttcc ggcggagcga cctgtacatc
      961 ggcgacaccg ccaagagcct ggccgccaga ttccctctgt gtaccgccag cgccgtgaac
     1021 cagctcgtgc ggatccggaa ggacagcagc ctgctgccat tcttccggga cctgcagtac
     1081 gagtaccacg tgtccttcaa caaccacggc agcgccaccg tgtccatctg cgataccaag
     1141 gaccccttca ccgccgagga actgtacagc atggtgctga gctactgcaa gaccacagcc
     1201 gtgaaggacg atgtggtgga ccccaagggg gtggtggtca ccatcccctt cagaaccagc
     1261 cctgccgagc ggagagccat cctggacgcc gctagactga gcggcctgag cgtgctgggc
     1321 ctgatgcaca gcaccaccgc cgctgccttc tactacggcg tgcggcaccg gggcttcggc
     1381 aacaacaccc tgagagtggt ggtgttcgac ctgggctcca cccacaccga agtgggcgtg
     1441 tacgagttcc tgcctgccgc ccagaaggcc cccctgagca gcgcctttgg cacactgaga
     1501 accctgggcg tggtggaaga tcggagcctg ggcggcagag ccttcgatct gtgcgtggcc
     1561 agagtgatcg aggccgaggc cagagccaag ctgagcatcg gccctgtgct gggaggcacc
     1621 accagcgccc agctgaagtc ccagttctcc ctgctgagag ccgccaacaa agtgcgggaa
     1681 accctgtccg tgaacagcaa caccccctac accgtggaag ggatcgcccc cgaccgggac
     1741 taccacagca gcatgaccag ggccaccttc gaagccgagt gcgccagcct gttccagaga
     1801 gtgaaggatc tggccacctc cgtgaccaag gtcgtgaaca tcagcttcaa ggacctgaac
     1861 agcttcgaga tgatgggcgg cctgtccaga acccccaaga tcattgccga cctgagcgag
     1921 gccatcggcc gggacgtgga ccggaccatg aacatggacg aggctgccgc cattggagcc
     1981 ggctactacg ccgccaaact gtcccccctg taccgggcca agtccctgaa gctggacgag
     2041 acaatcccct acggggtgga cttcgaggtg gaaccccctc tgtccaaaga gaagcccgcc
     2101 gtcagacggc ccctgttcgg cgccgatggc ttcctgctgg gcgacagcgt gtccctgacc
     2161 ttcaaccgga ccgaggactt cagcctgaac ttctacatcg atgacgatgc cgccaggccc
     2221 ttcgtgcgcg tgaccgtgac aggcgtgaag caggccctga ccgacatgaa ggccctgtcc
     2281 ccagccgtgg aacatgccaa caacagccac atggtccgac tgcagattgt gctgaatgag
     2341 acaggcctgg tgcaggtcga gcacgccgag gtggtcgtca gatacgccga agaagtgacc
     2401 cagaaagtca ccgagaacgt gaccgataag gccagcggcg agcggaagtc cgtggaaaag
     2461 tctgtgaaag tgatcaagat gcggatcaga gccgctgacg tgacagccac cctggcctgg
     2521 aagaacccca ccggcctgac cagcgaggaa atggaaacca gccagacaaa gctgaaaacc
     2581 atctggcagg ccgaacacgt gaaacacctg agggccactg ccaagaacaa cctggaagcc
     2641 tacatcttct gggccaagaa tgagggcgtg tccgacaacg ccgccctgac agctgctgct
     2701 ggcgctgccg ctgtgcagcg cgtgatcgat gaagccaccg ccatgcagga atggctggaa
     2761 gagggcgagg gcgcctccga taattgcgcc gccagccagt atgagagcag actggaaaac
     2821 ctgcggggca tggtggccga gctgaccaag cagcccgaag aggccaccaa gacagtgacc
     2881 accgactccg ccgacgacaa cgagggcgac ctgggcgcgc ccggcagcct gcaccacatc
     2941 ctggacgccc agaagatgct gtggaaccac agggacagga acctgcctcc tctggcccct
     3001 ctgggccccc accaccatca ccaccattga gtctagacac gtgattaatt aaggatcccc
     3061 cgacctcgac ctctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt
     3121 ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttggtc gagatccctc
     3181 ggagatctct agctagagga tcgatccccg ccccggacga actaaacctg actacgacat
     3241 ctctgcccct tcttcgcggg gcagtgcatg taatcccttc agttggttgg tacaacttgc
     3301 caactgggcc ctgttccaca tgtgacacgg ggggggacca aacacaaagg ggttctctga
     3361 ctgtagttga catccttata aatggatgtg cacatttgcc aacactgagt ggctttcatc
     3421 ctggagcaga ctttgcagtc tgtggactgc aacacaacat tgcctttatg tgtaactctt
     3481 ggctgaagct cttacaccaa tgctggggga catgtacctc ccaggggccc aggaagacta
     3541 cgggaggcta caccaacgtc aatcagaggg gcctgtgtag ctaccgataa gcggaccctc
     3601 aagagggcat tagcaatagt gtttataagg cccccttgtt aaccctaaac gggtagcata
     3661 tgcttcccgg gtagtagtat atactatcca gactaaccct aattcaatag catatgttac
     3721 ccaacgggaa gcatatgcta tcgaattagg gttagtaaaa gggtcctaag gaacagcgat
     3781 atctcccacc ccatgagctg tcacggtttt atttacatgg ggtcaggatt ccacgagggt
     3841 agtgaaccat tttagtcaca agggcagtgg ctgaagatca aggagcgggc agtgaactct
     3901 cctgaatctt cgcctgcttc ttcattctcc ttcgtttagc taatagaata actgctgagt
     3961 tgtgaacagt aaggtgtatg tgaggtgctc gaaaacaagg tttcaggtga cgcccccaga
     4021 ataaaatttg gacggggggt tcagtggtgg cattgtgcta tgacaccaat ataaccctca
     4081 caaacccctt gggcaataaa tactagtgta ggaatgaaac attctgaata tctttaacaa
     4141 tagaaatcca tggggtgggg acaagccgta aagactggat gtccatctca cacgaattta
     4201 tggctatggg caacacataa tcctagtgca atatgatact ggggttatta agatgtgtcc
     4261 caggcaggga ccaagacagg tgaaccatgt tgttacactc tatttgtaac aaggggaaag
     4321 agagtggacg ccgacagcag cggactccac tggttgtctc taacaccccc gaaaattaaa
     4381 cggggctcca cgccaatggg gcccataaac aaagacaagt ggccactctt ttttttgaaa
     4441 ttgtggagtg ggggcacgcg tcagccccca cacgccgccc tgcggttttg gactgtaaaa
     4501 taagggtgta ataacttggc tgattgtaac cccgctaacc actgcggtca aaccacttgc
     4561 ccacaaaacc actaatggca ccccggggaa tacctgcata agtaggtggg cgggccaaga
     4621 taggggcgcg attgctgcga tctggaggac aaattacaca cacttgcgcc tgagcgccaa
     4681 gcacagggtt gttggtcctc atattcacga ggtcgctgag agcacggtgg gctaatgttg
     4741 ccatgggtag catatactac ccaaatatct ggatagcata tgctatccta atctatatct
     4801 gggtagcata ggctatccta atctatatct gggtagcata tgctatccta atctatatct
     4861 gggtagtata tgctatccta atttatatct gggtagcata ggctatccta atctatatct
     4921 gggtagcata tgctatccta atctatatct gggtagtata tgctatccta atctgtatcc
     4981 gggtagcata tgctatccta atagagatta gggtagtata tgctatccta atttatatct
     5041 gggtagcata tactacccaa atatctggat agcatatgct atcctaatct atatctgggt
     5101 agcatatgct atcctaatct atatctgggt agcataggct atcctaatct atatctgggt
     5161 agcatatgct atcctaatct atatctgggt agtatatgct atcctaattt atatctgggt
     5221 agcataggct atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt
     5281 agtatatgct atcctaatct gtatccgggt agcatatgct atcctcatga gacaataacc
     5341 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     5401 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     5461 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     5521 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     5581 cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca
     5641 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     5701 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     5761 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     5821 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     5881 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt
     5941 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     6001 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     6061 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     6121 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     6181 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     6241 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     6301 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     6361 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     6421 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     6481 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     6541 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     6601 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     6661 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     6721 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     6781 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6841 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6901 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6961 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     7021 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     7081 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     7141 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     7201 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     7261 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
     7321 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     7381 cacaggaaac agctatgacc atgattacgc caagctctag ctagaggtcg accaattctc
     7441 atgtttgaca gcttatcatc gcagatccgg gcaacgttgt tgccattgct gcaggcgcag
     7501 aactggtagg tatggaagat ctatacattg aatcaatatt ggcaattagc catattagtc
     7561 attggttata tagcataaat caatattggc tattggccat tgcatacgtt gtatctatat
     7621 cataatatgt acatttatat tggctcatgt ccaatatgac cgccatgttg acattgatta
     7681 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
     7741 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
     7801 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
     7861 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
     7921 atgccaagtc cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
     7981 cagtacatga ccttacggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     8041 attaccatg
//
