LOCUS       sequence-274887-        7755 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 7755)
  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 7755)
  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: D58_Ld_280280
FEATURES             Location/Qualifiers
     source          1..7755
                     /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             2533..2550
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2602..2657
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2656..2675)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4893..4911)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4979..5083
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5084..5152,5153..5944)
                     /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(5302..5321)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6115..6703
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6604..6623
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6857..6874
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6991..7012
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7027..7032,7033..7050,7051..7057)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7065..7081
                     /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     7070..7092
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7089..7105
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7089..7105
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7376..7755
                     /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 gccctggatg agcagcacag ctactacgag
      841 ggcaactacg gcaccaccgg cctggtgttc ggcagccccg atagcagccg gaccagctgg
      901 gactacacca acctgaagga ctggcccacc ctgtgcctga ccggcagaag gcagagcccc
      961 atcagcttcg ccaacgtgaa ccccgacgag gtggtcacca acgccctgct gcagcggctg
     1021 cagttcagct ctaagtgcgt gttcccccgg gaagagacac agatgcggat catcaacgag
     1081 ggcatcgtgt ccaccgtgtc cttcgaggaa ctgggccggt ccctggacga catgagcgag
     1141 tgcaccgtga tggaccccct gaacagaagc ctgacctacc acttcatggg cctgcacttc
     1201 cacgccatgc ccgtgcacca gctgagaacc ctgaggcccg acgccgagat gcacatgagc
     1261 ttcaccacca accacgtgga acagaaaatg cggaacgtgc tgatcgtggc cgtgatgctg
     1321 aaggccagcg ccatggtcaa cagcaccagc gccagagccc tgcagcacat cctggtggac
     1381 ggcagcctgc ccaagcggaa cgccatgacc acctgttttc tgaccgagag cctgagcctg
     1441 ttcagcctga tccccgccag agagagctac ctgctgtacg acggcagcca gacccacccc
     1501 ccctgcaccg agaacgtccg ctgggtcgtg atgaccagcc ccatcctgat cagccccgtg
     1561 gccctgggca gactgagaga tgccatggat gccctgctgc ccaacgactt ccacagattc
     1621 ggcaacgcca gaccccccca ggccctgaac ggccggctga tcttcagatt cgacgacaga
     1681 agcgtgcccg acggcggcaa ccggggcgag ggcaatttca aggacgcctg gtcccacaag
     1741 gacgagggct gggccaacag atccctgggc gtcagagccg gcagagtgct ggaaggcagc
     1801 ctggtggaag tggacggcga ctataagtac gacagcgccg tggtggtgtc ccctgtgtcc
     1861 cccaatcccg tgtccgccag ccggaatggc accacccacg ccagcagccc ccctatcacc
     1921 accagccccg gcagatacaa cggcagcagc gaccaggcca tggccaagcc tggaacaggc
     1981 gccctgggcc ctgcctacat ggccaacgcc agcagaaccg gccctgagac acacagccac
     2041 gtgaacgaga gcgacgccga ctacagcaac gccaccgcca atgccagctc cgcccccaga
     2101 accagcgtgg tgtctagcgc cagcgccagc agcgccgacg agagcatgcc ttctggcctg
     2161 caccacggca gcgagagcag cgcccctacc ccttctgacg agagcgcccc cagatccacc
     2221 accaccagca caaccaccac aactaccacc accaaagagc ccagcggcac caagggcaag
     2281 aagagatacg gcgacggcgc cagcaagaac accagcgaag ccgacgccgg cgtggtggac
     2341 agcgtgaagt ccatcgtgac ccgggccttc accagcgtga aggccttctc catcggcgcc
     2401 ttccagagca gcgtggccta cgccaaggcc aaccctggcg cgcccggcag cctgcaccac
     2461 atcctggacg cccagaagat gctgtggaac cacagggaca ggaacctgcc tcctctggcc
     2521 cctctgggcc cccaccacca tcaccaccat tgagtctaga cacgtgatta attaaggatc
     2581 ccccgacctc gacctctggc taataaagga aatttatttt cattgcaata gtgtgttgga
     2641 attttttgtg tctctcactc ggaaggacat atgggagggc aaatcatttg gtcgagatcc
     2701 ctcggagatc tctagctaga ggatcgatcc ccgccccgga cgaactaaac ctgactacga
     2761 catctctgcc ccttcttcgc ggggcagtgc atgtaatccc ttcagttggt tggtacaact
     2821 tgccaactgg gccctgttcc acatgtgaca cgggggggga ccaaacacaa aggggttctc
     2881 tgactgtagt tgacatcctt ataaatggat gtgcacattt gccaacactg agtggctttc
     2941 atcctggagc agactttgca gtctgtggac tgcaacacaa cattgccttt atgtgtaact
     3001 cttggctgaa gctcttacac caatgctggg ggacatgtac ctcccagggg cccaggaaga
     3061 ctacgggagg ctacaccaac gtcaatcaga ggggcctgtg tagctaccga taagcggacc
     3121 ctcaagaggg cattagcaat agtgtttata aggccccctt gttaacccta aacgggtagc
     3181 atatgcttcc cgggtagtag tatatactat ccagactaac cctaattcaa tagcatatgt
     3241 tacccaacgg gaagcatatg ctatcgaatt agggttagta aaagggtcct aaggaacagc
     3301 gatatctccc accccatgag ctgtcacggt tttatttaca tggggtcagg attccacgag
     3361 ggtagtgaac cattttagtc acaagggcag tggctgaaga tcaaggagcg ggcagtgaac
     3421 tctcctgaat cttcgcctgc ttcttcattc tccttcgttt agctaataga ataactgctg
     3481 agttgtgaac agtaaggtgt atgtgaggtg ctcgaaaaca aggtttcagg tgacgccccc
     3541 agaataaaat ttggacgggg ggttcagtgg tggcattgtg ctatgacacc aatataaccc
     3601 tcacaaaccc cttgggcaat aaatactagt gtaggaatga aacattctga atatctttaa
     3661 caatagaaat ccatggggtg gggacaagcc gtaaagactg gatgtccatc tcacacgaat
     3721 ttatggctat gggcaacaca taatcctagt gcaatatgat actggggtta ttaagatgtg
     3781 tcccaggcag ggaccaagac aggtgaacca tgttgttaca ctctatttgt aacaagggga
     3841 aagagagtgg acgccgacag cagcggactc cactggttgt ctctaacacc cccgaaaatt
     3901 aaacggggct ccacgccaat ggggcccata aacaaagaca agtggccact cttttttttg
     3961 aaattgtgga gtgggggcac gcgtcagccc ccacacgccg ccctgcggtt ttggactgta
     4021 aaataagggt gtaataactt ggctgattgt aaccccgcta accactgcgg tcaaaccact
     4081 tgcccacaaa accactaatg gcaccccggg gaatacctgc ataagtaggt gggcgggcca
     4141 agataggggc gcgattgctg cgatctggag gacaaattac acacacttgc gcctgagcgc
     4201 caagcacagg gttgttggtc ctcatattca cgaggtcgct gagagcacgg tgggctaatg
     4261 ttgccatggg tagcatatac tacccaaata tctggatagc atatgctatc ctaatctata
     4321 tctgggtagc ataggctatc ctaatctata tctgggtagc atatgctatc ctaatctata
     4381 tctgggtagt atatgctatc ctaatttata tctgggtagc ataggctatc ctaatctata
     4441 tctgggtagc atatgctatc ctaatctata tctgggtagt atatgctatc ctaatctgta
     4501 tccgggtagc atatgctatc ctaatagaga ttagggtagt atatgctatc ctaatttata
     4561 tctgggtagc atatactacc caaatatctg gatagcatat gctatcctaa tctatatctg
     4621 ggtagcatat gctatcctaa tctatatctg ggtagcatag gctatcctaa tctatatctg
     4681 ggtagcatat gctatcctaa tctatatctg ggtagtatat gctatcctaa tttatatctg
     4741 ggtagcatag gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg
     4801 ggtagtatat gctatcctaa tctgtatccg ggtagcatat gctatcctca tgataagctg
     4861 tcaaacatga gaattttctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt
     4921 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     4981 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     5041 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     5101 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     5161 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     5221 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     5281 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
     5341 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     5401 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     5461 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     5521 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     5581 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac
     5641 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     5701 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     5761 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     5821 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     5881 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     5941 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     6001 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     6061 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     6121 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     6181 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     6241 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     6301 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     6361 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     6421 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     6481 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6541 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6601 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6661 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6721 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     6781 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     6841 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     6901 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     6961 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
     7021 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
     7081 atttcacaca ggaaacagct atgaccatga ttacgccaag ctctagctag aggtcgacca
     7141 attctcatgt ttgacagctt atcatcgcag atccgggcaa cgttgttgcc attgctgcag
     7201 gcgcagaact ggtaggtatg gaagatctat acattgaatc aatattggca attagccata
     7261 ttagtcattg gttatatagc ataaatcaat attggctatt ggccattgca tacgttgtat
     7321 ctatatcata atatgtacat ttatattggc tcatgtccaa tatgaccgcc atgttgacat
     7381 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
     7441 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     7501 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     7561 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     7621 tatcatatgc caagtccgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     7681 tatgcccagt acatgacctt acgggacttt cctacttggc agtacatcta cgtattagtc
     7741 atcgctatta ccatg
//
