LOCUS       sequence-274675-        6855 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 6855)
  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 6855)
  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: D89_Ld_343450
FEATURES             Location/Qualifiers
     source          1..6855
                     /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     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             2013..2030
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2082..2137
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2136..2155)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4373..4391)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4459..4563
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4564..4632,4633..5424)
                     /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(4782..4801)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5595..6183
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6084..6103
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6337..6354
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6471..6492
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6507..6512,6513..6530,6531..6537)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6545..6561
                     /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     6550..6572
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6569..6585
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6569..6585
                     /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 gccggaatta
     1201 caggttattc tactggtacg atcatttctc cccaggctaa cgcatatcgg agtcggcgca
     1261 cactgagccc aatggattat tacaaactgc ctgtatgtca gccgtctgac gaggtcatga
     1321 aggcaaggcg tgagcacccc ctcattgggg atattctgac tgggaaccgt ctggtgccca
     1381 caatgtttga attcagggtc ggcgaggatg taaagtgtgc caccctctgc gatgcctctt
     1441 tcaccgtgaa agccgtgcgg cgcgcgaact acatgattaa taatgactac tatgtgcgga
     1501 tgttcctgga taacaaaccg ctcgtctctg catctcccca cgagggcagc aacgcgtacc
     1561 tgcttggtta tccacttggc gcccagaatg acgttgagaa aacccaggtg aaaaccaata
     1621 tcattcacaa tcaccttgac ttcacaataa gaattaagaa tcgcgcgatt tctcagttta
     1681 ccggcgaaga ggtggtgggt tttaaggtcg tggctaggtc actcgctgaa gtgggtacct
     1741 gcacggctac cgctttccag cactcttcca ggccctacat ccttccaagc taccgcgacg
     1801 ggaaggacct gaaagtgccc tttacctaca gcgtgacctg ggagcgcagc aacgaggagt
     1861 atccaatcga gcaccgcatc ggagaggaca cgcagaggcg cggccataag atcgccggcg
     1921 cgcccggcag cctgcaccac atcctggacg cccagaagat gctgtggaac cacagggaca
     1981 ggaacctgcc tcctctggcc cctctgggcc cccaccacca tcaccaccat tgagtctaga
     2041 cacgtgatta attaaggatc ccccgacctc gacctctggc taataaagga aatttatttt
     2101 cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc
     2161 aaatcatttg gtcgagatcc ctcggagatc tctagctaga ggatcgatcc ccgccccgga
     2221 cgaactaaac ctgactacga catctctgcc ccttcttcgc ggggcagtgc atgtaatccc
     2281 ttcagttggt tggtacaact tgccaactgg gccctgttcc acatgtgaca cgggggggga
     2341 ccaaacacaa aggggttctc tgactgtagt tgacatcctt ataaatggat gtgcacattt
     2401 gccaacactg agtggctttc atcctggagc agactttgca gtctgtggac tgcaacacaa
     2461 cattgccttt atgtgtaact cttggctgaa gctcttacac caatgctggg ggacatgtac
     2521 ctcccagggg cccaggaaga ctacgggagg ctacaccaac gtcaatcaga ggggcctgtg
     2581 tagctaccga taagcggacc ctcaagaggg cattagcaat agtgtttata aggccccctt
     2641 gttaacccta aacgggtagc atatgcttcc cgggtagtag tatatactat ccagactaac
     2701 cctaattcaa tagcatatgt tacccaacgg gaagcatatg ctatcgaatt agggttagta
     2761 aaagggtcct aaggaacagc gatatctccc accccatgag ctgtcacggt tttatttaca
     2821 tggggtcagg attccacgag ggtagtgaac cattttagtc acaagggcag tggctgaaga
     2881 tcaaggagcg ggcagtgaac tctcctgaat cttcgcctgc ttcttcattc tccttcgttt
     2941 agctaataga ataactgctg agttgtgaac agtaaggtgt atgtgaggtg ctcgaaaaca
     3001 aggtttcagg tgacgccccc agaataaaat ttggacgggg ggttcagtgg tggcattgtg
     3061 ctatgacacc aatataaccc tcacaaaccc cttgggcaat aaatactagt gtaggaatga
     3121 aacattctga atatctttaa caatagaaat ccatggggtg gggacaagcc gtaaagactg
     3181 gatgtccatc tcacacgaat ttatggctat gggcaacaca taatcctagt gcaatatgat
     3241 actggggtta ttaagatgtg tcccaggcag ggaccaagac aggtgaacca tgttgttaca
     3301 ctctatttgt aacaagggga aagagagtgg acgccgacag cagcggactc cactggttgt
     3361 ctctaacacc cccgaaaatt aaacggggct ccacgccaat ggggcccata aacaaagaca
     3421 agtggccact cttttttttg aaattgtgga gtgggggcac gcgtcagccc ccacacgccg
     3481 ccctgcggtt ttggactgta aaataagggt gtaataactt ggctgattgt aaccccgcta
     3541 accactgcgg tcaaaccact tgcccacaaa accactaatg gcaccccggg gaatacctgc
     3601 ataagtaggt gggcgggcca agataggggc gcgattgctg cgatctggag gacaaattac
     3661 acacacttgc gcctgagcgc caagcacagg gttgttggtc ctcatattca cgaggtcgct
     3721 gagagcacgg tgggctaatg ttgccatggg tagcatatac tacccaaata tctggatagc
     3781 atatgctatc ctaatctata tctgggtagc ataggctatc ctaatctata tctgggtagc
     3841 atatgctatc ctaatctata tctgggtagt atatgctatc ctaatttata tctgggtagc
     3901 ataggctatc ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt
     3961 atatgctatc ctaatctgta tccgggtagc atatgctatc ctaatagaga ttagggtagt
     4021 atatgctatc ctaatttata tctgggtagc atatactacc caaatatctg gatagcatat
     4081 gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagcatag
     4141 gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagtatat
     4201 gctatcctaa tttatatctg ggtagcatag gctatcctaa tctatatctg ggtagcatat
     4261 gctatcctaa tctatatctg ggtagtatat gctatcctaa tctgtatccg ggtagcatat
     4321 gctatcctca tgataagctg tcaaacatga gaattttctt gaagacgaaa gggcctcgtg
     4381 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     4441 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     4501 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     4561 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     4621 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     4681 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     4741 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     4801 tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     4861 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     4921 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     4981 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     5041 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     5101 atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     5161 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     5221 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     5281 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     5341 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     5401 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     5461 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     5521 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5581 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     5641 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     5701 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     5761 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     5821 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     5881 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     5941 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6001 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6061 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6121 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6181 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     6241 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     6301 gctgataccg ctcgccacag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     6361 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     6421 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     6481 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     6541 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     6601 ctctagctag aggtcgacca attctcatgt ttgacagctt atcatcgcag atccgggcaa
     6661 cgttgttgcc attgctgcag gcgcagaact ggtaggtatg gaagatctat acattgaatc
     6721 aatattggca attagccata ttagtcattg gttatatagc ataaatcaat attggctatt
     6781 ggccattgca tacgttgtat ctatatcata atatgtacat ttatattggc tcatgtccaa
     6841 tatgaccgcc atgtt
//
