LOCUS       sequence-274515-        7704 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 7704)
  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 7704)
  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: D5_Ld_280600
FEATURES             Location/Qualifiers
     source          1..7704
                     /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"
     regulatory      2106..2115
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2862..2879
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2931..2986
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2985..3004)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5222..5240)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5308..5412
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5413..5481,5482..6273)
                     /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(5631..5650)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6444..7032
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6933..6952
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7186..7203
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7320..7341
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7356..7361,7362..7379,7380..7386)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7394..7410
                     /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     7399..7421
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7418..7434
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7418..7434
                     /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 gccaccgctc
     1201 agggcgatcc tgagagagcc gacagcgagg aacccagatg cggcttcgac gagctggaag
     1261 cccggatgat cggcacccgg gtgtccgtga tcagccgggt ggaacctcct accggcgaac
     1321 tggccgtggc cgctgctgct acaggcgcct ggcagcccat ccggatcgcc gtgttcaccg
     1381 aggacatcag caacagcagc cagcactgca ccgccagcgg ccagagcaga cccaccttca
     1441 gaggcggcag agtgacctgc tctgccgccg acgtgctgac ccgcgagaag aaacgggtgc
     1501 tgctggaact gctgatcccc agcgccgtgc agctgcatca ggaacggctg aacgtccagc
     1561 gcgagaacgg caacatcgtg gtgtccccat tcatcaagaa gcacagcatc tgcggccagt
     1621 tcagcatccc cgaggaacac atgaagaccg gcgtgcccga cgccgacttc gtgctgtaca
     1681 tgagcgccgc tcccaccagc ggctctgtga tcgcctgggc cgtgaagtgc cagagcttcg
     1741 acaacggccg gtccagcgtg ggcgtggcca ccatcagccc caagtacatc accgccgagc
     1801 ccaagaccgt gcgggtggtg gcccatgaag tgctgcacgc cctgggcttc accagaagcg
     1861 tgttcaagca gcagaacatg ctggtcatgg ccagcttccg gggcaagagc cccagccccg
     1921 tgatcagatc cgccaacgtg gtggctcagg cccagctgca ctacggctgc aagacccagg
     1981 ccagcatgga actggaagat gagggcggca agggcaccgt gtccagccac tggaagcggc
     2041 ggagcgccaa ggatgagctg atggccggct tcagcggcgt gggagtgtac agcgccctga
     2101 caatcgccgc catggaagat accggctact accagggcaa ctacgccaag gccgagccca
     2161 tggcctacgg ccacgaagtg ggctgcaagc tgagcagcga gagatgcgtg atcaagagca
     2221 ccagccagat ccccgccatg ttctgcgacg cccccgatgc cccttggagc tgcaccagcg
     2281 atcggagagg catcggccgg tgcatcctga ccagctacaa cagcaacctg cccacctact
     2341 tccagtactt cggcgacccc agactgggcg gacccgaccc cctgatggac ttctgccctt
     2401 tcgtgcgggc tgccgacgac accatgtgcg ccgccaagac caacgccctg aagggcagcg
     2461 tgtacggcgt gatgagcaga tgcgtggaca cccctgccgg cttctccatc gacgactctg
     2521 ccgtgcagca gcacggaatc tgcgccgagg tgcagtgcgg cagcagcgcc tacggcgtga
     2581 agatcaacgg cgccagcgcc ttcagagact gcccccctgg cagcacctac aacctgagca
     2641 ccctgagccc cagcttcagc aagggccacc tcgtgtgccc cagctacgag agcgtgtgcg
     2701 ccatcaacat caacgccagc ctgtacgaag agtacagccg gctgctgacc gaccacagcg
     2761 tgacaggcgc gcccggcagc ctgcaccaca tcctggacgc ccagaagatg ctgtggaacc
     2821 acagggacag gaacctgcct cctctggccc ctctgggccc ccaccaccat caccaccatt
     2881 gagtctagac acgtgattaa ttaaggatcc cccgacctcg acctctggct aataaaggaa
     2941 atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata
     3001 tgggagggca aatcatttgg tcgagatccc tcggagatct ctagctagag gatcgatccc
     3061 cgccccggac gaactaaacc tgactacgac atctctgccc cttcttcgcg gggcagtgca
     3121 tgtaatccct tcagttggtt ggtacaactt gccaactggg ccctgttcca catgtgacac
     3181 ggggggggac caaacacaaa ggggttctct gactgtagtt gacatcctta taaatggatg
     3241 tgcacatttg ccaacactga gtggctttca tcctggagca gactttgcag tctgtggact
     3301 gcaacacaac attgccttta tgtgtaactc ttggctgaag ctcttacacc aatgctgggg
     3361 gacatgtacc tcccaggggc ccaggaagac tacgggaggc tacaccaacg tcaatcagag
     3421 gggcctgtgt agctaccgat aagcggaccc tcaagagggc attagcaata gtgtttataa
     3481 ggcccccttg ttaaccctaa acgggtagca tatgcttccc gggtagtagt atatactatc
     3541 cagactaacc ctaattcaat agcatatgtt acccaacggg aagcatatgc tatcgaatta
     3601 gggttagtaa aagggtccta aggaacagcg atatctccca ccccatgagc tgtcacggtt
     3661 ttatttacat ggggtcagga ttccacgagg gtagtgaacc attttagtca caagggcagt
     3721 ggctgaagat caaggagcgg gcagtgaact ctcctgaatc ttcgcctgct tcttcattct
     3781 ccttcgttta gctaatagaa taactgctga gttgtgaaca gtaaggtgta tgtgaggtgc
     3841 tcgaaaacaa ggtttcaggt gacgccccca gaataaaatt tggacggggg gttcagtggt
     3901 ggcattgtgc tatgacacca atataaccct cacaaacccc ttgggcaata aatactagtg
     3961 taggaatgaa acattctgaa tatctttaac aatagaaatc catggggtgg ggacaagccg
     4021 taaagactgg atgtccatct cacacgaatt tatggctatg ggcaacacat aatcctagtg
     4081 caatatgata ctggggttat taagatgtgt cccaggcagg gaccaagaca ggtgaaccat
     4141 gttgttacac tctatttgta acaaggggaa agagagtgga cgccgacagc agcggactcc
     4201 actggttgtc tctaacaccc ccgaaaatta aacggggctc cacgccaatg gggcccataa
     4261 acaaagacaa gtggccactc ttttttttga aattgtggag tgggggcacg cgtcagcccc
     4321 cacacgccgc cctgcggttt tggactgtaa aataagggtg taataacttg gctgattgta
     4381 accccgctaa ccactgcggt caaaccactt gcccacaaaa ccactaatgg caccccgggg
     4441 aatacctgca taagtaggtg ggcgggccaa gataggggcg cgattgctgc gatctggagg
     4501 acaaattaca cacacttgcg cctgagcgcc aagcacaggg ttgttggtcc tcatattcac
     4561 gaggtcgctg agagcacggt gggctaatgt tgccatgggt agcatatact acccaaatat
     4621 ctggatagca tatgctatcc taatctatat ctgggtagca taggctatcc taatctatat
     4681 ctgggtagca tatgctatcc taatctatat ctgggtagta tatgctatcc taatttatat
     4741 ctgggtagca taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat
     4801 ctgggtagta tatgctatcc taatctgtat ccgggtagca tatgctatcc taatagagat
     4861 tagggtagta tatgctatcc taatttatat ctgggtagca tatactaccc aaatatctgg
     4921 atagcatatg ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg
     4981 gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg
     5041 gtagtatatg ctatcctaat ttatatctgg gtagcatagg ctatcctaat ctatatctgg
     5101 gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat ctgtatccgg
     5161 gtagcatatg ctatcctcat gataagctgt caaacatgag aattttcttg aagacgaaag
     5221 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     5281 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     5341 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     5401 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     5461 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     5521 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     5581 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     5641 gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     5701 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     5761 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     5821 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     5881 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     5941 gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     6001 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     6061 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     6121 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     6181 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     6241 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     6301 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     6361 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6421 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6481 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     6541 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     6601 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     6661 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6721 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     6781 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     6841 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6901 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6961 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     7021 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     7081 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     7141 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     7201 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     7261 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     7321 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     7381 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     7441 tacgccaagc tctagctaga ggtcgaccaa ttctcatgtt tgacagctta tcatcgcaga
     7501 tccgggcaac gttgttgcca ttgctgcagg cgcagaactg gtaggtatgg aagatctata
     7561 cattgaatca atattggcaa ttagccatat tagtcattgg ttatatagca taaatcaata
     7621 ttggctattg gccattgcat acgttgtatc tatatcataa tatgtacatt tatattggct
     7681 catgtccaat atgaccgcca tgtt
//
