LOCUS       sequence-274897-        7854 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 7854)
  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 7854)
  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: D40_Ld_310530
FEATURES             Location/Qualifiers
     source          1..7854
                     /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             3012..3029
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3081..3136
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3135..3154)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5372..5390)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5458..5562
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5563..5631,5632..6423)
                     /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(5781..5800)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6594..7182
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7083..7102
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7336..7353
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7470..7491
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7506..7511,7512..7529,7530..7536)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7544..7560
                     /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     7549..7571
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7568..7584
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7568..7584
                     /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 gccaccctgc
     1201 ctcagcagga agcccctccc gtgatggaac agcctaccgc cgagcctgcc gaggccgatg
     1261 gacctgctag aggacctgcc cctccacccc catctggcag accagcccct aatgcccctg
     1321 gcgcctctcc ccctgccatc ggcgattctg gcgaaacccc tgcccagggc gagacactgg
     1381 accccatctt cgaggaaatc accagcctgt tcgccagccg gaaccccaag cacgccctgc
     1441 agggactgag cgacggcctg agaaatgccg ccacaggcgt gggactgggc ctggctggcg
     1501 tggtggctgg aacctacggc ggagccaaag agggcggcct gggcggcatg gccaaaggct
     1561 ttggagccgg cgtggccggc tttgtgggcc tgacaggata tggcgcctac gcaggcgtgc
     1621 ggcagatcgt gcggggcgtg ggcaatacca gatccgccgt gtccgaggtg tccaagggcg
     1681 aggcctactg ggacagcaac gcccaggctt gggtccgagt gaacctggcc tgcgacttcg
     1741 agcggctgcc ccctaccgat gaggatctgc tgggagaggc ccggaaggcc tacgagaagg
     1801 ccaagaagga cggcacactg cccacaaccc tgcccgccat gacccccaat gatagcgccg
     1861 agggcggaga agccagcgcc agctctgcta caacaggcgc tggcgccgct agcctgcccc
     1921 agggaaccga taccggcgag actaagcacg ctgccgccga acccgtgaac tactacgcct
     1981 tcctgggcgt ggaaagcacc gccaccagca gcgagatcag aaaggcctat acccgcaagg
     2041 ccctggaaat gcaccccgac aagaacccta acgaccccaa cgccaccatc aagttccagg
     2101 aactgaacaa gatctacaac gtgctgagcc acgaggacac ccgggccacc tacgaccagt
     2161 acggcaccgt ggaccctatg aacgtgcccg agatgaccgg caaccccatg aaggaactgc
     2221 tcggagccgc ctttctggaa gccctcgtgg gccccctgca cttctttctg gtgttcgagg
     2281 ggggcgtgct gttcacagcc gagatgcaga gagagctgca cgccagacgg cggctgcggg
     2341 tggccaaaaa cctgatcagc tggctggaca acgacgtgtc cggcttcgag agcgcccagc
     2401 tggccctgag ggatgccgtg tctacagccc tgggccccgt gttcgtgtcc tacgtggccg
     2461 aggaatacca cctggccagc agacagcagc tgcatggctc cagctggaag agagagatgg
     2521 acagctggta ctcctcctgg gccacatccg ccagcagcct gtggcattgg accaccatgg
     2581 gcgccagaac cgccaggcgg gccttcgtgg acaagaatct gggcgaagag gacatcctga
     2641 gagtgctggc cgtggccaac gagcgggacg tccgccagat tgtgctgcag gcctgccggc
     2701 tggtgctgtt tgacatgagc gtgaccccag cccagcggca gcagcgggcc atgagactgg
     2761 aagatctgtc tgtgatggct atgaaggaag tggccctcga ggtcgccagc agagagcgga
     2821 ttggagccag cgaagccaca gccgccgctg ccgagaaggg cgccacaaca aagacacccg
     2881 ccgacaccag ctccgtgacc gccccacctc tgtctggcgc gcccggcagc ctgcaccaca
     2941 tcctggacgc ccagaagatg ctgtggaacc acagggacag gaacctgcct cctctggccc
     3001 ctctgggccc ccaccaccat caccaccatt gagtctagac acgtgattaa ttaaggatcc
     3061 cccgacctcg acctctggct aataaaggaa atttattttc attgcaatag tgtgttggaa
     3121 ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttgg tcgagatccc
     3181 tcggagatct ctagctagag gatcgatccc cgccccggac gaactaaacc tgactacgac
     3241 atctctgccc cttcttcgcg gggcagtgca tgtaatccct tcagttggtt ggtacaactt
     3301 gccaactggg ccctgttcca catgtgacac ggggggggac caaacacaaa ggggttctct
     3361 gactgtagtt gacatcctta taaatggatg tgcacatttg ccaacactga gtggctttca
     3421 tcctggagca gactttgcag tctgtggact gcaacacaac attgccttta tgtgtaactc
     3481 ttggctgaag ctcttacacc aatgctgggg gacatgtacc tcccaggggc ccaggaagac
     3541 tacgggaggc tacaccaacg tcaatcagag gggcctgtgt agctaccgat aagcggaccc
     3601 tcaagagggc attagcaata gtgtttataa ggcccccttg ttaaccctaa acgggtagca
     3661 tatgcttccc gggtagtagt atatactatc cagactaacc ctaattcaat agcatatgtt
     3721 acccaacggg aagcatatgc tatcgaatta gggttagtaa aagggtccta aggaacagcg
     3781 atatctccca ccccatgagc tgtcacggtt ttatttacat ggggtcagga ttccacgagg
     3841 gtagtgaacc attttagtca caagggcagt ggctgaagat caaggagcgg gcagtgaact
     3901 ctcctgaatc ttcgcctgct tcttcattct ccttcgttta gctaatagaa taactgctga
     3961 gttgtgaaca gtaaggtgta tgtgaggtgc tcgaaaacaa ggtttcaggt gacgccccca
     4021 gaataaaatt tggacggggg gttcagtggt ggcattgtgc tatgacacca atataaccct
     4081 cacaaacccc ttgggcaata aatactagtg taggaatgaa acattctgaa tatctttaac
     4141 aatagaaatc catggggtgg ggacaagccg taaagactgg atgtccatct cacacgaatt
     4201 tatggctatg ggcaacacat aatcctagtg caatatgata ctggggttat taagatgtgt
     4261 cccaggcagg gaccaagaca ggtgaaccat gttgttacac tctatttgta acaaggggaa
     4321 agagagtgga cgccgacagc agcggactcc actggttgtc tctaacaccc ccgaaaatta
     4381 aacggggctc cacgccaatg gggcccataa acaaagacaa gtggccactc ttttttttga
     4441 aattgtggag tgggggcacg cgtcagcccc cacacgccgc cctgcggttt tggactgtaa
     4501 aataagggtg taataacttg gctgattgta accccgctaa ccactgcggt caaaccactt
     4561 gcccacaaaa ccactaatgg caccccgggg aatacctgca taagtaggtg ggcgggccaa
     4621 gataggggcg cgattgctgc gatctggagg acaaattaca cacacttgcg cctgagcgcc
     4681 aagcacaggg ttgttggtcc tcatattcac gaggtcgctg agagcacggt gggctaatgt
     4741 tgccatgggt agcatatact acccaaatat ctggatagca tatgctatcc taatctatat
     4801 ctgggtagca taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat
     4861 ctgggtagta tatgctatcc taatttatat ctgggtagca taggctatcc taatctatat
     4921 ctgggtagca tatgctatcc taatctatat ctgggtagta tatgctatcc taatctgtat
     4981 ccgggtagca tatgctatcc taatagagat tagggtagta tatgctatcc taatttatat
     5041 ctgggtagca tatactaccc aaatatctgg atagcatatg ctatcctaat ctatatctgg
     5101 gtagcatatg ctatcctaat ctatatctgg gtagcatagg ctatcctaat ctatatctgg
     5161 gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat ttatatctgg
     5221 gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg
     5281 gtagtatatg ctatcctaat ctgtatccgg gtagcatatg ctatcctcat gataagctgt
     5341 caaacatgag aattttcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt
     5401 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     5461 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     5521 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     5581 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     5641 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     5701 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     5761 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa
     5821 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     5881 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     5941 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     6001 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     6061 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca
     6121 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     6181 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     6241 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     6301 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     6361 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     6421 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     6481 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     6541 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     6601 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     6661 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     6721 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     6781 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     6841 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     6901 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     6961 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     7021 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     7081 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     7141 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     7201 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     7261 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     7321 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     7381 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     7441 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     7501 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     7561 tttcacacag gaaacagcta tgaccatgat tacgccaagc tctagctaga ggtcgaccaa
     7621 ttctcatgtt tgacagctta tcatcgcaga tccgggcaac gttgttgcca ttgctgcagg
     7681 cgcagaactg gtaggtatgg aagatctata cattgaatca atattggcaa ttagccatat
     7741 tagtcattgg ttatatagca taaatcaata ttggctattg gccattgcat acgttgtatc
     7801 tatatcataa tatgtacatt tatattggct catgtccaat atgaccgcca tgtt
//
