LOCUS       sequence-262362-        7251 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 7251)
  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 7251)
  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: D70_Ld_210610
FEATURES             Location/Qualifiers
     source          1..7251
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        7..386
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        387..590
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     541..561
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     587..611
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     1092..1111
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1134..1143
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2415..2432
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2484..2539
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2538..2557)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4775..4793)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4861..4965
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4966..5034,5035..5826)
                     /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(5184..5203)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5997..6585
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6486..6505
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6739..6756
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6873..6894
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6909..6914,6915..6932,6933..6939)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6947..6963
                     /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     6952..6974
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6971..6987
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6971..6987
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 catgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
       61 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      121 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      181 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      241 tacatcaagt gtatcatatg ccaagtccgc cccctattga cgtcaatgac ggtaaatggc
      301 ccgcctggca ttatgcccag tacatgacct tacgggactt tcctacttgg cagtacatct
      361 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacacc aatgggcgtg
      421 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      481 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taataacccc gccccgttga
      541 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga
      601 accgtcagat cctcactctc ttccgcatcg ctgtctgcga gggccagctg ttgggctcgc
      661 ggttgaggac aaactcttcg cggtctttcc agtactcttg gatcggaaac ccgtcggcct
      721 ccgaacggta ctccgccacc gagggacctg agcgagtccg catcgaccgg atcggaaaac
      781 ctctcgagaa aggcgtctaa ccagtcacag tcgcaaggta ggctgagcac cgtggcgggc
      841 ggcagcgggt ggcggtcggg gttgtttctg gcggaggtgc tgctgatgat gtaattaaag
      901 taggcggtct tgagacggcg gatggtcgag gtgaggtgtg gcaggcttga gatccagctg
      961 ttggggtgag tactccctct caaaagcggg cattacttct gcgctaagat tgtcagtttc
     1021 caaaaacgag gaggatttga tattcacctg gcccgatctg gccatacact tgagtgacaa
     1081 tgacatccac tttgcctttc tctccacagg tgtccactcc caggtccaag tttgccacca
     1141 tggagtttca gacccaggta ctcatgtccc tgctgctctg catgtctggt gcggccgccc
     1201 ccagcatgga aaccggaact gttgtggaat ggaagaagaa ggtcggagag cttgtgaaag
     1261 aaagcgaagt tttctgcaca attcagactg acaaagccgt cgtggattac actaatacct
     1321 ttgaatcagg ctacttggcg aagatatatt gtgggaacgg acagagcgct ccagttgcta
     1381 aaactattgc cgttatggtg tcagacgccg cagatgtctc taaagcagac gaatatacac
     1441 ctgaaggcga ggtgcccgca gctgatgctg aggcaccagc agctgcggcg gtggcagcag
     1501 ctccagctgc cggcggagct tcatctgaag tgccggaagg agtcacctgt gaaccagtat
     1561 ttatgcctgc actctccccc agtatggaga caggcacagt cgtggagtgg aagaagaaag
     1621 tcggagaact cgtgaaagag tctgaggtgt tttgtaccat tcaaacggac aaagccgtag
     1681 ttgattacac caacacgttt gaatctggtt atcttgctaa gatctactgt gggaatggtc
     1741 agtccgcacc agtggctaag acaattgccg tgatggtttc caatgccgcc gatgtcgaga
     1801 aagtggctaa ttattatcct gaggatgcag taggcggacc accggcatct gccgctgacc
     1861 caagcgctgc agcagcagcc gcagcatctg caaggccagc tgcttcagct gccagtgcga
     1921 aacactatgg cgggtctttg gacgccgccg tggccgcaag tggaccatct gtggccagaa
     1981 ttgctgctgg cctcgaacca agtgcccttg ctgggattgc tcctagtggg aaaggcggga
     2041 gattcttgaa gagtgatttc agcggccaac ccggattcga ctacaacgat actgcgcccg
     2101 ctcgcgctat gcaacaaaag gctgctcccg cagctgccgc cgatggcgct tcaaaggccg
     2161 ctgcaaaatc agctgcaccc gcggctgtca gcggagatat ttacaacgtg gttcttaagc
     2221 ccggccccgt ttataaaagc gtctcagata ccgcactgct gaagaagctg atgtgttcca
     2281 tgcacatccc aaaaccgaaa agcaagaagg cagccgaagg cgcgcccggc agcctgcacc
     2341 acatcctgga cgcccagaag atgctgtgga accacaggga caggaacctg cctcctctgg
     2401 cccctctggg cccccaccac catcaccacc attgagtcta gacacgtgat taattaagga
     2461 tcccccgacc tcgacctctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg
     2521 gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt tggtcgagat
     2581 ccctcggaga tctctagcta gaggatcgat ccccgccccg gacgaactaa acctgactac
     2641 gacatctctg ccccttcttc gcggggcagt gcatgtaatc ccttcagttg gttggtacaa
     2701 cttgccaact gggccctgtt ccacatgtga cacggggggg gaccaaacac aaaggggttc
     2761 tctgactgta gttgacatcc ttataaatgg atgtgcacat ttgccaacac tgagtggctt
     2821 tcatcctgga gcagactttg cagtctgtgg actgcaacac aacattgcct ttatgtgtaa
     2881 ctcttggctg aagctcttac accaatgctg ggggacatgt acctcccagg ggcccaggaa
     2941 gactacggga ggctacacca acgtcaatca gaggggcctg tgtagctacc gataagcgga
     3001 ccctcaagag ggcattagca atagtgttta taaggccccc ttgttaaccc taaacgggta
     3061 gcatatgctt cccgggtagt agtatatact atccagacta accctaattc aatagcatat
     3121 gttacccaac gggaagcata tgctatcgaa ttagggttag taaaagggtc ctaaggaaca
     3181 gcgatatctc ccaccccatg agctgtcacg gttttattta catggggtca ggattccacg
     3241 agggtagtga accattttag tcacaagggc agtggctgaa gatcaaggag cgggcagtga
     3301 actctcctga atcttcgcct gcttcttcat tctccttcgt ttagctaata gaataactgc
     3361 tgagttgtga acagtaaggt gtatgtgagg tgctcgaaaa caaggtttca ggtgacgccc
     3421 ccagaataaa atttggacgg ggggttcagt ggtggcattg tgctatgaca ccaatataac
     3481 cctcacaaac cccttgggca ataaatacta gtgtaggaat gaaacattct gaatatcttt
     3541 aacaatagaa atccatgggg tggggacaag ccgtaaagac tggatgtcca tctcacacga
     3601 atttatggct atgggcaaca cataatccta gtgcaatatg atactggggt tattaagatg
     3661 tgtcccaggc agggaccaag acaggtgaac catgttgtta cactctattt gtaacaaggg
     3721 gaaagagagt ggacgccgac agcagcggac tccactggtt gtctctaaca cccccgaaaa
     3781 ttaaacgggg ctccacgcca atggggccca taaacaaaga caagtggcca ctcttttttt
     3841 tgaaattgtg gagtgggggc acgcgtcagc ccccacacgc cgccctgcgg ttttggactg
     3901 taaaataagg gtgtaataac ttggctgatt gtaaccccgc taaccactgc ggtcaaacca
     3961 cttgcccaca aaaccactaa tggcaccccg gggaatacct gcataagtag gtgggcgggc
     4021 caagataggg gcgcgattgc tgcgatctgg aggacaaatt acacacactt gcgcctgagc
     4081 gccaagcaca gggttgttgg tcctcatatt cacgaggtcg ctgagagcac ggtgggctaa
     4141 tgttgccatg ggtagcatat actacccaaa tatctggata gcatatgcta tcctaatcta
     4201 tatctgggta gcataggcta tcctaatcta tatctgggta gcatatgcta tcctaatcta
     4261 tatctgggta gtatatgcta tcctaattta tatctgggta gcataggcta tcctaatcta
     4321 tatctgggta gcatatgcta tcctaatcta tatctgggta gtatatgcta tcctaatctg
     4381 tatccgggta gcatatgcta tcctaataga gattagggta gtatatgcta tcctaattta
     4441 tatctgggta gcatatacta cccaaatatc tggatagcat atgctatcct aatctatatc
     4501 tgggtagcat atgctatcct aatctatatc tgggtagcat aggctatcct aatctatatc
     4561 tgggtagcat atgctatcct aatctatatc tgggtagtat atgctatcct aatttatatc
     4621 tgggtagcat aggctatcct aatctatatc tgggtagcat atgctatcct aatctatatc
     4681 tgggtagtat atgctatcct aatctgtatc cgggtagcat atgctatcct catgataagc
     4741 tgtcaaacat gagaattttc ttgaagacga aagggcctcg tgatacgcct atttttatag
     4801 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     4861 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     4921 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     4981 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     5041 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     5101 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     5161 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg
     5221 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     5281 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     5341 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     5401 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     5461 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca
     5521 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     5581 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     5641 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     5701 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     5761 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     5821 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     5881 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     5941 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     6001 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     6061 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     6121 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     6181 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6241 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6301 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6361 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6421 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6481 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6541 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6601 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     6661 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     6721 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     6781 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     6841 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     6901 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     6961 caatttcaca caggaaacag ctatgaccat gattacgcca agctctagct agaggtcgac
     7021 caattctcat gtttgacagc ttatcatcgc agatccgggc aacgttgttg ccattgctgc
     7081 aggcgcagaa ctggtaggta tggaagatct atacattgaa tcaatattgg caattagcca
     7141 tattagtcat tggttatata gcataaatca atattggcta ttggccattg catacgttgt
     7201 atctatatca taatatgtac atttatattg gctcatgtcc aatatgaccg c
//
