LOCUS       sequence-262388-        9354 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 9354)
  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 9354)
  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: D96_Ld_250480
FEATURES             Location/Qualifiers
     source          1..9354
                     /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             4518..4535
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4587..4642
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4641..4660)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6878..6896)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6964..7068
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7069..7137,7138..7929)
                     /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(7287..7306)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8100..8688
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8589..8608
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8842..8859
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8976..8997
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9012..9017,9018..9035,9036..9042)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9050..9066
                     /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     9055..9077
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9074..9090
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9074..9090
                     /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 gcggccgccg
     1201 ctagagctgc tgctgcagga tctgcttgtg gatctctggc ccacgtgtac tacctgcctg
     1261 acggccagga tttcagaggc accgtgcatg tgacagaagg cggcatccct tgccagctga
     1321 gaagcagcca gacacctaat agccacgagg tgccactgga ttctgccaca ggcgtgggca
     1381 accacaacta ctgtagagat gctggcggac tgggcagacc tggctgtttt gctgctacac
     1441 ctgtgggcgc ctatcagctg tgtgccgtta gagccgtgtg taccgctaga agcctggctg
     1501 cccctacagt gcagttttgg cctgtgtctg gaacaaagag agccgccgac ggaagcagct
     1561 cttacctggc cattagctgt caccccagac cttgcagcat ccactatgtg ctgggagccg
     1621 aagccattgc ctctgccaga tctccagtgt acacacagcc tctgctgctg gaacagagca
     1681 ccaccgtgaa aacctacgtc agatacgaga cagccgagcc tatgagagcc caggccagat
     1741 atacagtgac ccctgccgct acatctaccc cagccgcctc tatccagttt atcccttccg
     1801 acgccgtggt gtactctcac cccatctttg tggaactgca gggcatcacc aatgcctccg
     1861 aggtgctgct gttctgggcc gacgatttca tgaaccccac cacatacacc ggcggagcct
     1921 tcagagtgga aagatctaca tctgtggtgg ccgtggtcaa cagcacctac ctggtgtctg
     1981 ccagctacaa gctggacatc acagcccctc cactgaacgt gtaccctcca tctggcacat
     2041 atgtcggcgg cgtgacagtg ctcgttagcg atcctcagcc tcctgccacc tactatatgt
     2101 acgtgaacca gagcaccgac tggcagtctc tgggcacact gctgtttcca tggacagcca
     2161 tcggcaccag cgagctggat attagagcac tgcacgtgga aggcatcgag agcgccgaga
     2221 aggtggtgta tgaagtggtg cctgccacac ctcctaccat cctgcctgat gcctctatcg
     2281 cccaccatag acctgccaac gtgacctgta cagccccact gggaacacca gtggctctgt
     2341 ctgtgtacag agatgccgtg atgcaggatg ccgtggccga aggcgttttc tctgctgtgc
     2401 tttctgagcc tggcgcctac accgtgtctt gcacatatgc cgatgacctg cacgccacac
     2461 acaccacaat ggctattctg cagctggtgg ctgtgcccat gcagcctcca acttttaccc
     2521 ctgcctgcgg cacctctttt cccgccattg ctctgtctct gcagatgcag ctgaatgtgt
     2581 ccaacctgct gcctggcgat ctggccacat cttggtctat ttctgcttct gccggcggag
     2641 gcgccagaat tcctatgatc accagatgcg ccgacgacga cggcgtgttc gaatacgatg
     2701 tgctgcagag caagagcatc caagagccta ccaccatgga agtgtacagc accgctcgga
     2761 gcctgaatcc tctggaagcc gattctcccc ggaccagctg tacctactct ctgtatcctc
     2821 tgggcgccag cgccacacct tacttctctg ccattcctga ctgtgccacc acatctgccc
     2881 tggcctctcc atcttgtatc atggccctga agctccagct ggcctcctgt ctgcacttct
     2941 actctaccga gctgctgagc attgacgctc tgggacctat cgtggccatg agaatgacag
     3001 gactggccgc tgccatccag accgacatgt ataccagaat cggcgagtgt ctgacaggcc
     3061 tgcagcatag aggcgttctg tccgacgtgc ggaatgagac atctggcgag atcatgctgg
     3121 ccacaagctg gcatgtggcc actcctacaa gaatggccgc tcaggtgtac acaggactgc
     3181 ccatcacagt ccaggtgtcc ggctttcata cagacgctgg cgcttatcac ctcgtgcgga
     3241 gcgattactc ttgcgaggat atcggagtgc tgcccgaggc agttctggaa acccctccat
     3301 atcatcctac agccgccgga agctctagag tgtctcctag cacagacggc agcacatacc
     3361 tgacctttag agtgcctatc gccggccagt ataagctgtg cagctacatc agcaacgccc
     3421 tgtatgaggt gcccttcagc tttagcacag gcggagcagc tgcacccgcc gtgaatcagt
     3481 atctggatgt ggttgtgcag ccctggccaa cagttgatgc cgtgcctgct gaagaatgcg
     3541 gaggactggt gcctcctgac aaagaggatg tcgcctttag cctgtctatg gccgctgtgc
     3601 cagcaggcgc ttatgccagc ctggcctaca gctttaatag cgccggatgg ggcagcgtgc
     3661 cattcgctat tgatgctgcc acacagcttg gcggcgctgc ctctctttct atcggccctc
     3721 tgaaccggta caccagacca ctggaagtgc tggatgcttc tctgggccct acaacaggcg
     3781 ccgctcagac ctgcatcttt ttcgtgtctg ccgccaacag cagcgtgcag agagatgaga
     3841 tcacctacct cttctacaag atgaactcca gcgaggtgcc cgccgatctt acaggcgtta
     3901 tgctgctgct ccaaggccag tttcagcccg ccgagatgat cgtgatcgac gtgtacgaga
     3961 acgtgaccct gaatccaacc accgccaaga agtctagcag cctgcagctc ctgagaagag
     4021 tgacagccag acctagcgcc gctacagcat ttgctgctgc ccttccagat gccatccttg
     4081 gattgcctgc cggcgctcag cacatccctt atgtgatcca tgtgaccctg gacggcgtgc
     4141 accttacagc agctccatct actgtggccc tgtctcctct ggctctggcc atggaagcct
     4201 ctacagcttg tcctagcgga ctggctctga agggccagtg cctgtgtaga aacaagcgga
     4261 ccaaagtgat cagcgcctgc ggaaccgcca tcgaggattc tggatcttct gatgccggcg
     4321 agcctcagtc tagaagctct gccgatagct ctagctgctc caccaccgat agcagcgacg
     4381 acaccgatac caagacagtg tccctgggac agagactggc tggcgcgccc ggcagcctgc
     4441 accacatcct ggacgcccag aagatgctgt ggaaccacag ggacaggaac ctgcctcctc
     4501 tggcccctct gggcccccac caccatcacc accattgagt ctagacacgt gattaattaa
     4561 ggatcccccg acctcgacct ctggctaata aaggaaattt attttcattg caatagtgtg
     4621 ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc atttggtcga
     4681 gatccctcgg agatctctag ctagaggatc gatccccgcc ccggacgaac taaacctgac
     4741 tacgacatct ctgccccttc ttcgcggggc agtgcatgta atcccttcag ttggttggta
     4801 caacttgcca actgggccct gttccacatg tgacacgggg ggggaccaaa cacaaagggg
     4861 ttctctgact gtagttgaca tccttataaa tggatgtgca catttgccaa cactgagtgg
     4921 ctttcatcct ggagcagact ttgcagtctg tggactgcaa cacaacattg cctttatgtg
     4981 taactcttgg ctgaagctct tacaccaatg ctgggggaca tgtacctccc aggggcccag
     5041 gaagactacg ggaggctaca ccaacgtcaa tcagaggggc ctgtgtagct accgataagc
     5101 ggaccctcaa gagggcatta gcaatagtgt ttataaggcc cccttgttaa ccctaaacgg
     5161 gtagcatatg cttcccgggt agtagtatat actatccaga ctaaccctaa ttcaatagca
     5221 tatgttaccc aacgggaagc atatgctatc gaattagggt tagtaaaagg gtcctaagga
     5281 acagcgatat ctcccacccc atgagctgtc acggttttat ttacatgggg tcaggattcc
     5341 acgagggtag tgaaccattt tagtcacaag ggcagtggct gaagatcaag gagcgggcag
     5401 tgaactctcc tgaatcttcg cctgcttctt cattctcctt cgtttagcta atagaataac
     5461 tgctgagttg tgaacagtaa ggtgtatgtg aggtgctcga aaacaaggtt tcaggtgacg
     5521 cccccagaat aaaatttgga cggggggttc agtggtggca ttgtgctatg acaccaatat
     5581 aaccctcaca aaccccttgg gcaataaata ctagtgtagg aatgaaacat tctgaatatc
     5641 tttaacaata gaaatccatg gggtggggac aagccgtaaa gactggatgt ccatctcaca
     5701 cgaatttatg gctatgggca acacataatc ctagtgcaat atgatactgg ggttattaag
     5761 atgtgtccca ggcagggacc aagacaggtg aaccatgttg ttacactcta tttgtaacaa
     5821 ggggaaagag agtggacgcc gacagcagcg gactccactg gttgtctcta acacccccga
     5881 aaattaaacg gggctccacg ccaatggggc ccataaacaa agacaagtgg ccactctttt
     5941 ttttgaaatt gtggagtggg ggcacgcgtc agcccccaca cgccgccctg cggttttgga
     6001 ctgtaaaata agggtgtaat aacttggctg attgtaaccc cgctaaccac tgcggtcaaa
     6061 ccacttgccc acaaaaccac taatggcacc ccggggaata cctgcataag taggtgggcg
     6121 ggccaagata ggggcgcgat tgctgcgatc tggaggacaa attacacaca cttgcgcctg
     6181 agcgccaagc acagggttgt tggtcctcat attcacgagg tcgctgagag cacggtgggc
     6241 taatgttgcc atgggtagca tatactaccc aaatatctgg atagcatatg ctatcctaat
     6301 ctatatctgg gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat
     6361 ctatatctgg gtagtatatg ctatcctaat ttatatctgg gtagcatagg ctatcctaat
     6421 ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat
     6481 ctgtatccgg gtagcatatg ctatcctaat agagattagg gtagtatatg ctatcctaat
     6541 ttatatctgg gtagcatata ctacccaaat atctggatag catatgctat cctaatctat
     6601 atctgggtag catatgctat cctaatctat atctgggtag cataggctat cctaatctat
     6661 atctgggtag catatgctat cctaatctat atctgggtag tatatgctat cctaatttat
     6721 atctgggtag cataggctat cctaatctat atctgggtag catatgctat cctaatctat
     6781 atctgggtag tatatgctat cctaatctgt atccgggtag catatgctat cctcatgata
     6841 agctgtcaaa catgagaatt ttcttgaaga cgaaagggcc tcgtgatacg cctattttta
     6901 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     6961 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     7021 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     7081 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     7141 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     7201 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     7261 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc
     7321 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     7381 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     7441 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     7501 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     7561 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg
     7621 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     7681 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     7741 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     7801 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     7861 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     7921 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     7981 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     8041 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     8101 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     8161 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     8221 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     8281 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     8341 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     8401 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     8461 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     8521 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     8581 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     8641 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     8701 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     8761 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     8821 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     8881 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     8941 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     9001 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     9061 taacaatttc acacaggaaa cagctatgac catgattacg ccaagctcta gctagaggtc
     9121 gaccaattct catgtttgac agcttatcat cgcagatccg ggcaacgttg ttgccattgc
     9181 tgcaggcgca gaactggtag gtatggaaga tctatacatt gaatcaatat tggcaattag
     9241 ccatattagt cattggttat atagcataaa tcaatattgg ctattggcca ttgcatacgt
     9301 tgtatctata tcataatatg tacatttata ttggctcatg tccaatatga ccgc
//
