LOCUS       sequence-262333-        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        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             3018..3035
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3087..3142
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3141..3160)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5378..5396)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5464..5568
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5569..5637,5638..6429)
                     /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(5787..5806)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6600..7188
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7089..7108
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7342..7359
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7476..7497
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7512..7517,7518..7535,7536..7542)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7550..7566
                     /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     7555..7577
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7574..7590
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7574..7590
                     /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 gcggccgcca
     1201 ccctgcctca gcaggaagcc cctcccgtga tggaacagcc taccgccgag cctgccgagg
     1261 ccgatggacc tgctagagga cctgcccctc cacccccatc tggcagacca gcccctaatg
     1321 cccctggcgc ctctccccct gccatcggcg attctggcga aacccctgcc cagggcgaga
     1381 cactggaccc catcttcgag gaaatcacca gcctgttcgc cagccggaac cccaagcacg
     1441 ccctgcaggg actgagcgac ggcctgagaa atgccgccac aggcgtggga ctgggcctgg
     1501 ctggcgtggt ggctggaacc tacggcggag ccaaagaggg cggcctgggc ggcatggcca
     1561 aaggctttgg agccggcgtg gccggctttg tgggcctgac aggatatggc gcctacgcag
     1621 gcgtgcggca gatcgtgcgg ggcgtgggca ataccagatc cgccgtgtcc gaggtgtcca
     1681 agggcgaggc ctactgggac agcaacgccc aggcttgggt ccgagtgaac ctggcctgcg
     1741 acttcgagcg gctgccccct accgatgagg atctgctggg agaggcccgg aaggcctacg
     1801 agaaggccaa gaaggacggc acactgccca caaccctgcc cgccatgacc cccaatgata
     1861 gcgccgaggg cggagaagcc agcgccagct ctgctacaac aggcgctggc gccgctagcc
     1921 tgccccaggg aaccgatacc ggcgagacta agcacgctgc cgccgaaccc gtgaactact
     1981 acgccttcct gggcgtggaa agcaccgcca ccagcagcga gatcagaaag gcctataccc
     2041 gcaaggccct ggaaatgcac cccgacaaga accctaacga ccccaacgcc accatcaagt
     2101 tccaggaact gaacaagatc tacaacgtgc tgagccacga ggacacccgg gccacctacg
     2161 accagtacgg caccgtggac cctatgaacg tgcccgagat gaccggcaac cccatgaagg
     2221 aactgctcgg agccgccttt ctggaagccc tcgtgggccc cctgcacttc tttctggtgt
     2281 tcgagggggg cgtgctgttc acagccgaga tgcagagaga gctgcacgcc agacggcggc
     2341 tgcgggtggc caaaaacctg atcagctggc tggacaacga cgtgtccggc ttcgagagcg
     2401 cccagctggc cctgagggat gccgtgtcta cagccctggg ccccgtgttc gtgtcctacg
     2461 tggccgagga ataccacctg gccagcagac agcagctgca tggctccagc tggaagagag
     2521 agatggacag ctggtactcc tcctgggcca catccgccag cagcctgtgg cattggacca
     2581 ccatgggcgc cagaaccgcc aggcgggcct tcgtggacaa gaatctgggc gaagaggaca
     2641 tcctgagagt gctggccgtg gccaacgagc gggacgtccg ccagattgtg ctgcaggcct
     2701 gccggctggt gctgtttgac atgagcgtga ccccagccca gcggcagcag cgggccatga
     2761 gactggaaga tctgtctgtg atggctatga aggaagtggc cctcgaggtc gccagcagag
     2821 agcggattgg agccagcgaa gccacagccg ccgctgccga gaagggcgcc acaacaaaga
     2881 cacccgccga caccagctcc gtgaccgccc cacctctgtc tggcgcgccc ggcagcctgc
     2941 accacatcct ggacgcccag aagatgctgt ggaaccacag ggacaggaac ctgcctcctc
     3001 tggcccctct gggcccccac caccatcacc accattgagt ctagacacgt gattaattaa
     3061 ggatcccccg acctcgacct ctggctaata aaggaaattt attttcattg caatagtgtg
     3121 ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc atttggtcga
     3181 gatccctcgg agatctctag ctagaggatc gatccccgcc ccggacgaac taaacctgac
     3241 tacgacatct ctgccccttc ttcgcggggc agtgcatgta atcccttcag ttggttggta
     3301 caacttgcca actgggccct gttccacatg tgacacgggg ggggaccaaa cacaaagggg
     3361 ttctctgact gtagttgaca tccttataaa tggatgtgca catttgccaa cactgagtgg
     3421 ctttcatcct ggagcagact ttgcagtctg tggactgcaa cacaacattg cctttatgtg
     3481 taactcttgg ctgaagctct tacaccaatg ctgggggaca tgtacctccc aggggcccag
     3541 gaagactacg ggaggctaca ccaacgtcaa tcagaggggc ctgtgtagct accgataagc
     3601 ggaccctcaa gagggcatta gcaatagtgt ttataaggcc cccttgttaa ccctaaacgg
     3661 gtagcatatg cttcccgggt agtagtatat actatccaga ctaaccctaa ttcaatagca
     3721 tatgttaccc aacgggaagc atatgctatc gaattagggt tagtaaaagg gtcctaagga
     3781 acagcgatat ctcccacccc atgagctgtc acggttttat ttacatgggg tcaggattcc
     3841 acgagggtag tgaaccattt tagtcacaag ggcagtggct gaagatcaag gagcgggcag
     3901 tgaactctcc tgaatcttcg cctgcttctt cattctcctt cgtttagcta atagaataac
     3961 tgctgagttg tgaacagtaa ggtgtatgtg aggtgctcga aaacaaggtt tcaggtgacg
     4021 cccccagaat aaaatttgga cggggggttc agtggtggca ttgtgctatg acaccaatat
     4081 aaccctcaca aaccccttgg gcaataaata ctagtgtagg aatgaaacat tctgaatatc
     4141 tttaacaata gaaatccatg gggtggggac aagccgtaaa gactggatgt ccatctcaca
     4201 cgaatttatg gctatgggca acacataatc ctagtgcaat atgatactgg ggttattaag
     4261 atgtgtccca ggcagggacc aagacaggtg aaccatgttg ttacactcta tttgtaacaa
     4321 ggggaaagag agtggacgcc gacagcagcg gactccactg gttgtctcta acacccccga
     4381 aaattaaacg gggctccacg ccaatggggc ccataaacaa agacaagtgg ccactctttt
     4441 ttttgaaatt gtggagtggg ggcacgcgtc agcccccaca cgccgccctg cggttttgga
     4501 ctgtaaaata agggtgtaat aacttggctg attgtaaccc cgctaaccac tgcggtcaaa
     4561 ccacttgccc acaaaaccac taatggcacc ccggggaata cctgcataag taggtgggcg
     4621 ggccaagata ggggcgcgat tgctgcgatc tggaggacaa attacacaca cttgcgcctg
     4681 agcgccaagc acagggttgt tggtcctcat attcacgagg tcgctgagag cacggtgggc
     4741 taatgttgcc atgggtagca tatactaccc aaatatctgg atagcatatg ctatcctaat
     4801 ctatatctgg gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat
     4861 ctatatctgg gtagtatatg ctatcctaat ttatatctgg gtagcatagg ctatcctaat
     4921 ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat
     4981 ctgtatccgg gtagcatatg ctatcctaat agagattagg gtagtatatg ctatcctaat
     5041 ttatatctgg gtagcatata ctacccaaat atctggatag catatgctat cctaatctat
     5101 atctgggtag catatgctat cctaatctat atctgggtag cataggctat cctaatctat
     5161 atctgggtag catatgctat cctaatctat atctgggtag tatatgctat cctaatttat
     5221 atctgggtag cataggctat cctaatctat atctgggtag catatgctat cctaatctat
     5281 atctgggtag tatatgctat cctaatctgt atccgggtag catatgctat cctcatgata
     5341 agctgtcaaa catgagaatt ttcttgaaga cgaaagggcc tcgtgatacg cctattttta
     5401 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     5461 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     5521 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     5581 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     5641 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     5701 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     5761 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc
     5821 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     5881 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     5941 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     6001 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     6061 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg
     6121 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     6181 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     6241 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     6301 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     6361 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     6421 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     6481 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     6541 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     6601 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     6661 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     6721 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     6781 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     6841 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     6901 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     6961 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     7021 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     7081 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     7141 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     7201 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     7261 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     7321 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     7381 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     7441 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     7501 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     7561 taacaatttc acacaggaaa cagctatgac catgattacg ccaagctcta gctagaggtc
     7621 gaccaattct catgtttgac agcttatcat cgcagatccg ggcaacgttg ttgccattgc
     7681 tgcaggcgca gaactggtag gtatggaaga tctatacatt gaatcaatat tggcaattag
     7741 ccatattagt cattggttat atagcataaa tcaatattgg ctattggcca ttgcatacgt
     7801 tgtatctata tcataatatg tacatttata ttggctcatg tccaatatga ccgc
//
