LOCUS       sequence-262368-        6597 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 6597)
  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 6597)
  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: D76_Ld_303190
FEATURES             Location/Qualifiers
     source          1..6597
                     /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             1761..1778
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1830..1885
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1884..1903)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4121..4139)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4207..4311
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4312..4380,4381..5172)
                     /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(4530..4549)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5343..5931
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5832..5851
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6085..6102
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6219..6240
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6255..6260,6261..6278,6279..6285)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6293..6309
                     /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     6298..6320
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6317..6333
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6317..6333
                     /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 aagttgcgca ggaagccgag atcaatcctc atcccttgct cttcgtatca aagaccacat
     1261 ccagcgatga tatagtgttg gggagctccg ttgaagtggt cgtgaccgtt actaactacg
     1321 gacagtcacc cgccttcgat gtgcagattt ccgatctcct tgaggatgga agtctgcaga
     1381 gcaagagcat tgcctacctg ccatacggag ctagtgagac cctgagatat accgtgactc
     1441 cgacagcgct tggtaattat gctgtgagtg tggcagaggt tacctacaac gtcgagcagg
     1501 gtaatactgc gacggctaga aaggctctct caaacttgat ccgcgaagga gaggcttatt
     1561 attacggtga gggcgttgac gatgagtcat ttcgcggcgt tatcagcgta ctcaccaggg
     1621 accggtacga tagactgcac gctcgatata taaaggaatc tactggcgcg cccggcagcc
     1681 tgcaccacat cctggacgcc cagaagatgc tgtggaacca cagggacagg aacctgcctc
     1741 ctctggcccc tctgggcccc caccaccatc accaccattg agtctagaca cgtgattaat
     1801 taaggatccc ccgacctcga cctctggcta ataaaggaaa tttattttca ttgcaatagt
     1861 gtgttggaat tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttggt
     1921 cgagatccct cggagatctc tagctagagg atcgatcccc gccccggacg aactaaacct
     1981 gactacgaca tctctgcccc ttcttcgcgg ggcagtgcat gtaatccctt cagttggttg
     2041 gtacaacttg ccaactgggc cctgttccac atgtgacacg gggggggacc aaacacaaag
     2101 gggttctctg actgtagttg acatccttat aaatggatgt gcacatttgc caacactgag
     2161 tggctttcat cctggagcag actttgcagt ctgtggactg caacacaaca ttgcctttat
     2221 gtgtaactct tggctgaagc tcttacacca atgctggggg acatgtacct cccaggggcc
     2281 caggaagact acgggaggct acaccaacgt caatcagagg ggcctgtgta gctaccgata
     2341 agcggaccct caagagggca ttagcaatag tgtttataag gcccccttgt taaccctaaa
     2401 cgggtagcat atgcttcccg ggtagtagta tatactatcc agactaaccc taattcaata
     2461 gcatatgtta cccaacggga agcatatgct atcgaattag ggttagtaaa agggtcctaa
     2521 ggaacagcga tatctcccac cccatgagct gtcacggttt tatttacatg gggtcaggat
     2581 tccacgaggg tagtgaacca ttttagtcac aagggcagtg gctgaagatc aaggagcggg
     2641 cagtgaactc tcctgaatct tcgcctgctt cttcattctc cttcgtttag ctaatagaat
     2701 aactgctgag ttgtgaacag taaggtgtat gtgaggtgct cgaaaacaag gtttcaggtg
     2761 acgcccccag aataaaattt ggacgggggg ttcagtggtg gcattgtgct atgacaccaa
     2821 tataaccctc acaaacccct tgggcaataa atactagtgt aggaatgaaa cattctgaat
     2881 atctttaaca atagaaatcc atggggtggg gacaagccgt aaagactgga tgtccatctc
     2941 acacgaattt atggctatgg gcaacacata atcctagtgc aatatgatac tggggttatt
     3001 aagatgtgtc ccaggcaggg accaagacag gtgaaccatg ttgttacact ctatttgtaa
     3061 caaggggaaa gagagtggac gccgacagca gcggactcca ctggttgtct ctaacacccc
     3121 cgaaaattaa acggggctcc acgccaatgg ggcccataaa caaagacaag tggccactct
     3181 tttttttgaa attgtggagt gggggcacgc gtcagccccc acacgccgcc ctgcggtttt
     3241 ggactgtaaa ataagggtgt aataacttgg ctgattgtaa ccccgctaac cactgcggtc
     3301 aaaccacttg cccacaaaac cactaatggc accccgggga atacctgcat aagtaggtgg
     3361 gcgggccaag ataggggcgc gattgctgcg atctggagga caaattacac acacttgcgc
     3421 ctgagcgcca agcacagggt tgttggtcct catattcacg aggtcgctga gagcacggtg
     3481 ggctaatgtt gccatgggta gcatatacta cccaaatatc tggatagcat atgctatcct
     3541 aatctatatc tgggtagcat aggctatcct aatctatatc tgggtagcat atgctatcct
     3601 aatctatatc tgggtagtat atgctatcct aatttatatc tgggtagcat aggctatcct
     3661 aatctatatc tgggtagcat atgctatcct aatctatatc tgggtagtat atgctatcct
     3721 aatctgtatc cgggtagcat atgctatcct aatagagatt agggtagtat atgctatcct
     3781 aatttatatc tgggtagcat atactaccca aatatctgga tagcatatgc tatcctaatc
     3841 tatatctggg tagcatatgc tatcctaatc tatatctggg tagcataggc tatcctaatc
     3901 tatatctggg tagcatatgc tatcctaatc tatatctggg tagtatatgc tatcctaatt
     3961 tatatctggg tagcataggc tatcctaatc tatatctggg tagcatatgc tatcctaatc
     4021 tatatctggg tagtatatgc tatcctaatc tgtatccggg tagcatatgc tatcctcatg
     4081 ataagctgtc aaacatgaga attttcttga agacgaaagg gcctcgtgat acgcctattt
     4141 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     4201 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     4261 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     4321 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     4381 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     4441 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     4501 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac
     4561 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     4621 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     4681 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     4741 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     4801 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca
     4861 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     4921 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     4981 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     5041 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     5101 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     5161 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     5221 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     5281 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     5341 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     5401 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     5461 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     5521 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     5581 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     5641 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     5701 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     5761 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     5821 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     5881 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     5941 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     6001 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     6061 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     6121 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     6181 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     6241 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     6301 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct ctagctagag
     6361 gtcgaccaat tctcatgttt gacagcttat catcgcagat ccgggcaacg ttgttgccat
     6421 tgctgcaggc gcagaactgg taggtatgga agatctatac attgaatcaa tattggcaat
     6481 tagccatatt agtcattggt tatatagcat aaatcaatat tggctattgg ccattgcata
     6541 cgttgtatct atatcataat atgtacattt atattggctc atgtccaata tgaccgc
//
