LOCUS       sequence-274908-        6603 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 6603)
  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 6603)
  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: D24_Ld_171460
FEATURES             Location/Qualifiers
     source          1..6603
                     /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             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 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 gccaccgtga
     1201 tcagcagctg tgatgtgtgc ggcctgagca tcgtgaagca ggtcatggcc cccagcgaca
     1261 gatccgatgg cggcgtgcca gaggccacag ccgtgcggtc tagccccagc accgccacag
     1321 agacacccat gctggaactg gacgaccggc ctagaggcgc cgctgccggc aaggccaaca
     1381 aatacgtgga agagttctac caggccgctg gccacctgag atgccctcag ctgcagcggt
     1441 acacccagga agaggtcgcc cggcaccgga ccaaggacga cctgtggatc gtggtggaca
     1501 gcaacgtgct gaacgtgtcc gccttcgtgc cccaccatcc tggcggcgac gtgatcctgg
     1561 atggcgtggg cggccaggac atggccacag tgttcgccta cttccacgac ccctccaccg
     1621 tgcggctgat gctgagcttc tgcatcggcc ggatcctgct gcagggcgcg 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 tgaccgccat
     6601 gtt
//
