LOCUS       sequence-262363-        7047 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 7047)
  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 7047)
  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: D71_Ld_230810
FEATURES             Location/Qualifiers
     source          1..7047
                     /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             2211..2228
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2280..2335
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2334..2353)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4571..4589)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4657..4761
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4762..4830,4831..5622)
                     /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(4980..4999)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5793..6381
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6282..6301
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6535..6552
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6669..6690
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6705..6710,6711..6728,6729..6735)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6743..6759
                     /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     6748..6770
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6767..6783
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6767..6783
                     /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 aagaggtcgc cgccactgcc acagatgccg ccggaaagcc tgtggctcag cacacgtccg
     1261 ttacaactgc aacctctgag gaagaggcga gggccagcaa cgcagcagtg caggcggcag
     1321 aggctgaagc tgccgcaatg cgccaactgg aagctgagga gcaagcaaga cgcgaagaag
     1381 aggaagagtt ggccgagcgt caacggcgac agcaagaaga agagaggaga agacaggaag
     1441 aagaggaatt ggccgagaga cagcgccgcc aacaagaaga ggaaagaaga agacaggaag
     1501 aagaggagct cgcagagcgc caacgtcggc agcaagagga agaaagggcg aagcggcgca
     1561 ttgatagcgt ggatatgatc cgctatagcg ccgaggaact gccccaagga tgtgctaaaa
     1621 gtatagagga gtacctgttt cgcgtccacg aacaccgtca acacacagcc gctgcagtaa
     1681 gggatgccag gtccgaattg aatgaggcta gggtgcatgc accaaagaaa gtggcggaac
     1741 tggagaaacg gctcgctgct cggcaggatg acctgctggt atacgatatc caaactctgc
     1801 gaatgtacgg cgggtcagtt cccaagacat gtattgccga aagtctggct tggctcgacg
     1861 cacaacctga cgtcagctcc atgagtgagg caaagcaacg gttggtatat ttcttccgac
     1921 tcacttctac ccattgtaga gctctgtacc tcactgcacg aggagtgatg caatatatga
     1981 tagacagcta tacccccgta gtgaagtctt acactaaatc cgtgcagtat tattgtgata
     2041 aagccagtgc gatttacgct gaattgctgc ctgcatcagg caagcggcct gacataccca
     2101 tgatggattt ggccggcgcg cccggcagcc tgcaccacat cctggacgcc cagaagatgc
     2161 tgtggaacca cagggacagg aacctgcctc ctctggcccc tctgggcccc caccaccatc
     2221 accaccattg agtctagaca cgtgattaat taaggatccc ccgacctcga cctctggcta
     2281 ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg
     2341 aaggacatat gggagggcaa atcatttggt cgagatccct cggagatctc tagctagagg
     2401 atcgatcccc gccccggacg aactaaacct gactacgaca tctctgcccc ttcttcgcgg
     2461 ggcagtgcat gtaatccctt cagttggttg gtacaacttg ccaactgggc cctgttccac
     2521 atgtgacacg gggggggacc aaacacaaag gggttctctg actgtagttg acatccttat
     2581 aaatggatgt gcacatttgc caacactgag tggctttcat cctggagcag actttgcagt
     2641 ctgtggactg caacacaaca ttgcctttat gtgtaactct tggctgaagc tcttacacca
     2701 atgctggggg acatgtacct cccaggggcc caggaagact acgggaggct acaccaacgt
     2761 caatcagagg ggcctgtgta gctaccgata agcggaccct caagagggca ttagcaatag
     2821 tgtttataag gcccccttgt taaccctaaa cgggtagcat atgcttcccg ggtagtagta
     2881 tatactatcc agactaaccc taattcaata gcatatgtta cccaacggga agcatatgct
     2941 atcgaattag ggttagtaaa agggtcctaa ggaacagcga tatctcccac cccatgagct
     3001 gtcacggttt tatttacatg gggtcaggat tccacgaggg tagtgaacca ttttagtcac
     3061 aagggcagtg gctgaagatc aaggagcggg cagtgaactc tcctgaatct tcgcctgctt
     3121 cttcattctc cttcgtttag ctaatagaat aactgctgag ttgtgaacag taaggtgtat
     3181 gtgaggtgct cgaaaacaag gtttcaggtg acgcccccag aataaaattt ggacgggggg
     3241 ttcagtggtg gcattgtgct atgacaccaa tataaccctc acaaacccct tgggcaataa
     3301 atactagtgt aggaatgaaa cattctgaat atctttaaca atagaaatcc atggggtggg
     3361 gacaagccgt aaagactgga tgtccatctc acacgaattt atggctatgg gcaacacata
     3421 atcctagtgc aatatgatac tggggttatt aagatgtgtc ccaggcaggg accaagacag
     3481 gtgaaccatg ttgttacact ctatttgtaa caaggggaaa gagagtggac gccgacagca
     3541 gcggactcca ctggttgtct ctaacacccc cgaaaattaa acggggctcc acgccaatgg
     3601 ggcccataaa caaagacaag tggccactct tttttttgaa attgtggagt gggggcacgc
     3661 gtcagccccc acacgccgcc ctgcggtttt ggactgtaaa ataagggtgt aataacttgg
     3721 ctgattgtaa ccccgctaac cactgcggtc aaaccacttg cccacaaaac cactaatggc
     3781 accccgggga atacctgcat aagtaggtgg gcgggccaag ataggggcgc gattgctgcg
     3841 atctggagga caaattacac acacttgcgc ctgagcgcca agcacagggt tgttggtcct
     3901 catattcacg aggtcgctga gagcacggtg ggctaatgtt gccatgggta gcatatacta
     3961 cccaaatatc tggatagcat atgctatcct aatctatatc tgggtagcat aggctatcct
     4021 aatctatatc tgggtagcat atgctatcct aatctatatc tgggtagtat atgctatcct
     4081 aatttatatc tgggtagcat aggctatcct aatctatatc tgggtagcat atgctatcct
     4141 aatctatatc tgggtagtat atgctatcct aatctgtatc cgggtagcat atgctatcct
     4201 aatagagatt agggtagtat atgctatcct aatttatatc tgggtagcat atactaccca
     4261 aatatctgga tagcatatgc tatcctaatc tatatctggg tagcatatgc tatcctaatc
     4321 tatatctggg tagcataggc tatcctaatc tatatctggg tagcatatgc tatcctaatc
     4381 tatatctggg tagtatatgc tatcctaatt tatatctggg tagcataggc tatcctaatc
     4441 tatatctggg tagcatatgc tatcctaatc tatatctggg tagtatatgc tatcctaatc
     4501 tgtatccggg tagcatatgc tatcctcatg ataagctgtc aaacatgaga attttcttga
     4561 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     4621 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4681 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4741 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     4801 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     4861 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     4921 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     4981 ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata
     5041 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     5101 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     5161 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     5221 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     5281 gacgagcgtg acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact
     5341 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     5401 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     5461 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     5521 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     5581 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     5641 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     5701 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     5761 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     5821 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     5881 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     5941 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     6001 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     6061 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     6121 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     6181 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     6241 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     6301 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     6361 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     6421 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     6481 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     6541 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     6601 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     6661 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     6721 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     6781 gaccatgatt acgccaagct ctagctagag gtcgaccaat tctcatgttt gacagcttat
     6841 catcgcagat ccgggcaacg ttgttgccat tgctgcaggc gcagaactgg taggtatgga
     6901 agatctatac attgaatcaa tattggcaat tagccatatt agtcattggt tatatagcat
     6961 aaatcaatat tggctattgg ccattgcata cgttgtatct atatcataat atgtacattt
     7021 atattggctc atgtccaata tgaccgc
//
