LOCUS       sequence-262336-        6507 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 6507)
  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 6507)
  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: D43_Ld_300860
FEATURES             Location/Qualifiers
     source          1..6507
                     /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             1671..1688
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1740..1795
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1794..1813)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4031..4049)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4117..4221
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4222..4290,4291..5082)
                     /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(4440..4459)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5253..5841
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5742..5761
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5995..6012
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6129..6150
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6165..6170,6171..6188,6189..6195)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6203..6219
                     /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     6208..6230
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6227..6243
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6227..6243
                     /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 ccgcctcttc tacccctgcc cctaccgacc tgatcaccgt ggccggaaga gtgcacgtgc
     1261 ggaacgaggt ggtgcagtac gtggtcgtgc gggtggtgga cgacaagacc ggggtggtcg
     1321 tcagaagcgc ccctctggac gccaccagaa gcgtgtcctt cgccaatatc gccagcagcg
     1381 ccgtgtccct ggaagccacc gtggaactgc ccgaccacct gttcgagctg gatgccagct
     1441 ccagcgtgct gaagtccgcc atcagcacca gcagctccac cacccccgtg cagctgacca
     1501 tctccacaat cagcaaggcc gagggcgctg ccagaaaggc ccctacacct acagccggca
     1561 gcagcggcgc ttctggcgcg cccggcagcc tgcaccacat cctggacgcc cagaagatgc
     1621 tgtggaacca cagggacagg aacctgcctc ctctggcccc tctgggcccc caccaccatc
     1681 accaccattg agtctagaca cgtgattaat taaggatccc ccgacctcga cctctggcta
     1741 ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg
     1801 aaggacatat gggagggcaa atcatttggt cgagatccct cggagatctc tagctagagg
     1861 atcgatcccc gccccggacg aactaaacct gactacgaca tctctgcccc ttcttcgcgg
     1921 ggcagtgcat gtaatccctt cagttggttg gtacaacttg ccaactgggc cctgttccac
     1981 atgtgacacg gggggggacc aaacacaaag gggttctctg actgtagttg acatccttat
     2041 aaatggatgt gcacatttgc caacactgag tggctttcat cctggagcag actttgcagt
     2101 ctgtggactg caacacaaca ttgcctttat gtgtaactct tggctgaagc tcttacacca
     2161 atgctggggg acatgtacct cccaggggcc caggaagact acgggaggct acaccaacgt
     2221 caatcagagg ggcctgtgta gctaccgata agcggaccct caagagggca ttagcaatag
     2281 tgtttataag gcccccttgt taaccctaaa cgggtagcat atgcttcccg ggtagtagta
     2341 tatactatcc agactaaccc taattcaata gcatatgtta cccaacggga agcatatgct
     2401 atcgaattag ggttagtaaa agggtcctaa ggaacagcga tatctcccac cccatgagct
     2461 gtcacggttt tatttacatg gggtcaggat tccacgaggg tagtgaacca ttttagtcac
     2521 aagggcagtg gctgaagatc aaggagcggg cagtgaactc tcctgaatct tcgcctgctt
     2581 cttcattctc cttcgtttag ctaatagaat aactgctgag ttgtgaacag taaggtgtat
     2641 gtgaggtgct cgaaaacaag gtttcaggtg acgcccccag aataaaattt ggacgggggg
     2701 ttcagtggtg gcattgtgct atgacaccaa tataaccctc acaaacccct tgggcaataa
     2761 atactagtgt aggaatgaaa cattctgaat atctttaaca atagaaatcc atggggtggg
     2821 gacaagccgt aaagactgga tgtccatctc acacgaattt atggctatgg gcaacacata
     2881 atcctagtgc aatatgatac tggggttatt aagatgtgtc ccaggcaggg accaagacag
     2941 gtgaaccatg ttgttacact ctatttgtaa caaggggaaa gagagtggac gccgacagca
     3001 gcggactcca ctggttgtct ctaacacccc cgaaaattaa acggggctcc acgccaatgg
     3061 ggcccataaa caaagacaag tggccactct tttttttgaa attgtggagt gggggcacgc
     3121 gtcagccccc acacgccgcc ctgcggtttt ggactgtaaa ataagggtgt aataacttgg
     3181 ctgattgtaa ccccgctaac cactgcggtc aaaccacttg cccacaaaac cactaatggc
     3241 accccgggga atacctgcat aagtaggtgg gcgggccaag ataggggcgc gattgctgcg
     3301 atctggagga caaattacac acacttgcgc ctgagcgcca agcacagggt tgttggtcct
     3361 catattcacg aggtcgctga gagcacggtg ggctaatgtt gccatgggta gcatatacta
     3421 cccaaatatc tggatagcat atgctatcct aatctatatc tgggtagcat aggctatcct
     3481 aatctatatc tgggtagcat atgctatcct aatctatatc tgggtagtat atgctatcct
     3541 aatttatatc tgggtagcat aggctatcct aatctatatc tgggtagcat atgctatcct
     3601 aatctatatc tgggtagtat atgctatcct aatctgtatc cgggtagcat atgctatcct
     3661 aatagagatt agggtagtat atgctatcct aatttatatc tgggtagcat atactaccca
     3721 aatatctgga tagcatatgc tatcctaatc tatatctggg tagcatatgc tatcctaatc
     3781 tatatctggg tagcataggc tatcctaatc tatatctggg tagcatatgc tatcctaatc
     3841 tatatctggg tagtatatgc tatcctaatt tatatctggg tagcataggc tatcctaatc
     3901 tatatctggg tagcatatgc tatcctaatc tatatctggg tagtatatgc tatcctaatc
     3961 tgtatccggg tagcatatgc tatcctcatg ataagctgtc aaacatgaga attttcttga
     4021 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     4081 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4141 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4201 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     4261 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     4321 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     4381 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     4441 ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata
     4501 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     4561 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     4621 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     4681 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     4741 gacgagcgtg acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact
     4801 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     4861 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     4921 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     4981 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     5041 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     5101 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     5161 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     5221 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     5281 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     5341 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     5401 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     5461 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     5521 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     5581 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     5641 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     5701 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     5761 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     5821 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     5881 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     5941 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     6001 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     6061 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     6121 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     6181 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     6241 gaccatgatt acgccaagct ctagctagag gtcgaccaat tctcatgttt gacagcttat
     6301 catcgcagat ccgggcaacg ttgttgccat tgctgcaggc gcagaactgg taggtatgga
     6361 agatctatac attgaatcaa tattggcaat tagccatatt agtcattggt tatatagcat
     6421 aaatcaatat tggctattgg ccattgcata cgttgtatct atatcataat atgtacattt
     6481 atattggctc atgtccaata tgaccgc
//
