LOCUS       sequence-275075-        7737 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 7737)
  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 7737)
  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: D22_Ld_141410
FEATURES             Location/Qualifiers
     source          1..7737
                     /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             2901..2918
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2970..3025
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3024..3043)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5261..5279)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5347..5451
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5452..5520,5521..6312)
                     /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(5670..5689)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6483..7071
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6972..6991
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7225..7242
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7359..7380
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7395..7400,7401..7418,7419..7425)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7433..7449
                     /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     7438..7460
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7457..7473
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7457..7473
                     /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 ccgtgtttgg cgtggaagcc gacgacgaca ccatccggct gctgctggcc gctggcgata
     1261 aggccctggg acagggcaga gcccactacg gcgaggccct gagcaagtac accgccgtgc
     1321 tgacccggtg gcccaagaac gagcgggccc tgtacagcag agccgagctg tactccatga
     1381 tgagagagcg ggctgccgcc ctggacgacc tgaatgccct gctgaccatc aacaaggacc
     1441 acccccaggg cctggccctg cggatgagcc tgaacaccca gctgggcaac ctggtggacg
     1501 cccacaggga cggaaagcac ctcgtgcggg tgtacaaagc cctgaacaag cccgacaaga
     1561 gccagcagac cagcgagaaa gtgcggcacc tggaacacta cacagcccgg tggaccgccc
     1621 tgtccgatct gtggtcacag cctgtgggcg cctacaccga cgccgccaac gacgctgtgc
     1681 tgatgcggaa gtaccgcgag tgcgtggacg tgctggccag aatcatccgc gagttcagcg
     1741 tggactccgt ggaactgcgg ctgagaaggg ccgcctgtgc cctggccgcc gatgacaata
     1801 tcgccgccac acaggaactg aaatacgtca cccagcggag cccccagaac ctggacgcca
     1861 ttgctctgaa cgcccaggct ctgcggcggc tgggagccct ggatcagagc atgagcgagc
     1921 tgaggcggtg cctgaatctg gaccccgagt acgccccctg cgccaacctg cacaagctga
     1981 tccggcagca gcagagaatg atcaagagca tcgagaagcg gctgcaggac aagaaattcg
     2041 aggccgtcgt ccggctgatc gccgaggcca gagccgccga acctcacgcc ccttacgagg
     2101 aacagctggc tgcctggcac tgtgaagccc tggtgtccct gagaaacacc gatgagggca
     2161 tcagagtgtg ccagatgctg gtggatagat acgacggcgc caatagccct accgtgttcg
     2221 atgcccacat cagactggcc gaactgcatc tgctggacga taacattgcc gccgctgagg
     2281 ccgctctgca gaaagccaga gagatgcggc agcacgacgg caaggtggaa gagatgagaa
     2341 tgaagattga gaagctgaag cggaccggcg ccagaaagga ctactacaag atcctgggcc
     2401 tgaagaaaac cgccagcgcc caggacatca ggcgggccta cagaaagctg gccaagagca
     2461 gccaccccga ccagctgcgg agcaaggaca tgaccgaccg cgagcgcgag aagcaggaaa
     2521 agatgttccg ggacgtgaac gaggccaaag aggtgctgct cgacgaggaa aagcgggcca
     2581 gatacgataa tggcgaggat gtgaaccagc ctgccggcca gcggggcgag ccattcttcg
     2641 gctctcactt cgctggcggc ttcccaggcg gctttcctgg gggatttccc ggcggattcc
     2701 caggggggtt ccctggcggg tttcctggcg gctttccagg cgggttccac cagcagagag
     2761 atggcggagg ccaccagcag ttctttttct tcaaccggga aaatggcgcg cccggcagcc
     2821 tgcaccacat cctggacgcc cagaagatgc tgtggaacca cagggacagg aacctgcctc
     2881 ctctggcccc tctgggcccc caccaccatc accaccattg agtctagaca cgtgattaat
     2941 taaggatccc ccgacctcga cctctggcta ataaaggaaa tttattttca ttgcaatagt
     3001 gtgttggaat tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttggt
     3061 cgagatccct cggagatctc tagctagagg atcgatcccc gccccggacg aactaaacct
     3121 gactacgaca tctctgcccc ttcttcgcgg ggcagtgcat gtaatccctt cagttggttg
     3181 gtacaacttg ccaactgggc cctgttccac atgtgacacg gggggggacc aaacacaaag
     3241 gggttctctg actgtagttg acatccttat aaatggatgt gcacatttgc caacactgag
     3301 tggctttcat cctggagcag actttgcagt ctgtggactg caacacaaca ttgcctttat
     3361 gtgtaactct tggctgaagc tcttacacca atgctggggg acatgtacct cccaggggcc
     3421 caggaagact acgggaggct acaccaacgt caatcagagg ggcctgtgta gctaccgata
     3481 agcggaccct caagagggca ttagcaatag tgtttataag gcccccttgt taaccctaaa
     3541 cgggtagcat atgcttcccg ggtagtagta tatactatcc agactaaccc taattcaata
     3601 gcatatgtta cccaacggga agcatatgct atcgaattag ggttagtaaa agggtcctaa
     3661 ggaacagcga tatctcccac cccatgagct gtcacggttt tatttacatg gggtcaggat
     3721 tccacgaggg tagtgaacca ttttagtcac aagggcagtg gctgaagatc aaggagcggg
     3781 cagtgaactc tcctgaatct tcgcctgctt cttcattctc cttcgtttag ctaatagaat
     3841 aactgctgag ttgtgaacag taaggtgtat gtgaggtgct cgaaaacaag gtttcaggtg
     3901 acgcccccag aataaaattt ggacgggggg ttcagtggtg gcattgtgct atgacaccaa
     3961 tataaccctc acaaacccct tgggcaataa atactagtgt aggaatgaaa cattctgaat
     4021 atctttaaca atagaaatcc atggggtggg gacaagccgt aaagactgga tgtccatctc
     4081 acacgaattt atggctatgg gcaacacata atcctagtgc aatatgatac tggggttatt
     4141 aagatgtgtc ccaggcaggg accaagacag gtgaaccatg ttgttacact ctatttgtaa
     4201 caaggggaaa gagagtggac gccgacagca gcggactcca ctggttgtct ctaacacccc
     4261 cgaaaattaa acggggctcc acgccaatgg ggcccataaa caaagacaag tggccactct
     4321 tttttttgaa attgtggagt gggggcacgc gtcagccccc acacgccgcc ctgcggtttt
     4381 ggactgtaaa ataagggtgt aataacttgg ctgattgtaa ccccgctaac cactgcggtc
     4441 aaaccacttg cccacaaaac cactaatggc accccgggga atacctgcat aagtaggtgg
     4501 gcgggccaag ataggggcgc gattgctgcg atctggagga caaattacac acacttgcgc
     4561 ctgagcgcca agcacagggt tgttggtcct catattcacg aggtcgctga gagcacggtg
     4621 ggctaatgtt gccatgggta gcatatacta cccaaatatc tggatagcat atgctatcct
     4681 aatctatatc tgggtagcat aggctatcct aatctatatc tgggtagcat atgctatcct
     4741 aatctatatc tgggtagtat atgctatcct aatttatatc tgggtagcat aggctatcct
     4801 aatctatatc tgggtagcat atgctatcct aatctatatc tgggtagtat atgctatcct
     4861 aatctgtatc cgggtagcat atgctatcct aatagagatt agggtagtat atgctatcct
     4921 aatttatatc tgggtagcat atactaccca aatatctgga tagcatatgc tatcctaatc
     4981 tatatctggg tagcatatgc tatcctaatc tatatctggg tagcataggc tatcctaatc
     5041 tatatctggg tagcatatgc tatcctaatc tatatctggg tagtatatgc tatcctaatt
     5101 tatatctggg tagcataggc tatcctaatc tatatctggg tagcatatgc tatcctaatc
     5161 tatatctggg tagtatatgc tatcctaatc tgtatccggg tagcatatgc tatcctcatg
     5221 ataagctgtc aaacatgaga attttcttga agacgaaagg gcctcgtgat acgcctattt
     5281 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     5341 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     5401 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     5461 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     5521 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     5581 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     5641 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac
     5701 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     5761 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     5821 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     5881 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     5941 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca
     6001 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     6061 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     6121 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     6181 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     6241 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     6301 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     6361 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     6421 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     6481 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     6541 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     6601 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     6661 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     6721 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     6781 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     6841 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     6901 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     6961 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     7021 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     7081 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     7141 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     7201 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     7261 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     7321 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     7381 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     7441 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct ctagctagag
     7501 gtcgaccaat tctcatgttt gacagcttat catcgcagat ccgggcaacg ttgttgccat
     7561 tgctgcaggc gcagaactgg taggtatgga agatctatac attgaatcaa tattggcaat
     7621 tagccatatt agtcattggt tatatagcat aaatcaatat tggctattgg ccattgcata
     7681 cgttgtatct atatcataat atgtacattt atattggctc atgtccaata tgaccgc
//
