LOCUS       sequence-262298-        7800 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 7800)
  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 7800)
  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: D3_Ld_030930
FEATURES             Location/Qualifiers
     source          1..7800
                     /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             2964..2981
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3033..3088
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3087..3106)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5324..5342)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5410..5514
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5515..5583,5584..6375)
                     /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(5733..5752)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6546..7134
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7035..7054
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7288..7305
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7422..7443
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7458..7463,7464..7481,7482..7488)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7496..7512
                     /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     7501..7523
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7520..7536
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7520..7536
                     /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 ccgctgatgc tgtgtccagc cccgtgatcc tgcccagagg ccggtccaac accaaagtga
     1261 ccctgtacgt gcggaccggc agagtgtcca tccccctgga atggatcgac ggcaaagagg
     1321 tgttcttcga gtacaccggc agactgtacg aagtgggcaa cagcggcccc atgctgcctg
     1381 gccccaccct gaaagtgaac cctggcggca gaatcgtgct gaagctggtc aacgacctgg
     1441 gcaaagaggg catggccaac atgaccggcc ggatgaacac cctgcacggc cccaacatca
     1501 ccaacgtgca cttccacggc atgcacagcg accccaagat ggacaacccc ttcaaggtgg
     1561 ccctgcccgg cgagacactg gtgtacacaa tcaacgtgcc ccgggaccac gagcccggcc
     1621 tgcactggta tcacgcccac agccacgggg ccgtgtacta ccacgtgatg ggcggcatgt
     1681 tcggcgccat cgacgtgggc gagggcgact tcaccaagac ccccagacac cccttccacg
     1741 gctgggacag ccagatcctg atggtgcacc tgtaccggct gaacaccagc gagagatgcg
     1801 acggcatgcc catgagcagc ctggacgccg ccatgtccag cctgctgccc ttcgaccccc
     1861 ggatcgtgga ccggaagggc aacgagtacg agatgcccgc cgagctgttc ctggtcaacg
     1921 gccagcaccg gcccaccgtc agagtgaaga ggggccaccc tatgctgctg cggatcggct
     1981 ttgccgccgg aagctgctac atcaacatga gcctgcccaa gcagtgcgcc ttccacatga
     2041 cagccatcga cggcgcccag ctgccccgga ccgtggaagt ggtggaaggc tggctgtact
     2101 tcaccaccgc cacccggcga agcctggtgg tcgtgtgtga cgaagagggc acctaccccg
     2161 tgtcccacac cggcgatccc agcgacgcca tcttctacct gaagtcccgg ggcaagcacc
     2221 agaaaggcgg cagagccgtg gccttccccg tgaccatgca caagtacagc cccggctacc
     2281 tgcagctgaa gggccagaac accttctaca gagagatcag cttcagccag cgggacctgc
     2341 ccatcaccaa gccctactac gtgctgggcc agggccccaa ctgccagagc ctgcagaaca
     2401 gcagcacctg tcactacgag tacttccagg gcgagatcgg cagacggatc gagggctacc
     2461 acggcttcac cgtgcctctg cacgccgtgg tcaccgccag agtgtacggc gaccccaccg
     2521 acaagaggcc ccaccccctg cactttcacg tgaaccactt ccagtttctg agcttcgagc
     2581 ccagacctgg cggcaaccac gagaacgaga caatggctat gtacggcgtg cactccggcg
     2641 agttccggga caccatcccc atcctggacg gcgtgaccac catccggtgg aaggccgcca
     2701 cctatgtggg cgaggtggtg taccactgcc acgccctgca gcacgaggac cggggcatga
     2761 tgatgagcta cctggtgtac agcccccggt ccaaagaggg aaaggccgtc cagagatacg
     2821 agcagagcca gaccggcgcc ttcctgctga cctggcctca cagatctggc gcgcccggca
     2881 gcctgcacca catcctggac gcccagaaga tgctgtggaa ccacagggac aggaacctgc
     2941 ctcctctggc ccctctgggc ccccaccacc atcaccacca ttgagtctag acacgtgatt
     3001 aattaaggat cccccgacct cgacctctgg ctaataaagg aaatttattt tcattgcaat
     3061 agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg caaatcattt
     3121 ggtcgagatc cctcggagat ctctagctag aggatcgatc cccgccccgg acgaactaaa
     3181 cctgactacg acatctctgc cccttcttcg cggggcagtg catgtaatcc cttcagttgg
     3241 ttggtacaac ttgccaactg ggccctgttc cacatgtgac acgggggggg accaaacaca
     3301 aaggggttct ctgactgtag ttgacatcct tataaatgga tgtgcacatt tgccaacact
     3361 gagtggcttt catcctggag cagactttgc agtctgtgga ctgcaacaca acattgcctt
     3421 tatgtgtaac tcttggctga agctcttaca ccaatgctgg gggacatgta cctcccaggg
     3481 gcccaggaag actacgggag gctacaccaa cgtcaatcag aggggcctgt gtagctaccg
     3541 ataagcggac cctcaagagg gcattagcaa tagtgtttat aaggccccct tgttaaccct
     3601 aaacgggtag catatgcttc ccgggtagta gtatatacta tccagactaa ccctaattca
     3661 atagcatatg ttacccaacg ggaagcatat gctatcgaat tagggttagt aaaagggtcc
     3721 taaggaacag cgatatctcc caccccatga gctgtcacgg ttttatttac atggggtcag
     3781 gattccacga gggtagtgaa ccattttagt cacaagggca gtggctgaag atcaaggagc
     3841 gggcagtgaa ctctcctgaa tcttcgcctg cttcttcatt ctccttcgtt tagctaatag
     3901 aataactgct gagttgtgaa cagtaaggtg tatgtgaggt gctcgaaaac aaggtttcag
     3961 gtgacgcccc cagaataaaa tttggacggg gggttcagtg gtggcattgt gctatgacac
     4021 caatataacc ctcacaaacc ccttgggcaa taaatactag tgtaggaatg aaacattctg
     4081 aatatcttta acaatagaaa tccatggggt ggggacaagc cgtaaagact ggatgtccat
     4141 ctcacacgaa tttatggcta tgggcaacac ataatcctag tgcaatatga tactggggtt
     4201 attaagatgt gtcccaggca gggaccaaga caggtgaacc atgttgttac actctatttg
     4261 taacaagggg aaagagagtg gacgccgaca gcagcggact ccactggttg tctctaacac
     4321 ccccgaaaat taaacggggc tccacgccaa tggggcccat aaacaaagac aagtggccac
     4381 tctttttttt gaaattgtgg agtgggggca cgcgtcagcc cccacacgcc gccctgcggt
     4441 tttggactgt aaaataaggg tgtaataact tggctgattg taaccccgct aaccactgcg
     4501 gtcaaaccac ttgcccacaa aaccactaat ggcaccccgg ggaatacctg cataagtagg
     4561 tgggcgggcc aagatagggg cgcgattgct gcgatctgga ggacaaatta cacacacttg
     4621 cgcctgagcg ccaagcacag ggttgttggt cctcatattc acgaggtcgc tgagagcacg
     4681 gtgggctaat gttgccatgg gtagcatata ctacccaaat atctggatag catatgctat
     4741 cctaatctat atctgggtag cataggctat cctaatctat atctgggtag catatgctat
     4801 cctaatctat atctgggtag tatatgctat cctaatttat atctgggtag cataggctat
     4861 cctaatctat atctgggtag catatgctat cctaatctat atctgggtag tatatgctat
     4921 cctaatctgt atccgggtag catatgctat cctaatagag attagggtag tatatgctat
     4981 cctaatttat atctgggtag catatactac ccaaatatct ggatagcata tgctatccta
     5041 atctatatct gggtagcata tgctatccta atctatatct gggtagcata ggctatccta
     5101 atctatatct gggtagcata tgctatccta atctatatct gggtagtata tgctatccta
     5161 atttatatct gggtagcata ggctatccta atctatatct gggtagcata tgctatccta
     5221 atctatatct gggtagtata tgctatccta atctgtatcc gggtagcata tgctatcctc
     5281 atgataagct gtcaaacatg agaattttct tgaagacgaa agggcctcgt gatacgccta
     5341 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     5401 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     5461 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     5521 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     5581 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     5641 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     5701 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt
     5761 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     5821 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     5881 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     5941 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     6001 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca
     6061 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     6121 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     6181 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     6241 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     6301 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     6361 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     6421 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     6481 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     6541 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     6601 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     6661 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     6721 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     6781 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6841 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     6901 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     6961 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     7021 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     7081 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     7141 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     7201 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     7261 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     7321 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     7381 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     7441 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     7501 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gctctagcta
     7561 gaggtcgacc aattctcatg tttgacagct tatcatcgca gatccgggca acgttgttgc
     7621 cattgctgca ggcgcagaac tggtaggtat ggaagatcta tacattgaat caatattggc
     7681 aattagccat attagtcatt ggttatatag cataaatcaa tattggctat tggccattgc
     7741 atacgttgta tctatatcat aatatgtaca tttatattgg ctcatgtcca atatgaccgc
//
