LOCUS       sequence-262316-        7221 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 7221)
  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 7221)
  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: D21_Ld_141150
FEATURES             Location/Qualifiers
     source          1..7221
                     /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             2385..2402
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2454..2509
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2508..2527)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4745..4763)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4831..4935
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4936..5004,5005..5796)
                     /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(5154..5173)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5967..6555
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6456..6475
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6709..6726
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6843..6864
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6879..6884,6885..6902,6903..6909)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6917..6933
                     /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     6922..6944
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6941..6957
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6941..6957
                     /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 ccagcagctt ttgcctgaga ggcagagaca cccacggccc cagactgagc tgtgacgccc
     1261 acctgacagt gtccgccgtg gccctgttct actacggcag cagcgagatc aacagatgcg
     1321 ccgtgcagag cagagccacc gccgtgcctg gcgtgctgat ggtccgagcc gccacagccg
     1381 aaagcagcgc tggcggctct gaggaagaag ccggcggaac agacgcctgg ggcacaatcg
     1441 ccatgaagcg ggccctgaac tacgacagcg gcctgaagcc cttcatgacc ccctggcggg
     1501 aagtgcggag agccctgaga gatgccggct gcttcagcac cctggccctg cctagactga
     1561 ccctgcccac cgatagagcc agccccagag tgggcgacgt gtccccagaa tccagcaccg
     1621 ccctgccccc cctgtgcaga aagcccctgg tcctgatgag cggcagaggc agggattgcg
     1681 ccagctgcag ctccttcgcc agcgacgtgc tggaacacta ccgggccgag tgggagtaca
     1741 tccgggccca cgtgaccctg gtggacgccg gcgatcccat gaccgaggaa ctcgtgcgga
     1801 acccctaccc tctgtggtac gatcccgccg aggccggacc ctggatgacc ctgaccaagg
     1861 tgtccgaagt gcgcagggct gagggggatg tggtggccgc tgccagcggc gcctcttcta
     1921 catctggcgc ccctagccac gacgtggccc agaccctgca gcagtccgac gtggaagatt
     1981 acaccctgtt caacatgggc gtgcggcgga aggccgcatt tctgctgcag tccccaaccg
     2041 tgcaggccac ccaccactgg tcccggctgg tcaagagagc cttcgccggc cagagcgagt
     2101 acgtgctgcg gatcgtgttc gcctaccccc acaacggcag cgtgatgcgg gacatcgtga
     2161 acgaggaagt gaccgtcggc ggcagccagg ccgactacct gcacttcttc aacaaggccc
     2221 agccattctt ccactgcgtg aagaaagccg tgcgcgtgat ggactggctg cagagatacg
     2281 gcgactatgg cgcgcccggc agcctgcacc acatcctgga cgcccagaag atgctgtgga
     2341 accacaggga caggaacctg cctcctctgg cccctctggg cccccaccac catcaccacc
     2401 attgagtcta gacacgtgat taattaagga tcccccgacc tcgacctctg gctaataaag
     2461 gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac
     2521 atatgggagg gcaaatcatt tggtcgagat ccctcggaga tctctagcta gaggatcgat
     2581 ccccgccccg gacgaactaa acctgactac gacatctctg ccccttcttc gcggggcagt
     2641 gcatgtaatc ccttcagttg gttggtacaa cttgccaact gggccctgtt ccacatgtga
     2701 cacggggggg gaccaaacac aaaggggttc tctgactgta gttgacatcc ttataaatgg
     2761 atgtgcacat ttgccaacac tgagtggctt tcatcctgga gcagactttg cagtctgtgg
     2821 actgcaacac aacattgcct ttatgtgtaa ctcttggctg aagctcttac accaatgctg
     2881 ggggacatgt acctcccagg ggcccaggaa gactacggga ggctacacca acgtcaatca
     2941 gaggggcctg tgtagctacc gataagcgga ccctcaagag ggcattagca atagtgttta
     3001 taaggccccc ttgttaaccc taaacgggta gcatatgctt cccgggtagt agtatatact
     3061 atccagacta accctaattc aatagcatat gttacccaac gggaagcata tgctatcgaa
     3121 ttagggttag taaaagggtc ctaaggaaca gcgatatctc ccaccccatg agctgtcacg
     3181 gttttattta catggggtca ggattccacg agggtagtga accattttag tcacaagggc
     3241 agtggctgaa gatcaaggag cgggcagtga actctcctga atcttcgcct gcttcttcat
     3301 tctccttcgt ttagctaata gaataactgc tgagttgtga acagtaaggt gtatgtgagg
     3361 tgctcgaaaa caaggtttca ggtgacgccc ccagaataaa atttggacgg ggggttcagt
     3421 ggtggcattg tgctatgaca ccaatataac cctcacaaac cccttgggca ataaatacta
     3481 gtgtaggaat gaaacattct gaatatcttt aacaatagaa atccatgggg tggggacaag
     3541 ccgtaaagac tggatgtcca tctcacacga atttatggct atgggcaaca cataatccta
     3601 gtgcaatatg atactggggt tattaagatg tgtcccaggc agggaccaag acaggtgaac
     3661 catgttgtta cactctattt gtaacaaggg gaaagagagt ggacgccgac agcagcggac
     3721 tccactggtt gtctctaaca cccccgaaaa ttaaacgggg ctccacgcca atggggccca
     3781 taaacaaaga caagtggcca ctcttttttt tgaaattgtg gagtgggggc acgcgtcagc
     3841 ccccacacgc cgccctgcgg ttttggactg taaaataagg gtgtaataac ttggctgatt
     3901 gtaaccccgc taaccactgc ggtcaaacca cttgcccaca aaaccactaa tggcaccccg
     3961 gggaatacct gcataagtag gtgggcgggc caagataggg gcgcgattgc tgcgatctgg
     4021 aggacaaatt acacacactt gcgcctgagc gccaagcaca gggttgttgg tcctcatatt
     4081 cacgaggtcg ctgagagcac ggtgggctaa tgttgccatg ggtagcatat actacccaaa
     4141 tatctggata gcatatgcta tcctaatcta tatctgggta gcataggcta tcctaatcta
     4201 tatctgggta gcatatgcta tcctaatcta tatctgggta gtatatgcta tcctaattta
     4261 tatctgggta gcataggcta tcctaatcta tatctgggta gcatatgcta tcctaatcta
     4321 tatctgggta gtatatgcta tcctaatctg tatccgggta gcatatgcta tcctaataga
     4381 gattagggta gtatatgcta tcctaattta tatctgggta gcatatacta cccaaatatc
     4441 tggatagcat atgctatcct aatctatatc tgggtagcat atgctatcct aatctatatc
     4501 tgggtagcat aggctatcct aatctatatc tgggtagcat atgctatcct aatctatatc
     4561 tgggtagtat atgctatcct aatttatatc tgggtagcat aggctatcct aatctatatc
     4621 tgggtagcat atgctatcct aatctatatc tgggtagtat atgctatcct aatctgtatc
     4681 cgggtagcat atgctatcct catgataagc tgtcaaacat gagaattttc ttgaagacga
     4741 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     4801 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     4861 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     4921 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     4981 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     5041 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     5101 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     5161 ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat
     5221 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     5281 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     5341 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     5401 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     5461 cgtgacacca cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa
     5521 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     5581 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     5641 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     5701 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     5761 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     5821 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     5881 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5941 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     6001 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     6061 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     6121 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     6181 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     6241 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     6301 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     6361 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     6421 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     6481 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     6541 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     6601 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     6661 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     6721 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     6781 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     6841 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
     6901 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
     6961 gattacgcca agctctagct agaggtcgac caattctcat gtttgacagc ttatcatcgc
     7021 agatccgggc aacgttgttg ccattgctgc aggcgcagaa ctggtaggta tggaagatct
     7081 atacattgaa tcaatattgg caattagcca tattagtcat tggttatata gcataaatca
     7141 atattggcta ttggccattg catacgttgt atctatatca taatatgtac atttatattg
     7201 gctcatgtcc aatatgaccg c
//
