LOCUS       sequence-274893-        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"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     201..225
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     706..725
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      748..757
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1285..1302
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1354..1409
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1408..1427)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3645..3663)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3731..3835
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3836..3904,3905..4696)
                     /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(4054..4073)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4867..5455
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5356..5375
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5609..5626
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5743..5764
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5779..5784,5785..5802,5803..5809)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5817..5833
                     /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     5822..5844
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5841..5857
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5841..5857
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        6128..6507
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta caccaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcctcac tctcttccgc
      241 atcgctgtct gcgagggcca gctgttgggc tcgcggttga ggacaaactc ttcgcggtct
      301 ttccagtact cttggatcgg aaacccgtcg gcctccgaac ggtactccgc caccgaggga
      361 cctgagcgag tccgcatcga ccggatcgga aaacctctcg agaaaggcgt ctaaccagtc
      421 acagtcgcaa ggtaggctga gcaccgtggc gggcggcagc gggtggcggt cggggttgtt
      481 tctggcggag gtgctgctga tgatgtaatt aaagtaggcg gtcttgagac ggcggatggt
      541 cgaggtgagg tgtggcaggc ttgagatcca gctgttgggg tgagtactcc ctctcaaaag
      601 cgggcattac ttctgcgcta agattgtcag tttccaaaaa cgaggaggat ttgatattca
      661 cctggcccga tctggccata cacttgagtg acaatgacat ccactttgcc tttctctcca
      721 caggtgtcca ctcccaggtc caagtttgcc accatggagt ttcagaccca ggtactcatg
      781 tccctgctgc tctgcatgtc tggtgcggcc gccaccgcct cttctacccc tgcccctacc
      841 gacctgatca ccgtggccgg aagagtgcac gtgcggaacg aggtggtgca gtacgtggtc
      901 gtgcgggtgg tggacgacaa gaccggggtg gtcgtcagaa gcgcccctct ggacgccacc
      961 agaagcgtgt ccttcgccaa tatcgccagc agcgccgtgt ccctggaagc caccgtggaa
     1021 ctgcccgacc acctgttcga gctggatgcc agctccagcg tgctgaagtc cgccatcagc
     1081 accagcagct ccaccacccc cgtgcagctg accatctcca caatcagcaa ggccgagggc
     1141 gctgccagaa aggcccctac acctacagcc ggcagcagcg gcgcttctgg cgcgcccggc
     1201 agcctgcacc acatcctgga cgcccagaag atgctgtgga accacaggga caggaacctg
     1261 cctcctctgg cccctctggg cccccaccac catcaccacc attgagtcta gacacgtgat
     1321 taattaagga tcccccgacc tcgacctctg gctaataaag gaaatttatt ttcattgcaa
     1381 tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt
     1441 tggtcgagat ccctcggaga tctctagcta gaggatcgat ccccgccccg gacgaactaa
     1501 acctgactac gacatctctg ccccttcttc gcggggcagt gcatgtaatc ccttcagttg
     1561 gttggtacaa cttgccaact gggccctgtt ccacatgtga cacggggggg gaccaaacac
     1621 aaaggggttc tctgactgta gttgacatcc ttataaatgg atgtgcacat ttgccaacac
     1681 tgagtggctt tcatcctgga gcagactttg cagtctgtgg actgcaacac aacattgcct
     1741 ttatgtgtaa ctcttggctg aagctcttac accaatgctg ggggacatgt acctcccagg
     1801 ggcccaggaa gactacggga ggctacacca acgtcaatca gaggggcctg tgtagctacc
     1861 gataagcgga ccctcaagag ggcattagca atagtgttta taaggccccc ttgttaaccc
     1921 taaacgggta gcatatgctt cccgggtagt agtatatact atccagacta accctaattc
     1981 aatagcatat gttacccaac gggaagcata tgctatcgaa ttagggttag taaaagggtc
     2041 ctaaggaaca gcgatatctc ccaccccatg agctgtcacg gttttattta catggggtca
     2101 ggattccacg agggtagtga accattttag tcacaagggc agtggctgaa gatcaaggag
     2161 cgggcagtga actctcctga atcttcgcct gcttcttcat tctccttcgt ttagctaata
     2221 gaataactgc tgagttgtga acagtaaggt gtatgtgagg tgctcgaaaa caaggtttca
     2281 ggtgacgccc ccagaataaa atttggacgg ggggttcagt ggtggcattg tgctatgaca
     2341 ccaatataac cctcacaaac cccttgggca ataaatacta gtgtaggaat gaaacattct
     2401 gaatatcttt aacaatagaa atccatgggg tggggacaag ccgtaaagac tggatgtcca
     2461 tctcacacga atttatggct atgggcaaca cataatccta gtgcaatatg atactggggt
     2521 tattaagatg tgtcccaggc agggaccaag acaggtgaac catgttgtta cactctattt
     2581 gtaacaaggg gaaagagagt ggacgccgac agcagcggac tccactggtt gtctctaaca
     2641 cccccgaaaa ttaaacgggg ctccacgcca atggggccca taaacaaaga caagtggcca
     2701 ctcttttttt tgaaattgtg gagtgggggc acgcgtcagc ccccacacgc cgccctgcgg
     2761 ttttggactg taaaataagg gtgtaataac ttggctgatt gtaaccccgc taaccactgc
     2821 ggtcaaacca cttgcccaca aaaccactaa tggcaccccg gggaatacct gcataagtag
     2881 gtgggcgggc caagataggg gcgcgattgc tgcgatctgg aggacaaatt acacacactt
     2941 gcgcctgagc gccaagcaca gggttgttgg tcctcatatt cacgaggtcg ctgagagcac
     3001 ggtgggctaa tgttgccatg ggtagcatat actacccaaa tatctggata gcatatgcta
     3061 tcctaatcta tatctgggta gcataggcta tcctaatcta tatctgggta gcatatgcta
     3121 tcctaatcta tatctgggta gtatatgcta tcctaattta tatctgggta gcataggcta
     3181 tcctaatcta tatctgggta gcatatgcta tcctaatcta tatctgggta gtatatgcta
     3241 tcctaatctg tatccgggta gcatatgcta tcctaataga gattagggta gtatatgcta
     3301 tcctaattta tatctgggta gcatatacta cccaaatatc tggatagcat atgctatcct
     3361 aatctatatc tgggtagcat atgctatcct aatctatatc tgggtagcat aggctatcct
     3421 aatctatatc tgggtagcat atgctatcct aatctatatc tgggtagtat atgctatcct
     3481 aatttatatc tgggtagcat aggctatcct aatctatatc tgggtagcat atgctatcct
     3541 aatctatatc tgggtagtat atgctatcct aatctgtatc cgggtagcat atgctatcct
     3601 catgataagc tgtcaaacat gagaattttc ttgaagacga aagggcctcg tgatacgcct
     3661 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     3721 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3781 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     3841 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     3901 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     3961 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     4021 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt
     4081 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     4141 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     4201 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     4261 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     4321 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc
     4381 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     4441 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     4501 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     4561 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     4621 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     4681 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     4741 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     4801 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     4861 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     4921 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     4981 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     5041 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5101 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5161 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5221 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5281 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5341 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5401 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5461 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     5521 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     5581 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     5641 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     5701 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     5761 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     5821 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agctctagct
     5881 agaggtcgac caattctcat gtttgacagc ttatcatcgc agatccgggc aacgttgttg
     5941 ccattgctgc aggcgcagaa ctggtaggta tggaagatct atacattgaa tcaatattgg
     6001 caattagcca tattagtcat tggttatata gcataaatca atattggcta ttggccattg
     6061 catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc aatatgaccg
     6121 ccatgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
     6181 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     6241 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     6301 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     6361 gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga cggtaaatgg
     6421 cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg gcagtacatc
     6481 tacgtattag tcatcgctat taccatg
//
