LOCUS       sequence-262300-        7704 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 7704)
  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 7704)
  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: D5_Ld_280600
FEATURES             Location/Qualifiers
     source          1..7704
                     /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"
     regulatory      2112..2121
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2868..2885
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2937..2992
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2991..3010)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5228..5246)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5314..5418
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5419..5487,5488..6279)
                     /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(5637..5656)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6450..7038
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6939..6958
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7192..7209
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7326..7347
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7362..7367,7368..7385,7386..7392)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7400..7416
                     /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     7405..7427
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7424..7440
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7424..7440
                     /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 ccgctcaggg cgatcctgag agagccgaca gcgaggaacc cagatgcggc ttcgacgagc
     1261 tggaagcccg gatgatcggc acccgggtgt ccgtgatcag ccgggtggaa cctcctaccg
     1321 gcgaactggc cgtggccgct gctgctacag gcgcctggca gcccatccgg atcgccgtgt
     1381 tcaccgagga catcagcaac agcagccagc actgcaccgc cagcggccag agcagaccca
     1441 ccttcagagg cggcagagtg acctgctctg ccgccgacgt gctgacccgc gagaagaaac
     1501 gggtgctgct ggaactgctg atccccagcg ccgtgcagct gcatcaggaa cggctgaacg
     1561 tccagcgcga gaacggcaac atcgtggtgt ccccattcat caagaagcac agcatctgcg
     1621 gccagttcag catccccgag gaacacatga agaccggcgt gcccgacgcc gacttcgtgc
     1681 tgtacatgag cgccgctccc accagcggct ctgtgatcgc ctgggccgtg aagtgccaga
     1741 gcttcgacaa cggccggtcc agcgtgggcg tggccaccat cagccccaag tacatcaccg
     1801 ccgagcccaa gaccgtgcgg gtggtggccc atgaagtgct gcacgccctg ggcttcacca
     1861 gaagcgtgtt caagcagcag aacatgctgg tcatggccag cttccggggc aagagcccca
     1921 gccccgtgat cagatccgcc aacgtggtgg ctcaggccca gctgcactac ggctgcaaga
     1981 cccaggccag catggaactg gaagatgagg gcggcaaggg caccgtgtcc agccactgga
     2041 agcggcggag cgccaaggat gagctgatgg ccggcttcag cggcgtggga gtgtacagcg
     2101 ccctgacaat cgccgccatg gaagataccg gctactacca gggcaactac gccaaggccg
     2161 agcccatggc ctacggccac gaagtgggct gcaagctgag cagcgagaga tgcgtgatca
     2221 agagcaccag ccagatcccc gccatgttct gcgacgcccc cgatgcccct tggagctgca
     2281 ccagcgatcg gagaggcatc ggccggtgca tcctgaccag ctacaacagc aacctgccca
     2341 cctacttcca gtacttcggc gaccccagac tgggcggacc cgaccccctg atggacttct
     2401 gccctttcgt gcgggctgcc gacgacacca tgtgcgccgc caagaccaac gccctgaagg
     2461 gcagcgtgta cggcgtgatg agcagatgcg tggacacccc tgccggcttc tccatcgacg
     2521 actctgccgt gcagcagcac ggaatctgcg ccgaggtgca gtgcggcagc agcgcctacg
     2581 gcgtgaagat caacggcgcc agcgccttca gagactgccc ccctggcagc acctacaacc
     2641 tgagcaccct gagccccagc ttcagcaagg gccacctcgt gtgccccagc tacgagagcg
     2701 tgtgcgccat caacatcaac gccagcctgt acgaagagta cagccggctg ctgaccgacc
     2761 acagcgtgac aggcgcgccc ggcagcctgc accacatcct ggacgcccag aagatgctgt
     2821 ggaaccacag ggacaggaac ctgcctcctc tggcccctct gggcccccac caccatcacc
     2881 accattgagt ctagacacgt gattaattaa ggatcccccg acctcgacct ctggctaata
     2941 aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag
     3001 gacatatggg agggcaaatc atttggtcga gatccctcgg agatctctag ctagaggatc
     3061 gatccccgcc ccggacgaac taaacctgac tacgacatct ctgccccttc ttcgcggggc
     3121 agtgcatgta atcccttcag ttggttggta caacttgcca actgggccct gttccacatg
     3181 tgacacgggg ggggaccaaa cacaaagggg ttctctgact gtagttgaca tccttataaa
     3241 tggatgtgca catttgccaa cactgagtgg ctttcatcct ggagcagact ttgcagtctg
     3301 tggactgcaa cacaacattg cctttatgtg taactcttgg ctgaagctct tacaccaatg
     3361 ctgggggaca tgtacctccc aggggcccag gaagactacg ggaggctaca ccaacgtcaa
     3421 tcagaggggc ctgtgtagct accgataagc ggaccctcaa gagggcatta gcaatagtgt
     3481 ttataaggcc cccttgttaa ccctaaacgg gtagcatatg cttcccgggt agtagtatat
     3541 actatccaga ctaaccctaa ttcaatagca tatgttaccc aacgggaagc atatgctatc
     3601 gaattagggt tagtaaaagg gtcctaagga acagcgatat ctcccacccc atgagctgtc
     3661 acggttttat ttacatgggg tcaggattcc acgagggtag tgaaccattt tagtcacaag
     3721 ggcagtggct gaagatcaag gagcgggcag tgaactctcc tgaatcttcg cctgcttctt
     3781 cattctcctt cgtttagcta atagaataac tgctgagttg tgaacagtaa ggtgtatgtg
     3841 aggtgctcga aaacaaggtt tcaggtgacg cccccagaat aaaatttgga cggggggttc
     3901 agtggtggca ttgtgctatg acaccaatat aaccctcaca aaccccttgg gcaataaata
     3961 ctagtgtagg aatgaaacat tctgaatatc tttaacaata gaaatccatg gggtggggac
     4021 aagccgtaaa gactggatgt ccatctcaca cgaatttatg gctatgggca acacataatc
     4081 ctagtgcaat atgatactgg ggttattaag atgtgtccca ggcagggacc aagacaggtg
     4141 aaccatgttg ttacactcta tttgtaacaa ggggaaagag agtggacgcc gacagcagcg
     4201 gactccactg gttgtctcta acacccccga aaattaaacg gggctccacg ccaatggggc
     4261 ccataaacaa agacaagtgg ccactctttt ttttgaaatt gtggagtggg ggcacgcgtc
     4321 agcccccaca cgccgccctg cggttttgga ctgtaaaata agggtgtaat aacttggctg
     4381 attgtaaccc cgctaaccac tgcggtcaaa ccacttgccc acaaaaccac taatggcacc
     4441 ccggggaata cctgcataag taggtgggcg ggccaagata ggggcgcgat tgctgcgatc
     4501 tggaggacaa attacacaca cttgcgcctg agcgccaagc acagggttgt tggtcctcat
     4561 attcacgagg tcgctgagag cacggtgggc taatgttgcc atgggtagca tatactaccc
     4621 aaatatctgg atagcatatg ctatcctaat ctatatctgg gtagcatagg ctatcctaat
     4681 ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat
     4741 ttatatctgg gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat
     4801 ctatatctgg gtagtatatg ctatcctaat ctgtatccgg gtagcatatg ctatcctaat
     4861 agagattagg gtagtatatg ctatcctaat ttatatctgg gtagcatata ctacccaaat
     4921 atctggatag catatgctat cctaatctat atctgggtag catatgctat cctaatctat
     4981 atctgggtag cataggctat cctaatctat atctgggtag catatgctat cctaatctat
     5041 atctgggtag tatatgctat cctaatttat atctgggtag cataggctat cctaatctat
     5101 atctgggtag catatgctat cctaatctat atctgggtag tatatgctat cctaatctgt
     5161 atccgggtag catatgctat cctcatgata agctgtcaaa catgagaatt ttcttgaaga
     5221 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     5281 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     5341 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     5401 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     5461 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     5521 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     5581 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     5641 tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac
     5701 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     5761 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     5821 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     5881 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     5941 gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc
     6001 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     6061 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     6121 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     6181 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     6241 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     6301 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     6361 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     6421 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     6481 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     6541 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     6601 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     6661 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     6721 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     6781 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     6841 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     6901 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     6961 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     7021 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     7081 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     7141 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     7201 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     7261 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     7321 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     7381 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     7441 catgattacg ccaagctcta gctagaggtc gaccaattct catgtttgac agcttatcat
     7501 cgcagatccg ggcaacgttg ttgccattgc tgcaggcgca gaactggtag gtatggaaga
     7561 tctatacatt gaatcaatat tggcaattag ccatattagt cattggttat atagcataaa
     7621 tcaatattgg ctattggcca ttgcatacgt tgtatctata tcataatatg tacatttata
     7681 ttggctcatg tccaatatga ccgc
//
