LOCUS       sequence-262305-        6744 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 6744)
  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 6744)
  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: D10_Ld_292250
FEATURES             Location/Qualifiers
     source          1..6744
                     /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             1908..1925
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1977..2032
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2031..2050)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4268..4286)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4354..4458
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4459..4527,4528..5319)
                     /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(4677..4696)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5490..6078
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5979..5998
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6232..6249
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6366..6387
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6402..6407,6408..6425,6426..6432)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6440..6456
                     /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     6445..6467
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6464..6480
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6464..6480
                     /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 ccttcttcgc ctacgtggac cagcccagcc actgggtgta cgaccctgtg cggctgcagc
     1261 agatcgccca gcagtctatc gccaccgccc acgccgccaa tggcggcaag gccacagcca
     1321 agcaggtcac agacgagaca atccggctga tcctggaaaa ctaccccaag accacccggt
     1381 acaccggcca ctggctgtgg aacaatgccg gcggagccat gggcagcatg accgtgctgc
     1441 actgcagctt cagcgagtac ctgatcatct tcggcagcag cgtgggcacc gagggccaca
     1501 ccggcagata cttttgggcc gacgacttct tcaacatcct cgtgggcgag cagtgggccg
     1561 ctctgcccgg cgtggaagag aaagaggtgt acagacccgg cgaccagcac atcctgccca
     1621 gaggcagcgc caagcagtac cggatgcccg aggaatgctg ggccctggaa tacgccagag
     1681 gcaacatcgt gtccatgctg ttcttcggct tcgccgacat gttcagcagc accctggacg
     1741 tggtcaccat ctaccagacc gtgatggaaa gcgccggcaa catgatccgg aacaccctga
     1801 tgggcaagat tggcgcgccc ggcagcctgc accacatcct ggacgcccag aagatgctgt
     1861 ggaaccacag ggacaggaac ctgcctcctc tggcccctct gggcccccac caccatcacc
     1921 accattgagt ctagacacgt gattaattaa ggatcccccg acctcgacct ctggctaata
     1981 aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag
     2041 gacatatggg agggcaaatc atttggtcga gatccctcgg agatctctag ctagaggatc
     2101 gatccccgcc ccggacgaac taaacctgac tacgacatct ctgccccttc ttcgcggggc
     2161 agtgcatgta atcccttcag ttggttggta caacttgcca actgggccct gttccacatg
     2221 tgacacgggg ggggaccaaa cacaaagggg ttctctgact gtagttgaca tccttataaa
     2281 tggatgtgca catttgccaa cactgagtgg ctttcatcct ggagcagact ttgcagtctg
     2341 tggactgcaa cacaacattg cctttatgtg taactcttgg ctgaagctct tacaccaatg
     2401 ctgggggaca tgtacctccc aggggcccag gaagactacg ggaggctaca ccaacgtcaa
     2461 tcagaggggc ctgtgtagct accgataagc ggaccctcaa gagggcatta gcaatagtgt
     2521 ttataaggcc cccttgttaa ccctaaacgg gtagcatatg cttcccgggt agtagtatat
     2581 actatccaga ctaaccctaa ttcaatagca tatgttaccc aacgggaagc atatgctatc
     2641 gaattagggt tagtaaaagg gtcctaagga acagcgatat ctcccacccc atgagctgtc
     2701 acggttttat ttacatgggg tcaggattcc acgagggtag tgaaccattt tagtcacaag
     2761 ggcagtggct gaagatcaag gagcgggcag tgaactctcc tgaatcttcg cctgcttctt
     2821 cattctcctt cgtttagcta atagaataac tgctgagttg tgaacagtaa ggtgtatgtg
     2881 aggtgctcga aaacaaggtt tcaggtgacg cccccagaat aaaatttgga cggggggttc
     2941 agtggtggca ttgtgctatg acaccaatat aaccctcaca aaccccttgg gcaataaata
     3001 ctagtgtagg aatgaaacat tctgaatatc tttaacaata gaaatccatg gggtggggac
     3061 aagccgtaaa gactggatgt ccatctcaca cgaatttatg gctatgggca acacataatc
     3121 ctagtgcaat atgatactgg ggttattaag atgtgtccca ggcagggacc aagacaggtg
     3181 aaccatgttg ttacactcta tttgtaacaa ggggaaagag agtggacgcc gacagcagcg
     3241 gactccactg gttgtctcta acacccccga aaattaaacg gggctccacg ccaatggggc
     3301 ccataaacaa agacaagtgg ccactctttt ttttgaaatt gtggagtggg ggcacgcgtc
     3361 agcccccaca cgccgccctg cggttttgga ctgtaaaata agggtgtaat aacttggctg
     3421 attgtaaccc cgctaaccac tgcggtcaaa ccacttgccc acaaaaccac taatggcacc
     3481 ccggggaata cctgcataag taggtgggcg ggccaagata ggggcgcgat tgctgcgatc
     3541 tggaggacaa attacacaca cttgcgcctg agcgccaagc acagggttgt tggtcctcat
     3601 attcacgagg tcgctgagag cacggtgggc taatgttgcc atgggtagca tatactaccc
     3661 aaatatctgg atagcatatg ctatcctaat ctatatctgg gtagcatagg ctatcctaat
     3721 ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat
     3781 ttatatctgg gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat
     3841 ctatatctgg gtagtatatg ctatcctaat ctgtatccgg gtagcatatg ctatcctaat
     3901 agagattagg gtagtatatg ctatcctaat ttatatctgg gtagcatata ctacccaaat
     3961 atctggatag catatgctat cctaatctat atctgggtag catatgctat cctaatctat
     4021 atctgggtag cataggctat cctaatctat atctgggtag catatgctat cctaatctat
     4081 atctgggtag tatatgctat cctaatttat atctgggtag cataggctat cctaatctat
     4141 atctgggtag catatgctat cctaatctat atctgggtag tatatgctat cctaatctgt
     4201 atccgggtag catatgctat cctcatgata agctgtcaaa catgagaatt ttcttgaaga
     4261 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     4321 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     4381 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4441 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     4501 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     4561 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     4621 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     4681 tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac
     4741 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     4801 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     4861 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     4921 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     4981 gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc
     5041 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     5101 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     5161 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     5221 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     5281 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     5341 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     5401 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5461 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5521 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5581 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     5641 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     5701 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     5761 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     5821 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     5881 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     5941 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     6001 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6061 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6121 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     6181 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     6241 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     6301 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     6361 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     6421 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     6481 catgattacg ccaagctcta gctagaggtc gaccaattct catgtttgac agcttatcat
     6541 cgcagatccg ggcaacgttg ttgccattgc tgcaggcgca gaactggtag gtatggaaga
     6601 tctatacatt gaatcaatat tggcaattag ccatattagt cattggttat atagcataaa
     6661 tcaatattgg ctattggcca ttgcatacgt tgtatctata tcataatatg tacatttata
     6721 ttggctcatg tccaatatga ccgc
//
