LOCUS       sequence-262304-        6654 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 6654)
  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 6654)
  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: D9_Ld_190920
FEATURES             Location/Qualifiers
     source          1..6654
                     /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             1818..1835
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1887..1942
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1941..1960)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4178..4196)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4264..4368
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4369..4437,4438..5229)
                     /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(4587..4606)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5400..5988
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5889..5908
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6142..6159
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6276..6297
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6312..6317,6318..6335,6336..6342)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6350..6366
                     /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     6355..6377
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6374..6390
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6374..6390
                     /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 ccagctattg gagcgaggaa gtgaacagag tccggaccta cgccgccgtg aactacctgg
     1261 aacggatcgc caagcagccc aacgtgtccg ccctgcccag cggcctgctg ttcaccatcg
     1321 agcggagagg cttcggcgac agagcccctg ccgccgagga caagtgcgag atgcactaca
     1381 ccatccacta ccggttcccc ggcatcgtgg aaagcacccg gcaccacccc taccccgtgc
     1441 ggagaagccc tagccagctg atccctggca tggccgaggc catgcagctg atgcgcgagg
     1501 gcgaccggtg gtatctgcac gtgccctacc agctgggcta cggcaaagag ggctgcaaag
     1561 aaaagaaggt gcccagcctg agcaacctgc gggtggaaat cgagctgtat aagtgcgaga
     1621 gcgccagcgg caagaccagc gccgagatcg acgcctacct ggccaagtac atgaagaccc
     1681 ggatccccga gaaggccgct cccgtggact acaccgacct gggcgcgccc ggcagcctgc
     1741 accacatcct ggacgcccag aagatgctgt ggaaccacag ggacaggaac ctgcctcctc
     1801 tggcccctct gggcccccac caccatcacc accattgagt ctagacacgt gattaattaa
     1861 ggatcccccg acctcgacct ctggctaata aaggaaattt attttcattg caatagtgtg
     1921 ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc atttggtcga
     1981 gatccctcgg agatctctag ctagaggatc gatccccgcc ccggacgaac taaacctgac
     2041 tacgacatct ctgccccttc ttcgcggggc agtgcatgta atcccttcag ttggttggta
     2101 caacttgcca actgggccct gttccacatg tgacacgggg ggggaccaaa cacaaagggg
     2161 ttctctgact gtagttgaca tccttataaa tggatgtgca catttgccaa cactgagtgg
     2221 ctttcatcct ggagcagact ttgcagtctg tggactgcaa cacaacattg cctttatgtg
     2281 taactcttgg ctgaagctct tacaccaatg ctgggggaca tgtacctccc aggggcccag
     2341 gaagactacg ggaggctaca ccaacgtcaa tcagaggggc ctgtgtagct accgataagc
     2401 ggaccctcaa gagggcatta gcaatagtgt ttataaggcc cccttgttaa ccctaaacgg
     2461 gtagcatatg cttcccgggt agtagtatat actatccaga ctaaccctaa ttcaatagca
     2521 tatgttaccc aacgggaagc atatgctatc gaattagggt tagtaaaagg gtcctaagga
     2581 acagcgatat ctcccacccc atgagctgtc acggttttat ttacatgggg tcaggattcc
     2641 acgagggtag tgaaccattt tagtcacaag ggcagtggct gaagatcaag gagcgggcag
     2701 tgaactctcc tgaatcttcg cctgcttctt cattctcctt cgtttagcta atagaataac
     2761 tgctgagttg tgaacagtaa ggtgtatgtg aggtgctcga aaacaaggtt tcaggtgacg
     2821 cccccagaat aaaatttgga cggggggttc agtggtggca ttgtgctatg acaccaatat
     2881 aaccctcaca aaccccttgg gcaataaata ctagtgtagg aatgaaacat tctgaatatc
     2941 tttaacaata gaaatccatg gggtggggac aagccgtaaa gactggatgt ccatctcaca
     3001 cgaatttatg gctatgggca acacataatc ctagtgcaat atgatactgg ggttattaag
     3061 atgtgtccca ggcagggacc aagacaggtg aaccatgttg ttacactcta tttgtaacaa
     3121 ggggaaagag agtggacgcc gacagcagcg gactccactg gttgtctcta acacccccga
     3181 aaattaaacg gggctccacg ccaatggggc ccataaacaa agacaagtgg ccactctttt
     3241 ttttgaaatt gtggagtggg ggcacgcgtc agcccccaca cgccgccctg cggttttgga
     3301 ctgtaaaata agggtgtaat aacttggctg attgtaaccc cgctaaccac tgcggtcaaa
     3361 ccacttgccc acaaaaccac taatggcacc ccggggaata cctgcataag taggtgggcg
     3421 ggccaagata ggggcgcgat tgctgcgatc tggaggacaa attacacaca cttgcgcctg
     3481 agcgccaagc acagggttgt tggtcctcat attcacgagg tcgctgagag cacggtgggc
     3541 taatgttgcc atgggtagca tatactaccc aaatatctgg atagcatatg ctatcctaat
     3601 ctatatctgg gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat
     3661 ctatatctgg gtagtatatg ctatcctaat ttatatctgg gtagcatagg ctatcctaat
     3721 ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat
     3781 ctgtatccgg gtagcatatg ctatcctaat agagattagg gtagtatatg ctatcctaat
     3841 ttatatctgg gtagcatata ctacccaaat atctggatag catatgctat cctaatctat
     3901 atctgggtag catatgctat cctaatctat atctgggtag cataggctat cctaatctat
     3961 atctgggtag catatgctat cctaatctat atctgggtag tatatgctat cctaatttat
     4021 atctgggtag cataggctat cctaatctat atctgggtag catatgctat cctaatctat
     4081 atctgggtag tatatgctat cctaatctgt atccgggtag catatgctat cctcatgata
     4141 agctgtcaaa catgagaatt ttcttgaaga cgaaagggcc tcgtgatacg cctattttta
     4201 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     4261 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     4321 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     4381 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     4441 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     4501 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     4561 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc
     4621 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     4681 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     4741 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     4801 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     4861 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg
     4921 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     4981 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     5041 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     5101 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     5161 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     5221 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     5281 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     5341 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     5401 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     5461 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     5521 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     5581 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     5641 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     5701 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     5761 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     5821 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     5881 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     5941 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     6001 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     6061 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     6121 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     6181 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     6241 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     6301 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     6361 taacaatttc acacaggaaa cagctatgac catgattacg ccaagctcta gctagaggtc
     6421 gaccaattct catgtttgac agcttatcat cgcagatccg ggcaacgttg ttgccattgc
     6481 tgcaggcgca gaactggtag gtatggaaga tctatacatt gaatcaatat tggcaattag
     6541 ccatattagt cattggttat atagcataaa tcaatattgg ctattggcca ttgcatacgt
     6601 tgtatctata tcataatatg tacatttata ttggctcatg tccaatatga ccgc
//
