LOCUS       sequence-262328-       10104 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 10104)
  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 10104)
  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: D35_Ld_292300
FEATURES             Location/Qualifiers
     source          1..10104
                     /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             5268..5285
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    5337..5392
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5391..5410)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(7628..7646)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7714..7818
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7819..7887,7888..8679)
                     /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(8037..8056)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8850..9438
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9339..9358
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9592..9609
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9726..9747
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9762..9767,9768..9785,9786..9792)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9800..9816
                     /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     9805..9827
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9824..9840
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9824..9840
                     /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 ccctgggagc cgattctgcc acccccctgc tgtgcgacaa cgacgccaca ctgtacacca
     1261 ccggccctga tggcaccgcc ctgggcatcc tgctgtacga gggcgtggtg ccctgcgcca
     1321 gcaacgagac aatcttcatc acaatcgcca aggaccccag cccccccaac gccgtggtgg
     1381 ctctgagcga gtggtccttc ctggtgtcca tcaacaacag agcccctgcc accttcaccc
     1441 tgtacgccat gtgcggcaac accatcaagt gccagagcac cctgacctac gccctgagcc
     1501 agaccatggc cagcagcagc tccagcgacg tgctgcccct gagcagccct agctctagca
     1561 gcgccgactc tgtgctgcct aacaccacct accccctgtg ctcccacgcc tccaaggcca
     1621 gatcctacag cgtgggcgag gtgtaccacg acaacttccc catggacatc caggacggca
     1681 gctgcgtgta cacaaccagc atcaccggcg agtccaccgt gggcacagtg gtggtggcca
     1741 cctccggcct gagctacacc ttcagctcct ccaagacagc cgccgtgggc tacaacattc
     1801 tgctgggcta tatcgtgcgg tgcaacggca tcacagtgtg cgccgacacc gtggatgtgc
     1861 tgctggaaag cgccagcgtg accaccgccc ctccaacaag cacacccgcc cctacccctg
     1921 cccctcccga gacactgcct tggtgcagcc accggacata cgccaacacc taccggatcg
     1981 gcgaagtgta cttcggcgac ctgctgctcg acgtgaagga caccgagtgc cggtacaccg
     2041 ccaccatcga ggaccctagc gtgggaacag ccgtggtcgc cgagcctagc gaccccctgg
     2101 gctacgtgta cagcacctcc gacgataccg tgccccagaa cacctccatc ccttacacca
     2161 ttgcctgcaa tggcgtgtcc atttgccacg gcctggtggt catccacctg acttacgacc
     2221 ctcccgcctg ccccaacagc gtcgtgacct atctgctcga gcctggcggc tccgtgaccg
     2281 gcgccatcag cagcggaaca gtgtgtagca gcggcgccac acctagcccc agcatcctga
     2341 gccccgtgcc cggattcttc ctggacgcca gcggcgactt cttcttctac gccaccgacg
     2401 acgaggccga ggtcgtggcc atggtgctga tgaagtgcaa cgaagaggtg ctgtgccaga
     2461 tcaaagtgat gtttctggtg gcctacccca ccacagcccc acccaccacc tccaccacaa
     2521 ccaccgctgc ccccatcgtg atgtgcccca agctgttcag atacgaggtg cccaccgggc
     2581 agagcatcag cgataccatc gaccccgtga caacctccag ctgcaacttc accacatacc
     2641 tgctggtcaa ctccagcagc gacatcagag gatctctgct gctgaacctg ctcggcgagt
     2701 tctactacaa ggcccccaag agccaggccg tggactacgc cgctgtggac gtgtactgcg
     2761 ccaaggcctt cgtgtgccgg acccagctgg tgttcgtggc ctatacccct ctgccaacca
     2821 ccagcaccac catgcctcca ctgccccctt gcgccaatgt gtactactac cagaccagcc
     2881 ctggggtggc catcaatgcc tccctgaaca acatgcccgg ccaggacatg tgcgcccacg
     2941 gccggtactt taccctgaat acctccccac aggccggcag cgtgaacgtg tccctgatcg
     3001 gagacttcgt gtacctgcct cctaagtatg agggccagtt cagcttcatg ttcaccatgc
     3061 actgcatgaa ccagcagtac tgcaccggca ccgcttatct gctggtgtct aacgagtgga
     3121 ccctgaggcc agccccaacc gacagccctg tgaagcccga cagcaacgcc tctatcacca
     3181 acgagcagat cacctgtcac ggcacctgta acgcctccgc ctggaaaacc taccctaacc
     3241 ggaaagtgtg ggacatcacc cctggcgccg gatacgccag aaaggacggc cggaccgtga
     3301 atgacatgaa ggtcacatgg cggaacaaga gcctggtcgt gctggtctac agcctgattg
     3361 gcaacctggg cgtgcggttc cccaccttcg agcccatcac cacaagccag cgggcctaca
     3421 tggaacccaa ggacttcgcc ggaagcgtgg ccccaggcag ccccggcttt gagatgagct
     3481 gcctggccaa ccagggcaga accggcatgg gagaggacgt gtggaagtgg ttcagcctga
     3541 ccctggacag cggcacaggc ggcgtgggag cctactataa taccggccag agctggtatc
     3601 agaagttcgg cggcaagcac gtgaactgcg acatcttcac cgacccctgc agatacgccc
     3661 cactgctgac ccccgccaac tacaccaaca ccagcctggg ccggtggacc gtggacgtcg
     3721 acgactgtga tgccacctgg accggcgtgt tcgagcagag cagcatgggc aagatggtca
     3781 aatacgacgg ctcccccgtg tggtctttcg tgggcaagct ccagctccag ggcaccctgt
     3841 atagcgaggc cgtgcaggcc aggtcctggc tggaacctgg ccagtttgac gccctgtaca
     3901 gctcccacaa catcctgatc aacctgcacc agtttgtggc cgtgtccaag accggaatgc
     3961 aggaccctct gatcagcgtg gacgccgaat tcttcaccta cctggacgac gaaacccagg
     4021 accaggcctt tggcatcaac atgctcgtgt atcccttcgt ggacagctac atgaccacct
     4081 cttacgcccg ggacagacac gtgaaaggct tcaagtggat cctgcaggac tggatcagcc
     4141 caagcgacga gcactgccct acctgtaccg gcagcaaaat gaagtgcgtg gcacctctgc
     4201 agggcgtcga cgtgtcctac accggaccta gcttccccga gggcgactgt gccgatggcg
     4261 agggcagagt gcacctgttc aagggcccag ccacaatgcc cagcgactgc cccgactcca
     4321 agatgcatgt gttcaaccag accggcttct cccccacccg gaactgcaat accgcctacc
     4381 agaacatgac cctgcggggc atcgtgcctg gcagctctgg cctgaacgag actctgagcc
     4441 gggccttcca ctacgaggga accgtgcagc tggtcctgct gatggacgac cactcctaca
     4501 agcggctgga cctgcatctg tctatgtacg tgtccagact gagcaaggac agcccccgga
     4561 tgagaggcgg cctgtccacc tgtagaagcg gctcctactg gcccgtgctg gatcctctgg
     4621 gcagctccct ggcctccagc ccctttcctc tgtccgtggc tgccgtgaca cccctgtgta
     4681 tcgacgacct gtcctctagc tacggcccca acgactgggc cctgttcgcc gtgaacatgc
     4741 ctggcgtgaa gccaagcgaa gtgaccgtgc ggagcgtgta cgtgcagcac aacaactccc
     4801 ggatctacct gatctacaag gatcccagaa gcggagccgc ctccattccc aactccgacg
     4861 ccgacggcga ttggtggcag tacaagtacc cctttctggc cttccgggac ctggaagccc
     4921 acgtgaagaa caattccatc acctacagca cactgatgga aagctccgcc gctgccgccg
     4981 acgtggtgta cgccttcacc ttcatcccag gcgctctggg agtggatacc gacatcgagg
     5041 tggtggtcga agccctgatc cagcccagcg ccgcagccga gagcaccaca gagttccggc
     5101 gggtgctgca catcgatcca ctgctcacac agctgagcag acagggccct ccacctccct
     5161 atagcagctc tggcgcgccc ggcagcctgc accacatcct ggacgcccag aagatgctgt
     5221 ggaaccacag ggacaggaac ctgcctcctc tggcccctct gggcccccac caccatcacc
     5281 accattgagt ctagacacgt gattaattaa ggatcccccg acctcgacct ctggctaata
     5341 aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag
     5401 gacatatggg agggcaaatc atttggtcga gatccctcgg agatctctag ctagaggatc
     5461 gatccccgcc ccggacgaac taaacctgac tacgacatct ctgccccttc ttcgcggggc
     5521 agtgcatgta atcccttcag ttggttggta caacttgcca actgggccct gttccacatg
     5581 tgacacgggg ggggaccaaa cacaaagggg ttctctgact gtagttgaca tccttataaa
     5641 tggatgtgca catttgccaa cactgagtgg ctttcatcct ggagcagact ttgcagtctg
     5701 tggactgcaa cacaacattg cctttatgtg taactcttgg ctgaagctct tacaccaatg
     5761 ctgggggaca tgtacctccc aggggcccag gaagactacg ggaggctaca ccaacgtcaa
     5821 tcagaggggc ctgtgtagct accgataagc ggaccctcaa gagggcatta gcaatagtgt
     5881 ttataaggcc cccttgttaa ccctaaacgg gtagcatatg cttcccgggt agtagtatat
     5941 actatccaga ctaaccctaa ttcaatagca tatgttaccc aacgggaagc atatgctatc
     6001 gaattagggt tagtaaaagg gtcctaagga acagcgatat ctcccacccc atgagctgtc
     6061 acggttttat ttacatgggg tcaggattcc acgagggtag tgaaccattt tagtcacaag
     6121 ggcagtggct gaagatcaag gagcgggcag tgaactctcc tgaatcttcg cctgcttctt
     6181 cattctcctt cgtttagcta atagaataac tgctgagttg tgaacagtaa ggtgtatgtg
     6241 aggtgctcga aaacaaggtt tcaggtgacg cccccagaat aaaatttgga cggggggttc
     6301 agtggtggca ttgtgctatg acaccaatat aaccctcaca aaccccttgg gcaataaata
     6361 ctagtgtagg aatgaaacat tctgaatatc tttaacaata gaaatccatg gggtggggac
     6421 aagccgtaaa gactggatgt ccatctcaca cgaatttatg gctatgggca acacataatc
     6481 ctagtgcaat atgatactgg ggttattaag atgtgtccca ggcagggacc aagacaggtg
     6541 aaccatgttg ttacactcta tttgtaacaa ggggaaagag agtggacgcc gacagcagcg
     6601 gactccactg gttgtctcta acacccccga aaattaaacg gggctccacg ccaatggggc
     6661 ccataaacaa agacaagtgg ccactctttt ttttgaaatt gtggagtggg ggcacgcgtc
     6721 agcccccaca cgccgccctg cggttttgga ctgtaaaata agggtgtaat aacttggctg
     6781 attgtaaccc cgctaaccac tgcggtcaaa ccacttgccc acaaaaccac taatggcacc
     6841 ccggggaata cctgcataag taggtgggcg ggccaagata ggggcgcgat tgctgcgatc
     6901 tggaggacaa attacacaca cttgcgcctg agcgccaagc acagggttgt tggtcctcat
     6961 attcacgagg tcgctgagag cacggtgggc taatgttgcc atgggtagca tatactaccc
     7021 aaatatctgg atagcatatg ctatcctaat ctatatctgg gtagcatagg ctatcctaat
     7081 ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat
     7141 ttatatctgg gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat
     7201 ctatatctgg gtagtatatg ctatcctaat ctgtatccgg gtagcatatg ctatcctaat
     7261 agagattagg gtagtatatg ctatcctaat ttatatctgg gtagcatata ctacccaaat
     7321 atctggatag catatgctat cctaatctat atctgggtag catatgctat cctaatctat
     7381 atctgggtag cataggctat cctaatctat atctgggtag catatgctat cctaatctat
     7441 atctgggtag tatatgctat cctaatttat atctgggtag cataggctat cctaatctat
     7501 atctgggtag catatgctat cctaatctat atctgggtag tatatgctat cctaatctgt
     7561 atccgggtag catatgctat cctcatgata agctgtcaaa catgagaatt ttcttgaaga
     7621 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     7681 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     7741 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     7801 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     7861 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     7921 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     7981 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     8041 tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac
     8101 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     8161 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     8221 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     8281 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     8341 gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc
     8401 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     8461 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     8521 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     8581 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     8641 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     8701 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     8761 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     8821 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     8881 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     8941 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     9001 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     9061 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     9121 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     9181 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     9241 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     9301 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     9361 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     9421 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     9481 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     9541 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     9601 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     9661 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     9721 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     9781 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     9841 catgattacg ccaagctcta gctagaggtc gaccaattct catgtttgac agcttatcat
     9901 cgcagatccg ggcaacgttg ttgccattgc tgcaggcgca gaactggtag gtatggaaga
     9961 tctatacatt gaatcaatat tggcaattag ccatattagt cattggttat atagcataaa
    10021 tcaatattgg ctattggcca ttgcatacgt tgtatctata tcataatatg tacatttata
    10081 ttggctcatg tccaatatga ccgc
//
