LOCUS       sequence-262348-        6618 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 6618)
  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 6618)
  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: D56_Ld_141480
FEATURES             Location/Qualifiers
     source          1..6618
                     /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             1782..1799
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1851..1906
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1905..1924)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4142..4160)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4228..4332
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4333..4401,4402..5193)
                     /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(4551..4570)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5364..5952
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5853..5872
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6106..6123
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6240..6261
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6276..6281,6282..6299,6300..6306)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6314..6330
                     /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     6319..6341
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6338..6354
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6338..6354
                     /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 gcggccgccg
     1201 tgctgcctgc tacagccctg agagtgaagc ccaccatcac cgccctgacc ttcgacgtgg
     1261 gcaaagagga agtctgcctg tacagcatgg gcaaggactt ctacgagggc gtgaccatgc
     1321 actaccacgt gatggaaggc gagaacgact tcgacgtgtt catccgggac gtggacggcc
     1381 acgtgatcta cgccagctat gccggcgagc acggcgccga ggaccgggtg tactttacca
     1441 cccacgccaa caaagagcac gcctactgca tcgacaaccg ggactacagc ggcgagctga
     1501 aagtgatcaa gatggaagtg ggcctgacca gcctgaagcg gtggaagaac cggatcgacc
     1561 ccctgcggaa gctgatgacc cggaccgacg gcttcgtgat gggcatgcac gacgaccaga
     1621 tcctgctgcg gatgaaggaa cagggcatcc gcgagtgggt ggaaaagatc ttcaccatgc
     1681 tgacaggcgc gcccggcagc ctgcaccaca tcctggacgc ccagaagatg ctgtggaacc
     1741 acagggacag gaacctgcct cctctggccc ctctgggccc ccaccaccat caccaccatt
     1801 gagtctagac acgtgattaa ttaaggatcc cccgacctcg acctctggct aataaaggaa
     1861 atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata
     1921 tgggagggca aatcatttgg tcgagatccc tcggagatct ctagctagag gatcgatccc
     1981 cgccccggac gaactaaacc tgactacgac atctctgccc cttcttcgcg gggcagtgca
     2041 tgtaatccct tcagttggtt ggtacaactt gccaactggg ccctgttcca catgtgacac
     2101 ggggggggac caaacacaaa ggggttctct gactgtagtt gacatcctta taaatggatg
     2161 tgcacatttg ccaacactga gtggctttca tcctggagca gactttgcag tctgtggact
     2221 gcaacacaac attgccttta tgtgtaactc ttggctgaag ctcttacacc aatgctgggg
     2281 gacatgtacc tcccaggggc ccaggaagac tacgggaggc tacaccaacg tcaatcagag
     2341 gggcctgtgt agctaccgat aagcggaccc tcaagagggc attagcaata gtgtttataa
     2401 ggcccccttg ttaaccctaa acgggtagca tatgcttccc gggtagtagt atatactatc
     2461 cagactaacc ctaattcaat agcatatgtt acccaacggg aagcatatgc tatcgaatta
     2521 gggttagtaa aagggtccta aggaacagcg atatctccca ccccatgagc tgtcacggtt
     2581 ttatttacat ggggtcagga ttccacgagg gtagtgaacc attttagtca caagggcagt
     2641 ggctgaagat caaggagcgg gcagtgaact ctcctgaatc ttcgcctgct tcttcattct
     2701 ccttcgttta gctaatagaa taactgctga gttgtgaaca gtaaggtgta tgtgaggtgc
     2761 tcgaaaacaa ggtttcaggt gacgccccca gaataaaatt tggacggggg gttcagtggt
     2821 ggcattgtgc tatgacacca atataaccct cacaaacccc ttgggcaata aatactagtg
     2881 taggaatgaa acattctgaa tatctttaac aatagaaatc catggggtgg ggacaagccg
     2941 taaagactgg atgtccatct cacacgaatt tatggctatg ggcaacacat aatcctagtg
     3001 caatatgata ctggggttat taagatgtgt cccaggcagg gaccaagaca ggtgaaccat
     3061 gttgttacac tctatttgta acaaggggaa agagagtgga cgccgacagc agcggactcc
     3121 actggttgtc tctaacaccc ccgaaaatta aacggggctc cacgccaatg gggcccataa
     3181 acaaagacaa gtggccactc ttttttttga aattgtggag tgggggcacg cgtcagcccc
     3241 cacacgccgc cctgcggttt tggactgtaa aataagggtg taataacttg gctgattgta
     3301 accccgctaa ccactgcggt caaaccactt gcccacaaaa ccactaatgg caccccgggg
     3361 aatacctgca taagtaggtg ggcgggccaa gataggggcg cgattgctgc gatctggagg
     3421 acaaattaca cacacttgcg cctgagcgcc aagcacaggg ttgttggtcc tcatattcac
     3481 gaggtcgctg agagcacggt gggctaatgt tgccatgggt agcatatact acccaaatat
     3541 ctggatagca tatgctatcc taatctatat ctgggtagca taggctatcc taatctatat
     3601 ctgggtagca tatgctatcc taatctatat ctgggtagta tatgctatcc taatttatat
     3661 ctgggtagca taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat
     3721 ctgggtagta tatgctatcc taatctgtat ccgggtagca tatgctatcc taatagagat
     3781 tagggtagta tatgctatcc taatttatat ctgggtagca tatactaccc aaatatctgg
     3841 atagcatatg ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg
     3901 gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg
     3961 gtagtatatg ctatcctaat ttatatctgg gtagcatagg ctatcctaat ctatatctgg
     4021 gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat ctgtatccgg
     4081 gtagcatatg ctatcctcat gataagctgt caaacatgag aattttcttg aagacgaaag
     4141 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     4201 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     4261 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     4321 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     4381 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     4441 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     4501 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     4561 gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     4621 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     4681 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     4741 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     4801 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     4861 gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     4921 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     4981 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     5041 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     5101 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     5161 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     5221 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     5281 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     5341 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     5401 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     5461 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     5521 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     5581 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     5641 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     5701 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     5761 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     5821 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     5881 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     5941 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     6001 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     6061 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     6121 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     6181 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     6241 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     6301 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     6361 tacgccaagc tctagctaga ggtcgaccaa ttctcatgtt tgacagctta tcatcgcaga
     6421 tccgggcaac gttgttgcca ttgctgcagg cgcagaactg gtaggtatgg aagatctata
     6481 cattgaatca atattggcaa ttagccatat tagtcattgg ttatatagca taaatcaata
     6541 ttggctattg gccattgcat acgttgtatc tatatcataa tatgtacatt tatattggct
     6601 catgtccaat atgaccgc
//
