LOCUS       sequence-274542-        9324 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 9324)
  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 9324)
  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: D18_Ld_100590
FEATURES             Location/Qualifiers
     source          1..9324
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     1086..1105
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1128..1137
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4482..4499
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4551..4606
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4605..4624)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6842..6860)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6928..7032
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7033..7101,7102..7893)
                     /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(7251..7270)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8064..8652
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8553..8572
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8806..8823
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8940..8961
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8976..8981,8982..8999,9000..9006)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9014..9030
                     /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     9019..9041
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9038..9054
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9038..9054
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agatcctcac tctcttccgc atcgctgtct gcgagggcca gctgttgggc tcgcggttga
      661 ggacaaactc ttcgcggtct ttccagtact cttggatcgg aaacccgtcg gcctccgaac
      721 ggtactccgc caccgaggga cctgagcgag tccgcatcga ccggatcgga aaacctctcg
      781 agaaaggcgt ctaaccagtc acagtcgcaa ggtaggctga gcaccgtggc gggcggcagc
      841 gggtggcggt cggggttgtt tctggcggag gtgctgctga tgatgtaatt aaagtaggcg
      901 gtcttgagac ggcggatggt cgaggtgagg tgtggcaggc ttgagatcca gctgttgggg
      961 tgagtactcc ctctcaaaag cgggcattac ttctgcgcta agattgtcag tttccaaaaa
     1021 cgaggaggat ttgatattca cctggcccga tctggccata cacttgagtg acaatgacat
     1081 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caagtttgcc accatggagt
     1141 ttcagaccca ggtactcatg tccctgctgc tctgcatgtc tggtgcggcc gccaccgctg
     1201 agggaaagct gcagcccccc tacaccaagc ccgtgatcca gaccagacac gacgacgccg
     1261 tgcaccccga gagcggcgcc tatgctagcg tgctgcccgt gtccctgaac gctaccgtgg
     1321 ccgacccttt cctgacccct gccaccctga tggaaggcgt gcggagcacc ggcaaggcca
     1381 tcacccccca gcacatcttc accaacgccg ccctgcggct ggtgttcgac gtgcccaacc
     1441 ccggcctgga cttcgtgatc aacctgacca gcgcccagta ccccctgctg caggacccca
     1501 atggcaccct gtggcgctct gtgctgccta gcaagatcga ggtggtcaag cagagcagca
     1561 agtacaccta cgacgtgggc aacctgaacc tgcggagcta ccagagccag agcgagtccg
     1621 tggaagtggt ggctggcgcc cctgccgccg atgccgagtg tttcaccctg aaggcctgct
     1681 gcgccgtgaa cagcagctac taccagaaat acgagctggt ggacagcccc gtgccccctt
     1741 acgtgtgtac cggcgtgccc atcccttacc tgctgagaca cttcaccttc ggcgtggact
     1801 ccaccgtgaa gttcagagag gccgccgagg gcgccagctg cgagatgaag gccaaccggt
     1861 ggaccgagga aagcgccacc tgtaccgcca atctgaatgc cgtgctggcc ggcgtgaccg
     1921 ccagccctcc tcacagactg cccagcagag ccttcagcct gtgccggcag ggcgacaatc
     1981 tgaccaactc cgagtccggc agcagcccca tccccgtcgt gattaccgcc ttctgcagcg
     2041 agagccggtg catcggctac acccccgcca actacagaag cgacagaagc acctgtctga
     2101 actaccctct gggcttctac acaagcgccg acgagaacaa caccaaccag ttcagcagct
     2161 tcgaggacaa tgccgccgct ttctggccct acctgttcaa cgactgcaag tggatcgacg
     2221 acgtgaccat gtccagcccc accaccatcc aggacatcag cctgcagtgt gatgtgcccg
     2281 cctccaccgg ctggtccctg agctacgaga acttctccgc ccacctgacc gacacaatcg
     2341 cccccctgtt ccccatcggc agacccacaa gagtggccgg ccagtacgcc tcccccaccg
     2401 acaaggacac catcctgttc gagctggaat acgtgtcctt cagctccttc aaggccggca
     2461 gatggacctt ccggatctgc accacccggc agtgcgccac cagcacagtg cctgtggcag
     2521 gcgacaccgg ccactacccc aacgtgatga gagtgaatgt gtccgccagc aagctgggcc
     2581 tgtctgcctc tccacacgag cccgccgtgg gcgtcgaggt gtacagcgcc acaaccccta
     2641 ccacctcccc ctttcggacc ctgctggccc ccagcgtggt gtacgtggac aaggccgcta
     2701 cccccctgag ccaggacaca gtgaacatga ccctggcccg ggaccctccc gagagagtgc
     2761 ctagccctta cgacgccagc accagcggca gaaccaccag ccgggtgctg tgcgagggcg
     2821 actaccggtt cagcaccgtg accaacaact gccagcccct gacagacgag gactgtgcca
     2881 ccaagtaccg gggcaggcgg agcaagttcg accccgtggc cagagcctgc gcctaccctg
     2941 tgcctcccct gaatggcccc ctggtgttta agcccctggc cgagcccaag ccccccagaa
     3001 cctactcccc cgaggaactg cgggccatcc tgaaaacagt gaagctgccc cagttcctgc
     3061 ggaccatgga aaagtcctac gaggattacg agagacagat ggccagatgg gagggcagaa
     3121 gcgtgcggtt ccctccaggc agagccgccg cagccgagga tacagatgtg gtggccgtgg
     3181 ctgccgctga tgagtccgcc gtgacctaca tcaacgagag ccccatgcct ggcgccagag
     3241 gcctgagcgt gacctgtatc gccgccacat gcgtgctgtg gaccgtggca ctgctgcggg
     3301 acgtgctgga aaagctgttc ggcctgggcc cttggcacga ccagcggaga tgcgccttct
     3361 ttgccggcgc tgtgaactgc tggcactggc tgagcggagc ttgcgccgaa ggccacaacg
     3421 aggccagacc tcggcagcgg agggccacct ctaataccac cgccaccccc acaaccgccc
     3481 agcctagcgc ccctagatcc gccacacaga gcggccagtc tggccagcac gccaaccctg
     3541 ccctgcagaa gagaaagggc aagggcgcaa ccttcaccgc cgacacccct ctgcagagca
     3601 ccagaaagag cccccacggc gctccccggg aacctgtgac ctctgcccca tttcagccca
     3661 gggaccccac agccacaccc cagagagaga agcaggacgg cggccagaag cggcatctgc
     3721 ctagaaccca caacagccgg gacgccgagc gcgagcacag acagtctgtg tctgccggca
     3781 ccaccggccc tgccagctct tggggagatg cccggcacta cgtcagcgcc gctcacccac
     3841 ccgcctacga tgtgttcggc ttcagcgagg cccctcacgc cccttttggc tctggcgccg
     3901 gattcgccgc tcccggcgat atgccttttc tgggcggcga ctataccggc cagagagtgt
     3961 ggctgtccga cgagggcgtg ggcctgggat acgtgccagc cgcctttgcc agaccccact
     4021 ccaccagctc ccaccggtgc cacagctgcc ctagctcccc ctaccctacc tccagcccac
     4081 ccagccccga gtggcacccc tttgaggacg tgtaccccac cgtgccctcc ccatgctacg
     4141 ccccctatga tgccccactg cccgagttta gcctgaaccc cagcagcacc tcctaccccg
     4201 gcttcgccag agctgctcct cctagcccct tcgccggctt tgacgccttc ccacctgtgt
     4261 cctccgtgcg gcagtccagc agggtggaag tcctgagcag cgataccgac ggcgcccacg
     4321 gggacaatag agccgctgaa gcccctagcc cagctggcag gaccagacgg cccaacagca
     4381 tggtcggcgc gcccggcagc ctgcaccaca tcctggacgc ccagaagatg ctgtggaacc
     4441 acagggacag gaacctgcct cctctggccc ctctgggccc ccaccaccat caccaccatt
     4501 gagtctagac acgtgattaa ttaaggatcc cccgacctcg acctctggct aataaaggaa
     4561 atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata
     4621 tgggagggca aatcatttgg tcgagatccc tcggagatct ctagctagag gatcgatccc
     4681 cgccccggac gaactaaacc tgactacgac atctctgccc cttcttcgcg gggcagtgca
     4741 tgtaatccct tcagttggtt ggtacaactt gccaactggg ccctgttcca catgtgacac
     4801 ggggggggac caaacacaaa ggggttctct gactgtagtt gacatcctta taaatggatg
     4861 tgcacatttg ccaacactga gtggctttca tcctggagca gactttgcag tctgtggact
     4921 gcaacacaac attgccttta tgtgtaactc ttggctgaag ctcttacacc aatgctgggg
     4981 gacatgtacc tcccaggggc ccaggaagac tacgggaggc tacaccaacg tcaatcagag
     5041 gggcctgtgt agctaccgat aagcggaccc tcaagagggc attagcaata gtgtttataa
     5101 ggcccccttg ttaaccctaa acgggtagca tatgcttccc gggtagtagt atatactatc
     5161 cagactaacc ctaattcaat agcatatgtt acccaacggg aagcatatgc tatcgaatta
     5221 gggttagtaa aagggtccta aggaacagcg atatctccca ccccatgagc tgtcacggtt
     5281 ttatttacat ggggtcagga ttccacgagg gtagtgaacc attttagtca caagggcagt
     5341 ggctgaagat caaggagcgg gcagtgaact ctcctgaatc ttcgcctgct tcttcattct
     5401 ccttcgttta gctaatagaa taactgctga gttgtgaaca gtaaggtgta tgtgaggtgc
     5461 tcgaaaacaa ggtttcaggt gacgccccca gaataaaatt tggacggggg gttcagtggt
     5521 ggcattgtgc tatgacacca atataaccct cacaaacccc ttgggcaata aatactagtg
     5581 taggaatgaa acattctgaa tatctttaac aatagaaatc catggggtgg ggacaagccg
     5641 taaagactgg atgtccatct cacacgaatt tatggctatg ggcaacacat aatcctagtg
     5701 caatatgata ctggggttat taagatgtgt cccaggcagg gaccaagaca ggtgaaccat
     5761 gttgttacac tctatttgta acaaggggaa agagagtgga cgccgacagc agcggactcc
     5821 actggttgtc tctaacaccc ccgaaaatta aacggggctc cacgccaatg gggcccataa
     5881 acaaagacaa gtggccactc ttttttttga aattgtggag tgggggcacg cgtcagcccc
     5941 cacacgccgc cctgcggttt tggactgtaa aataagggtg taataacttg gctgattgta
     6001 accccgctaa ccactgcggt caaaccactt gcccacaaaa ccactaatgg caccccgggg
     6061 aatacctgca taagtaggtg ggcgggccaa gataggggcg cgattgctgc gatctggagg
     6121 acaaattaca cacacttgcg cctgagcgcc aagcacaggg ttgttggtcc tcatattcac
     6181 gaggtcgctg agagcacggt gggctaatgt tgccatgggt agcatatact acccaaatat
     6241 ctggatagca tatgctatcc taatctatat ctgggtagca taggctatcc taatctatat
     6301 ctgggtagca tatgctatcc taatctatat ctgggtagta tatgctatcc taatttatat
     6361 ctgggtagca taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat
     6421 ctgggtagta tatgctatcc taatctgtat ccgggtagca tatgctatcc taatagagat
     6481 tagggtagta tatgctatcc taatttatat ctgggtagca tatactaccc aaatatctgg
     6541 atagcatatg ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg
     6601 gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg
     6661 gtagtatatg ctatcctaat ttatatctgg gtagcatagg ctatcctaat ctatatctgg
     6721 gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat ctgtatccgg
     6781 gtagcatatg ctatcctcat gataagctgt caaacatgag aattttcttg aagacgaaag
     6841 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     6901 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     6961 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     7021 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     7081 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     7141 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     7201 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     7261 gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     7321 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     7381 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     7441 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     7501 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     7561 gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     7621 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     7681 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     7741 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     7801 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     7861 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     7921 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     7981 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     8041 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     8101 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     8161 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     8221 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     8281 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     8341 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     8401 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     8461 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     8521 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     8581 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     8641 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     8701 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     8761 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     8821 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     8881 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     8941 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     9001 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     9061 tacgccaagc tctagctaga ggtcgaccaa ttctcatgtt tgacagctta tcatcgcaga
     9121 tccgggcaac gttgttgcca ttgctgcagg cgcagaactg gtaggtatgg aagatctata
     9181 cattgaatca atattggcaa ttagccatat tagtcattgg ttatatagca taaatcaata
     9241 ttggctattg gccattgcat acgttgtatc tatatcataa tatgtacatt tatattggct
     9301 catgtccaat atgaccgcca tgtt
//
