LOCUS       sequence-274556-       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        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             5262..5279
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    5331..5386
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5385..5404)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(7622..7640)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7708..7812
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7813..7881,7882..8673)
                     /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(8031..8050)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8844..9432
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9333..9352
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9586..9603
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9720..9741
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9756..9761,9762..9779,9780..9786)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9794..9810
                     /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     9799..9821
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9818..9834
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9818..9834
                     /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 gccaccctgg
     1201 gagccgattc tgccaccccc ctgctgtgcg acaacgacgc cacactgtac accaccggcc
     1261 ctgatggcac cgccctgggc atcctgctgt acgagggcgt ggtgccctgc gccagcaacg
     1321 agacaatctt catcacaatc gccaaggacc ccagcccccc caacgccgtg gtggctctga
     1381 gcgagtggtc cttcctggtg tccatcaaca acagagcccc tgccaccttc accctgtacg
     1441 ccatgtgcgg caacaccatc aagtgccaga gcaccctgac ctacgccctg agccagacca
     1501 tggccagcag cagctccagc gacgtgctgc ccctgagcag ccctagctct agcagcgccg
     1561 actctgtgct gcctaacacc acctaccccc tgtgctccca cgcctccaag gccagatcct
     1621 acagcgtggg cgaggtgtac cacgacaact tccccatgga catccaggac ggcagctgcg
     1681 tgtacacaac cagcatcacc ggcgagtcca ccgtgggcac agtggtggtg gccacctccg
     1741 gcctgagcta caccttcagc tcctccaaga cagccgccgt gggctacaac attctgctgg
     1801 gctatatcgt gcggtgcaac ggcatcacag tgtgcgccga caccgtggat gtgctgctgg
     1861 aaagcgccag cgtgaccacc gcccctccaa caagcacacc cgcccctacc cctgcccctc
     1921 ccgagacact gccttggtgc agccaccgga catacgccaa cacctaccgg atcggcgaag
     1981 tgtacttcgg cgacctgctg ctcgacgtga aggacaccga gtgccggtac accgccacca
     2041 tcgaggaccc tagcgtggga acagccgtgg tcgccgagcc tagcgacccc ctgggctacg
     2101 tgtacagcac ctccgacgat accgtgcccc agaacacctc catcccttac accattgcct
     2161 gcaatggcgt gtccatttgc cacggcctgg tggtcatcca cctgacttac gaccctcccg
     2221 cctgccccaa cagcgtcgtg acctatctgc tcgagcctgg cggctccgtg accggcgcca
     2281 tcagcagcgg aacagtgtgt agcagcggcg ccacacctag ccccagcatc ctgagccccg
     2341 tgcccggatt cttcctggac gccagcggcg acttcttctt ctacgccacc gacgacgagg
     2401 ccgaggtcgt ggccatggtg ctgatgaagt gcaacgaaga ggtgctgtgc cagatcaaag
     2461 tgatgtttct ggtggcctac cccaccacag ccccacccac cacctccacc acaaccaccg
     2521 ctgcccccat cgtgatgtgc cccaagctgt tcagatacga ggtgcccacc gggcagagca
     2581 tcagcgatac catcgacccc gtgacaacct ccagctgcaa cttcaccaca tacctgctgg
     2641 tcaactccag cagcgacatc agaggatctc tgctgctgaa cctgctcggc gagttctact
     2701 acaaggcccc caagagccag gccgtggact acgccgctgt ggacgtgtac tgcgccaagg
     2761 ccttcgtgtg ccggacccag ctggtgttcg tggcctatac ccctctgcca accaccagca
     2821 ccaccatgcc tccactgccc ccttgcgcca atgtgtacta ctaccagacc agccctgggg
     2881 tggccatcaa tgcctccctg aacaacatgc ccggccagga catgtgcgcc cacggccggt
     2941 actttaccct gaatacctcc ccacaggccg gcagcgtgaa cgtgtccctg atcggagact
     3001 tcgtgtacct gcctcctaag tatgagggcc agttcagctt catgttcacc atgcactgca
     3061 tgaaccagca gtactgcacc ggcaccgctt atctgctggt gtctaacgag tggaccctga
     3121 ggccagcccc aaccgacagc cctgtgaagc ccgacagcaa cgcctctatc accaacgagc
     3181 agatcacctg tcacggcacc tgtaacgcct ccgcctggaa aacctaccct aaccggaaag
     3241 tgtgggacat cacccctggc gccggatacg ccagaaagga cggccggacc gtgaatgaca
     3301 tgaaggtcac atggcggaac aagagcctgg tcgtgctggt ctacagcctg attggcaacc
     3361 tgggcgtgcg gttccccacc ttcgagccca tcaccacaag ccagcgggcc tacatggaac
     3421 ccaaggactt cgccggaagc gtggccccag gcagccccgg ctttgagatg agctgcctgg
     3481 ccaaccaggg cagaaccggc atgggagagg acgtgtggaa gtggttcagc ctgaccctgg
     3541 acagcggcac aggcggcgtg ggagcctact ataataccgg ccagagctgg tatcagaagt
     3601 tcggcggcaa gcacgtgaac tgcgacatct tcaccgaccc ctgcagatac gccccactgc
     3661 tgacccccgc caactacacc aacaccagcc tgggccggtg gaccgtggac gtcgacgact
     3721 gtgatgccac ctggaccggc gtgttcgagc agagcagcat gggcaagatg gtcaaatacg
     3781 acggctcccc cgtgtggtct ttcgtgggca agctccagct ccagggcacc ctgtatagcg
     3841 aggccgtgca ggccaggtcc tggctggaac ctggccagtt tgacgccctg tacagctccc
     3901 acaacatcct gatcaacctg caccagtttg tggccgtgtc caagaccgga atgcaggacc
     3961 ctctgatcag cgtggacgcc gaattcttca cctacctgga cgacgaaacc caggaccagg
     4021 cctttggcat caacatgctc gtgtatccct tcgtggacag ctacatgacc acctcttacg
     4081 cccgggacag acacgtgaaa ggcttcaagt ggatcctgca ggactggatc agcccaagcg
     4141 acgagcactg ccctacctgt accggcagca aaatgaagtg cgtggcacct ctgcagggcg
     4201 tcgacgtgtc ctacaccgga cctagcttcc ccgagggcga ctgtgccgat ggcgagggca
     4261 gagtgcacct gttcaagggc ccagccacaa tgcccagcga ctgccccgac tccaagatgc
     4321 atgtgttcaa ccagaccggc ttctccccca cccggaactg caataccgcc taccagaaca
     4381 tgaccctgcg gggcatcgtg cctggcagct ctggcctgaa cgagactctg agccgggcct
     4441 tccactacga gggaaccgtg cagctggtcc tgctgatgga cgaccactcc tacaagcggc
     4501 tggacctgca tctgtctatg tacgtgtcca gactgagcaa ggacagcccc cggatgagag
     4561 gcggcctgtc cacctgtaga agcggctcct actggcccgt gctggatcct ctgggcagct
     4621 ccctggcctc cagccccttt cctctgtccg tggctgccgt gacacccctg tgtatcgacg
     4681 acctgtcctc tagctacggc cccaacgact gggccctgtt cgccgtgaac atgcctggcg
     4741 tgaagccaag cgaagtgacc gtgcggagcg tgtacgtgca gcacaacaac tcccggatct
     4801 acctgatcta caaggatccc agaagcggag ccgcctccat tcccaactcc gacgccgacg
     4861 gcgattggtg gcagtacaag tacccctttc tggccttccg ggacctggaa gcccacgtga
     4921 agaacaattc catcacctac agcacactga tggaaagctc cgccgctgcc gccgacgtgg
     4981 tgtacgcctt caccttcatc ccaggcgctc tgggagtgga taccgacatc gaggtggtgg
     5041 tcgaagccct gatccagccc agcgccgcag ccgagagcac cacagagttc cggcgggtgc
     5101 tgcacatcga tccactgctc acacagctga gcagacaggg ccctccacct ccctatagca
     5161 gctctggcgc gcccggcagc ctgcaccaca tcctggacgc ccagaagatg ctgtggaacc
     5221 acagggacag gaacctgcct cctctggccc ctctgggccc ccaccaccat caccaccatt
     5281 gagtctagac acgtgattaa ttaaggatcc cccgacctcg acctctggct aataaaggaa
     5341 atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata
     5401 tgggagggca aatcatttgg tcgagatccc tcggagatct ctagctagag gatcgatccc
     5461 cgccccggac gaactaaacc tgactacgac atctctgccc cttcttcgcg gggcagtgca
     5521 tgtaatccct tcagttggtt ggtacaactt gccaactggg ccctgttcca catgtgacac
     5581 ggggggggac caaacacaaa ggggttctct gactgtagtt gacatcctta taaatggatg
     5641 tgcacatttg ccaacactga gtggctttca tcctggagca gactttgcag tctgtggact
     5701 gcaacacaac attgccttta tgtgtaactc ttggctgaag ctcttacacc aatgctgggg
     5761 gacatgtacc tcccaggggc ccaggaagac tacgggaggc tacaccaacg tcaatcagag
     5821 gggcctgtgt agctaccgat aagcggaccc tcaagagggc attagcaata gtgtttataa
     5881 ggcccccttg ttaaccctaa acgggtagca tatgcttccc gggtagtagt atatactatc
     5941 cagactaacc ctaattcaat agcatatgtt acccaacggg aagcatatgc tatcgaatta
     6001 gggttagtaa aagggtccta aggaacagcg atatctccca ccccatgagc tgtcacggtt
     6061 ttatttacat ggggtcagga ttccacgagg gtagtgaacc attttagtca caagggcagt
     6121 ggctgaagat caaggagcgg gcagtgaact ctcctgaatc ttcgcctgct tcttcattct
     6181 ccttcgttta gctaatagaa taactgctga gttgtgaaca gtaaggtgta tgtgaggtgc
     6241 tcgaaaacaa ggtttcaggt gacgccccca gaataaaatt tggacggggg gttcagtggt
     6301 ggcattgtgc tatgacacca atataaccct cacaaacccc ttgggcaata aatactagtg
     6361 taggaatgaa acattctgaa tatctttaac aatagaaatc catggggtgg ggacaagccg
     6421 taaagactgg atgtccatct cacacgaatt tatggctatg ggcaacacat aatcctagtg
     6481 caatatgata ctggggttat taagatgtgt cccaggcagg gaccaagaca ggtgaaccat
     6541 gttgttacac tctatttgta acaaggggaa agagagtgga cgccgacagc agcggactcc
     6601 actggttgtc tctaacaccc ccgaaaatta aacggggctc cacgccaatg gggcccataa
     6661 acaaagacaa gtggccactc ttttttttga aattgtggag tgggggcacg cgtcagcccc
     6721 cacacgccgc cctgcggttt tggactgtaa aataagggtg taataacttg gctgattgta
     6781 accccgctaa ccactgcggt caaaccactt gcccacaaaa ccactaatgg caccccgggg
     6841 aatacctgca taagtaggtg ggcgggccaa gataggggcg cgattgctgc gatctggagg
     6901 acaaattaca cacacttgcg cctgagcgcc aagcacaggg ttgttggtcc tcatattcac
     6961 gaggtcgctg agagcacggt gggctaatgt tgccatgggt agcatatact acccaaatat
     7021 ctggatagca tatgctatcc taatctatat ctgggtagca taggctatcc taatctatat
     7081 ctgggtagca tatgctatcc taatctatat ctgggtagta tatgctatcc taatttatat
     7141 ctgggtagca taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat
     7201 ctgggtagta tatgctatcc taatctgtat ccgggtagca tatgctatcc taatagagat
     7261 tagggtagta tatgctatcc taatttatat ctgggtagca tatactaccc aaatatctgg
     7321 atagcatatg ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg
     7381 gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg
     7441 gtagtatatg ctatcctaat ttatatctgg gtagcatagg ctatcctaat ctatatctgg
     7501 gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat ctgtatccgg
     7561 gtagcatatg ctatcctcat gataagctgt caaacatgag aattttcttg aagacgaaag
     7621 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     7681 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     7741 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     7801 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     7861 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     7921 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     7981 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     8041 gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     8101 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     8161 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     8221 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     8281 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     8341 gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     8401 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     8461 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     8521 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     8581 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     8641 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     8701 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     8761 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     8821 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     8881 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     8941 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     9001 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     9061 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     9121 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     9181 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     9241 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     9301 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     9361 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     9421 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     9481 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     9541 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     9601 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     9661 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     9721 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     9781 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     9841 tacgccaagc tctagctaga ggtcgaccaa ttctcatgtt tgacagctta tcatcgcaga
     9901 tccgggcaac gttgttgcca ttgctgcagg cgcagaactg gtaggtatgg aagatctata
     9961 cattgaatca atattggcaa ttagccatat tagtcattgg ttatatagca taaatcaata
    10021 ttggctattg gccattgcat acgttgtatc tatatcataa tatgtacatt tatattggct
    10081 catgtccaat atgaccgcca tgtt
//
