LOCUS       sequence-274892-        8424 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 8424)
  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 8424)
  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: D45_Ld_343960
FEATURES             Location/Qualifiers
     source          1..8424
                     /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             3582..3599
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3651..3706
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3705..3724)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5942..5960)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6028..6132
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6133..6201,6202..6993)
                     /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(6351..6370)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7164..7752
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7653..7672
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7906..7923
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8040..8061
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8076..8081,8082..8099,8100..8106)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8114..8130
                     /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     8119..8141
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8138..8154
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8138..8154
                     /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 gccaccatcc
     1201 acgaggatga gcagggcctg cgggactgga tcatgcacct cgtgggcaac gtgcaggact
     1261 ctgccgccca ggtggccacc aacccccagc tgctgtacgt ggcctctgag gatggcgccg
     1321 tggccgccat tgccgtggga gcctatggcg gcctgaacct gacctggcgg cagatcagca
     1381 gcgccacccc tctgtgtgtg gccgctggac ctcaggccgt gctgaccgtg aacaacgccg
     1441 gcgtggccac actgatgagc cccaccaccg gcagcatcga agtgacctac gccctgcaga
     1501 ccgtgaccac cacactgcag gccgctgcct gtagcgtggt gtccggaaag gccgtggtgg
     1561 tcacctacga cggggccacc ctgaagagat tcgagttcag caccgccagc gaggaacaga
     1621 gcatccccgt ggctgcctac gccagcgcct ctggcgaggt gtccaagatg tacgtcgccg
     1681 gcagcgtgct gctggtcaac cacggcagcg acagcgccga cgtgctgaga ctggacaacc
     1741 tggccaaaga acgcagcgtg gccggaatgg ccacctctat cgccgccgac ggccacttca
     1801 ccacccggga tgccgccaca gtgcggagct tccctaccaa cggcgctgcc agcgaagtgg
     1861 cctgcagcga ctgtggaatc gccgtcgtgc ggaacgccca caccggcgaa cctaccggcg
     1921 tcgtcagagt ggaaacacag gtggacatgc tgcggatcac ctaccccaac agcgtcgtga
     1981 ccatccccaa catctgcagc agcatcaccg cctaccccct gctggccttc gagaaggacg
     2041 acggcgacgt gatcgtgctg atcaagacag gcgcccacaa cctgctgctc gtgtccgccg
     2101 ctgaaggcct cgtgtggcgg agatttgagg ccctggctaa cgtggcagcc gccgctatcg
     2161 tggaacccat ggaagccctg gaccacttcc acttcaacaa gaacatcctg ctggtgtcca
     2221 gattcggcac cctgtacagc atccctattg ccggcatggg agcccaggtc gactacatca
     2281 aggacttcag ccaggaactg gctgccgcca tgaaggcccc cagcatggct aaggtgcagg
     2341 tccgagagct ggccgtgcgg gatgatggca gaaccgctgt ggtgcacgcc gagtctggcg
     2401 ccacaagcgg ccacatcctg atcgacctgg ccgagcggat ggtcaccagc gtgacaacat
     2461 gcgagaacgc tgtgctgagc acccccgagc tggaagtggc tgctgacctg agcgtgcggg
     2521 gcagcctgag ccacggcgtg ttctacacct atgccagcca cgccgaaagc ggcaccatcg
     2581 agggctactc cgtgtctgct gccgcaggcg ccagacccac ctgggtggtg cagatgccta
     2641 gcgccattgt ggcccacgcc gctggcagcg agcctcggag aaccgacgtg gtcaacaacc
     2701 tgagagtgta ccccaatatc agcggcaggg aaatggtgga agaagtgcgg cggacatacc
     2761 ccatgcggaa tgtgatcgct gtggctcact acgagccctc cgaggaagaa ctgcccagcc
     2821 tggtcatcac agccatcgat gtggtcacag gctccatcct ggccacaacc agacacgcca
     2881 acgtcgaggg ggacgtgaag atggtcattg tggaacacgc catcgtgtac tactacctgg
     2941 acgccaagaa gatgcggtac tgcttcggcg tgtgggagct gttcgaggac gagaacagcc
     3001 ctgtggtgtc taagacagcc ggcgctacca tccctcagat tatcgccagc ttcttcgtga
     3061 acaccaagag agagttctcc agccgggcca ccaggccccc catcgtggtc gtgtctaccc
     3121 tgggcagctt tggcggccct ctggccgata tgggagtgac cacaagctac aacgctatcg
     3181 cccggaagtc cctggtgctg gcttacgcca caggccgggt cgtcgtggtg gaactgagac
     3241 acctcctggc cgggaaccag atgcccctgc ctggggccaa ggaccggcag atgagccatc
     3301 tgctgctgcc tagcttcctg ttcgcctccc acaagtaccg ggtggccttc cccagaaaga
     3361 tcaccaccga gcccaccctg ctcgagagca gctgtcatgt ggtcgtcagc ggcctggatc
     3421 tgttctacgt gcggtccagc agcggcaagc ccttcgacct gctgaacagc gactttaaca
     3481 agcctggcgc gcccggcagc ctgcaccaca tcctggacgc ccagaagatg ctgtggaacc
     3541 acagggacag gaacctgcct cctctggccc ctctgggccc ccaccaccat caccaccatt
     3601 gagtctagac acgtgattaa ttaaggatcc cccgacctcg acctctggct aataaaggaa
     3661 atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata
     3721 tgggagggca aatcatttgg tcgagatccc tcggagatct ctagctagag gatcgatccc
     3781 cgccccggac gaactaaacc tgactacgac atctctgccc cttcttcgcg gggcagtgca
     3841 tgtaatccct tcagttggtt ggtacaactt gccaactggg ccctgttcca catgtgacac
     3901 ggggggggac caaacacaaa ggggttctct gactgtagtt gacatcctta taaatggatg
     3961 tgcacatttg ccaacactga gtggctttca tcctggagca gactttgcag tctgtggact
     4021 gcaacacaac attgccttta tgtgtaactc ttggctgaag ctcttacacc aatgctgggg
     4081 gacatgtacc tcccaggggc ccaggaagac tacgggaggc tacaccaacg tcaatcagag
     4141 gggcctgtgt agctaccgat aagcggaccc tcaagagggc attagcaata gtgtttataa
     4201 ggcccccttg ttaaccctaa acgggtagca tatgcttccc gggtagtagt atatactatc
     4261 cagactaacc ctaattcaat agcatatgtt acccaacggg aagcatatgc tatcgaatta
     4321 gggttagtaa aagggtccta aggaacagcg atatctccca ccccatgagc tgtcacggtt
     4381 ttatttacat ggggtcagga ttccacgagg gtagtgaacc attttagtca caagggcagt
     4441 ggctgaagat caaggagcgg gcagtgaact ctcctgaatc ttcgcctgct tcttcattct
     4501 ccttcgttta gctaatagaa taactgctga gttgtgaaca gtaaggtgta tgtgaggtgc
     4561 tcgaaaacaa ggtttcaggt gacgccccca gaataaaatt tggacggggg gttcagtggt
     4621 ggcattgtgc tatgacacca atataaccct cacaaacccc ttgggcaata aatactagtg
     4681 taggaatgaa acattctgaa tatctttaac aatagaaatc catggggtgg ggacaagccg
     4741 taaagactgg atgtccatct cacacgaatt tatggctatg ggcaacacat aatcctagtg
     4801 caatatgata ctggggttat taagatgtgt cccaggcagg gaccaagaca ggtgaaccat
     4861 gttgttacac tctatttgta acaaggggaa agagagtgga cgccgacagc agcggactcc
     4921 actggttgtc tctaacaccc ccgaaaatta aacggggctc cacgccaatg gggcccataa
     4981 acaaagacaa gtggccactc ttttttttga aattgtggag tgggggcacg cgtcagcccc
     5041 cacacgccgc cctgcggttt tggactgtaa aataagggtg taataacttg gctgattgta
     5101 accccgctaa ccactgcggt caaaccactt gcccacaaaa ccactaatgg caccccgggg
     5161 aatacctgca taagtaggtg ggcgggccaa gataggggcg cgattgctgc gatctggagg
     5221 acaaattaca cacacttgcg cctgagcgcc aagcacaggg ttgttggtcc tcatattcac
     5281 gaggtcgctg agagcacggt gggctaatgt tgccatgggt agcatatact acccaaatat
     5341 ctggatagca tatgctatcc taatctatat ctgggtagca taggctatcc taatctatat
     5401 ctgggtagca tatgctatcc taatctatat ctgggtagta tatgctatcc taatttatat
     5461 ctgggtagca taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat
     5521 ctgggtagta tatgctatcc taatctgtat ccgggtagca tatgctatcc taatagagat
     5581 tagggtagta tatgctatcc taatttatat ctgggtagca tatactaccc aaatatctgg
     5641 atagcatatg ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg
     5701 gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg
     5761 gtagtatatg ctatcctaat ttatatctgg gtagcatagg ctatcctaat ctatatctgg
     5821 gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat ctgtatccgg
     5881 gtagcatatg ctatcctcat gataagctgt caaacatgag aattttcttg aagacgaaag
     5941 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     6001 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     6061 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     6121 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     6181 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     6241 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     6301 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     6361 gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     6421 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     6481 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     6541 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     6601 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     6661 gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     6721 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     6781 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     6841 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     6901 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     6961 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     7021 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     7081 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     7141 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     7201 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     7261 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     7321 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     7381 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     7441 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     7501 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     7561 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     7621 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     7681 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     7741 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     7801 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     7861 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     7921 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     7981 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     8041 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     8101 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     8161 tacgccaagc tctagctaga ggtcgaccaa ttctcatgtt tgacagctta tcatcgcaga
     8221 tccgggcaac gttgttgcca ttgctgcagg cgcagaactg gtaggtatgg aagatctata
     8281 cattgaatca atattggcaa ttagccatat tagtcattgg ttatatagca taaatcaata
     8341 ttggctattg gccattgcat acgttgtatc tatatcataa tatgtacatt tatattggct
     8401 catgtccaat atgaccgcca tgtt
//
