LOCUS       sequence-274670-        6324 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 6324)
  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 6324)
  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: D91_Ld_352550
FEATURES             Location/Qualifiers
     source          1..6324
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     201..225
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     706..725
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      748..757
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1102..1119
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1171..1226
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1225..1244)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3462..3480)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3548..3652
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3653..3721,3722..4513)
                     /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(3871..3890)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4684..5272
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5173..5192
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5426..5443
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5560..5581
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5596..5601,5602..5619,5620..5626)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5634..5650
                     /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     5639..5661
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5658..5674
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5658..5674
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        5945..6324
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta caccaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcctcac tctcttccgc
      241 atcgctgtct gcgagggcca gctgttgggc tcgcggttga ggacaaactc ttcgcggtct
      301 ttccagtact cttggatcgg aaacccgtcg gcctccgaac ggtactccgc caccgaggga
      361 cctgagcgag tccgcatcga ccggatcgga aaacctctcg agaaaggcgt ctaaccagtc
      421 acagtcgcaa ggtaggctga gcaccgtggc gggcggcagc gggtggcggt cggggttgtt
      481 tctggcggag gtgctgctga tgatgtaatt aaagtaggcg gtcttgagac ggcggatggt
      541 cgaggtgagg tgtggcaggc ttgagatcca gctgttgggg tgagtactcc ctctcaaaag
      601 cgggcattac ttctgcgcta agattgtcag tttccaaaaa cgaggaggat ttgatattca
      661 cctggcccga tctggccata cacttgagtg acaatgacat ccactttgcc tttctctcca
      721 caggtgtcca ctcccaggtc caagtttgcc accatggagt ttcagaccca ggtactcatg
      781 tccctgctgc tctgcatgtc tggtgcggcc gccagatttc tgtctaagtt tggactggcc
      841 acacctcagg ctctgcaagg cggaattgct cctcagcagg acaccgcctt caacgacatg
      901 gtgcctagca gcttcgacgc cgacttcagc tctaatggaa acggcggcgt cagatccgtg
      961 tccagcatga agctgcagat cgccagcctg ctgcacagca tcagaggcgc gcccggcagc
     1021 ctgcaccaca tcctggacgc ccagaagatg ctgtggaacc acagggacag gaacctgcct
     1081 cctctggccc ctctgggccc ccaccaccat caccaccatt gagtctagac acgtgattaa
     1141 ttaaggatcc cccgacctcg acctctggct aataaaggaa atttattttc attgcaatag
     1201 tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttgg
     1261 tcgagatccc tcggagatct ctagctagag gatcgatccc cgccccggac gaactaaacc
     1321 tgactacgac atctctgccc cttcttcgcg gggcagtgca tgtaatccct tcagttggtt
     1381 ggtacaactt gccaactggg ccctgttcca catgtgacac ggggggggac caaacacaaa
     1441 ggggttctct gactgtagtt gacatcctta taaatggatg tgcacatttg ccaacactga
     1501 gtggctttca tcctggagca gactttgcag tctgtggact gcaacacaac attgccttta
     1561 tgtgtaactc ttggctgaag ctcttacacc aatgctgggg gacatgtacc tcccaggggc
     1621 ccaggaagac tacgggaggc tacaccaacg tcaatcagag gggcctgtgt agctaccgat
     1681 aagcggaccc tcaagagggc attagcaata gtgtttataa ggcccccttg ttaaccctaa
     1741 acgggtagca tatgcttccc gggtagtagt atatactatc cagactaacc ctaattcaat
     1801 agcatatgtt acccaacggg aagcatatgc tatcgaatta gggttagtaa aagggtccta
     1861 aggaacagcg atatctccca ccccatgagc tgtcacggtt ttatttacat ggggtcagga
     1921 ttccacgagg gtagtgaacc attttagtca caagggcagt ggctgaagat caaggagcgg
     1981 gcagtgaact ctcctgaatc ttcgcctgct tcttcattct ccttcgttta gctaatagaa
     2041 taactgctga gttgtgaaca gtaaggtgta tgtgaggtgc tcgaaaacaa ggtttcaggt
     2101 gacgccccca gaataaaatt tggacggggg gttcagtggt ggcattgtgc tatgacacca
     2161 atataaccct cacaaacccc ttgggcaata aatactagtg taggaatgaa acattctgaa
     2221 tatctttaac aatagaaatc catggggtgg ggacaagccg taaagactgg atgtccatct
     2281 cacacgaatt tatggctatg ggcaacacat aatcctagtg caatatgata ctggggttat
     2341 taagatgtgt cccaggcagg gaccaagaca ggtgaaccat gttgttacac tctatttgta
     2401 acaaggggaa agagagtgga cgccgacagc agcggactcc actggttgtc tctaacaccc
     2461 ccgaaaatta aacggggctc cacgccaatg gggcccataa acaaagacaa gtggccactc
     2521 ttttttttga aattgtggag tgggggcacg cgtcagcccc cacacgccgc cctgcggttt
     2581 tggactgtaa aataagggtg taataacttg gctgattgta accccgctaa ccactgcggt
     2641 caaaccactt gcccacaaaa ccactaatgg caccccgggg aatacctgca taagtaggtg
     2701 ggcgggccaa gataggggcg cgattgctgc gatctggagg acaaattaca cacacttgcg
     2761 cctgagcgcc aagcacaggg ttgttggtcc tcatattcac gaggtcgctg agagcacggt
     2821 gggctaatgt tgccatgggt agcatatact acccaaatat ctggatagca tatgctatcc
     2881 taatctatat ctgggtagca taggctatcc taatctatat ctgggtagca tatgctatcc
     2941 taatctatat ctgggtagta tatgctatcc taatttatat ctgggtagca taggctatcc
     3001 taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagta tatgctatcc
     3061 taatctgtat ccgggtagca tatgctatcc taatagagat tagggtagta tatgctatcc
     3121 taatttatat ctgggtagca tatactaccc aaatatctgg atagcatatg ctatcctaat
     3181 ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagcatagg ctatcctaat
     3241 ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat
     3301 ttatatctgg gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat
     3361 ctatatctgg gtagtatatg ctatcctaat ctgtatccgg gtagcatatg ctatcctcat
     3421 gataagctgt caaacatgag aattttcttg aagacgaaag ggcctcgtga tacgcctatt
     3481 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     3541 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3601 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     3661 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     3721 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     3781 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     3841 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga
     3901 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     3961 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     4021 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     4081 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     4141 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc
     4201 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     4261 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     4321 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     4381 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     4441 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     4501 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     4561 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     4621 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     4681 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     4741 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     4801 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     4861 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4921 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4981 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     5041 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     5101 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     5161 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     5221 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     5281 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     5341 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     5401 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     5461 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     5521 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     5581 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     5641 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc tctagctaga
     5701 ggtcgaccaa ttctcatgtt tgacagctta tcatcgcaga tccgggcaac gttgttgcca
     5761 ttgctgcagg cgcagaactg gtaggtatgg aagatctata cattgaatca atattggcaa
     5821 ttagccatat tagtcattgg ttatatagca taaatcaata ttggctattg gccattgcat
     5881 acgttgtatc tatatcataa tatgtacatt tatattggct catgtccaat atgaccgcca
     5941 tgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
     6001 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
     6061 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
     6121 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     6181 catcaagtgt atcatatgcc aagtccgccc cctattgacg tcaatgacgg taaatggccc
     6241 gcctggcatt atgcccagta catgacctta cgggactttc ctacttggca gtacatctac
     6301 gtattagtca tcgctattac catg
//
