LOCUS       sequence-274877-        6759 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 6759)
  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 6759)
  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: D66_Ld_080470
FEATURES             Location/Qualifiers
     source          1..6759
                     /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             1537..1554
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1606..1661
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1660..1679)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3897..3915)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3983..4087
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4088..4156,4157..4948)
                     /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(4306..4325)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5119..5707
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5608..5627
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5861..5878
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5995..6016
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6031..6036,6037..6054,6055..6061)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6069..6085
                     /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     6074..6096
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6093..6109
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6093..6109
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        6380..6759
                     /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 gccggtgaca atgtgatact ttccctgcca
      841 agtggtgtgc gccttgcgcg cgcggacgga cctatcacta aagagcagtg tttcattttg
      901 aatcctaagc ctaccgtaac tagttgtaac gcctacctgc caccaatcgt ggatctgtat
      961 ggcatgctcc cctacgataa gacacctgct tatacaaatt gcgtcaagga gcggtgcctc
     1021 tgtacaggcg ctgccacgtc aacagctatc aagacctcag gattcttttg taatagctct
     1081 ggatggatgg agagcgggta cacgacctgt aatgttctgg tagattgctt ctctaagttc
     1141 tggagatgta tcaacaaggc cctgtataca cggtacctcg aggatcgggg taacctggac
     1201 cccacagaag tggacctgat ggcagatatg atttctcacg ggaatcgccc tggagaaacc
     1261 ttcgacatta cagatatctt ccggtcttgt cgccttatga tgtgtgccgc tgcgaactcc
     1321 cggcagaatt gtggactggt cacctgtctg ccaaattaca cccagtgtga tgaatacatc
     1381 cgtccgcccc cgctgcccta tacacatcag ctgtgcactc agggctgtcg ggccgccctg
     1441 ggcgcgcccg gcagcctgca ccacatcctg gacgcccaga agatgctgtg gaaccacagg
     1501 gacaggaacc tgcctcctct ggcccctctg ggcccccacc accatcacca ccattgagtc
     1561 tagacacgtg attaattaag gatcccccga cctcgacctc tggctaataa aggaaattta
     1621 ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga
     1681 gggcaaatca tttggtcgag atccctcgga gatctctagc tagaggatcg atccccgccc
     1741 cggacgaact aaacctgact acgacatctc tgccccttct tcgcggggca gtgcatgtaa
     1801 tcccttcagt tggttggtac aacttgccaa ctgggccctg ttccacatgt gacacggggg
     1861 gggaccaaac acaaaggggt tctctgactg tagttgacat ccttataaat ggatgtgcac
     1921 atttgccaac actgagtggc tttcatcctg gagcagactt tgcagtctgt ggactgcaac
     1981 acaacattgc ctttatgtgt aactcttggc tgaagctctt acaccaatgc tgggggacat
     2041 gtacctccca ggggcccagg aagactacgg gaggctacac caacgtcaat cagaggggcc
     2101 tgtgtagcta ccgataagcg gaccctcaag agggcattag caatagtgtt tataaggccc
     2161 ccttgttaac cctaaacggg tagcatatgc ttcccgggta gtagtatata ctatccagac
     2221 taaccctaat tcaatagcat atgttaccca acgggaagca tatgctatcg aattagggtt
     2281 agtaaaaggg tcctaaggaa cagcgatatc tcccacccca tgagctgtca cggttttatt
     2341 tacatggggt caggattcca cgagggtagt gaaccatttt agtcacaagg gcagtggctg
     2401 aagatcaagg agcgggcagt gaactctcct gaatcttcgc ctgcttcttc attctccttc
     2461 gtttagctaa tagaataact gctgagttgt gaacagtaag gtgtatgtga ggtgctcgaa
     2521 aacaaggttt caggtgacgc ccccagaata aaatttggac ggggggttca gtggtggcat
     2581 tgtgctatga caccaatata accctcacaa accccttggg caataaatac tagtgtagga
     2641 atgaaacatt ctgaatatct ttaacaatag aaatccatgg ggtggggaca agccgtaaag
     2701 actggatgtc catctcacac gaatttatgg ctatgggcaa cacataatcc tagtgcaata
     2761 tgatactggg gttattaaga tgtgtcccag gcagggacca agacaggtga accatgttgt
     2821 tacactctat ttgtaacaag gggaaagaga gtggacgccg acagcagcgg actccactgg
     2881 ttgtctctaa cacccccgaa aattaaacgg ggctccacgc caatggggcc cataaacaaa
     2941 gacaagtggc cactcttttt tttgaaattg tggagtgggg gcacgcgtca gcccccacac
     3001 gccgccctgc ggttttggac tgtaaaataa gggtgtaata acttggctga ttgtaacccc
     3061 gctaaccact gcggtcaaac cacttgccca caaaaccact aatggcaccc cggggaatac
     3121 ctgcataagt aggtgggcgg gccaagatag gggcgcgatt gctgcgatct ggaggacaaa
     3181 ttacacacac ttgcgcctga gcgccaagca cagggttgtt ggtcctcata ttcacgaggt
     3241 cgctgagagc acggtgggct aatgttgcca tgggtagcat atactaccca aatatctgga
     3301 tagcatatgc tatcctaatc tatatctggg tagcataggc tatcctaatc tatatctggg
     3361 tagcatatgc tatcctaatc tatatctggg tagtatatgc tatcctaatt tatatctggg
     3421 tagcataggc tatcctaatc tatatctggg tagcatatgc tatcctaatc tatatctggg
     3481 tagtatatgc tatcctaatc tgtatccggg tagcatatgc tatcctaata gagattaggg
     3541 tagtatatgc tatcctaatt tatatctggg tagcatatac tacccaaata tctggatagc
     3601 atatgctatc ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagc
     3661 ataggctatc ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt
     3721 atatgctatc ctaatttata tctgggtagc ataggctatc ctaatctata tctgggtagc
     3781 atatgctatc ctaatctata tctgggtagt atatgctatc ctaatctgta tccgggtagc
     3841 atatgctatc ctcatgataa gctgtcaaac atgagaattt tcttgaagac gaaagggcct
     3901 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     3961 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     4021 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     4081 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     4141 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     4201 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     4261 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     4321 attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     4381 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     4441 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     4501 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     4561 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     4621 cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     4681 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     4741 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     4801 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     4861 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     4921 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     4981 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     5041 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     5101 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     5161 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5221 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     5281 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     5341 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     5401 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     5461 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     5521 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     5581 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     5641 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     5701 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     5761 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     5821 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     5881 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     5941 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     6001 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     6061 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
     6121 caagctctag ctagaggtcg accaattctc atgtttgaca gcttatcatc gcagatccgg
     6181 gcaacgttgt tgccattgct gcaggcgcag aactggtagg tatggaagat ctatacattg
     6241 aatcaatatt ggcaattagc catattagtc attggttata tagcataaat caatattggc
     6301 tattggccat tgcatacgtt gtatctatat cataatatgt acatttatat tggctcatgt
     6361 ccaatatgac cgccatgttg acattgatta ttgactagtt attaatagta atcaattacg
     6421 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
     6481 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     6541 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     6601 gcccacttgg cagtacatca agtgtatcat atgccaagtc cgccccctat tgacgtcaat
     6661 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttacggga ctttcctact
     6721 tggcagtaca tctacgtatt agtcatcgct attaccatg
//
