LOCUS       sequence-262353-        7602 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 7602)
  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 7602)
  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: D61_Ld_351340
FEATURES             Location/Qualifiers
     source          1..7602
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        7..386
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        387..590
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     541..561
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     587..611
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     1092..1111
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1134..1143
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2766..2783
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2835..2890
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2889..2908)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5126..5144)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5212..5316
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5317..5385,5386..6177)
                     /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(5535..5554)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6348..6936
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6837..6856
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7090..7107
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7224..7245
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7260..7265,7266..7283,7284..7290)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7298..7314
                     /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     7303..7325
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7322..7338
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7322..7338
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 catgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
       61 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      121 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      181 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      241 tacatcaagt gtatcatatg ccaagtccgc cccctattga cgtcaatgac ggtaaatggc
      301 ccgcctggca ttatgcccag tacatgacct tacgggactt tcctacttgg cagtacatct
      361 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacacc aatgggcgtg
      421 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      481 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taataacccc gccccgttga
      541 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga
      601 accgtcagat cctcactctc ttccgcatcg ctgtctgcga gggccagctg ttgggctcgc
      661 ggttgaggac aaactcttcg cggtctttcc agtactcttg gatcggaaac ccgtcggcct
      721 ccgaacggta ctccgccacc gagggacctg agcgagtccg catcgaccgg atcggaaaac
      781 ctctcgagaa aggcgtctaa ccagtcacag tcgcaaggta ggctgagcac cgtggcgggc
      841 ggcagcgggt ggcggtcggg gttgtttctg gcggaggtgc tgctgatgat gtaattaaag
      901 taggcggtct tgagacggcg gatggtcgag gtgaggtgtg gcaggcttga gatccagctg
      961 ttggggtgag tactccctct caaaagcggg cattacttct gcgctaagat tgtcagtttc
     1021 caaaaacgag gaggatttga tattcacctg gcccgatctg gccatacact tgagtgacaa
     1081 tgacatccac tttgcctttc tctccacagg tgtccactcc caggtccaag tttgccacca
     1141 tggagtttca gacccaggta ctcatgtccc tgctgctctg catgtctggt gcggccgccg
     1201 tggaagatgt gggagaggat gccagcgccg tgctggaact gatcagcgag ggcagcacca
     1261 ccacctacaa gatccccgac agcatggtgg tgctgaacaa cgccaacttc gagagctacc
     1321 tgttccccag caagcgggcc acccccagag cctttctggt gctgtgctac agcccctggt
     1381 gcccccactg caagagcctg ctgccccagt tcctgaacgc cagcatgcag ctggacctga
     1441 tgaaggtgcc ccacagcaac ttcgccgtgg tggacgtgca gaaaaacacc gccgtgtccg
     1501 agtacttcga cgtggaacgg ttccccaccc tgctgtacac caccggcaag ggccggcagt
     1561 ggcacctgta cgagggcggc aatacccagc agggcttcat gcagttcagc acctacctgc
     1621 agaacgccat ggacaccggc agcttcagcg aggacgtgac cgacgtgtcc cacttcaacg
     1681 aggtggaaga gaagtccggc accacccggg tgccctgcta cgtgtacgtg cccgccacaa
     1741 gcagcagcgc ccctgagagc cagagaaccg cccactggtc ccacgccatt gacggcgctg
     1801 ccagcgtgtc caacatcaga ttcgccgtga tctacgagaa gtcccaggcc gagggatggg
     1861 ccgagcacgc cagcgacaag tacaagaaag tggtggaaaa ggccaaggcc tgcgtggccg
     1921 ctggcaaggc ttctggacct ggcggagagg ccctggtggt gttcagcgac cggtacagag
     1981 agccccactg ctacagcggc ccctgggtgg aagaaagaag cgtggccaga agcagcaagc
     2041 accggaccag acaggtggac gccgacaccc tgaccatgtc taccagcctg gaaaacttcc
     2101 tggccctgca cggcttccac gccgtcgagg atgcctccag cgccatgttc gccaccctgg
     2161 cctactaccc caagaactac ctgggcgtcg tgatgaccaa ccggcccatc gacgacaagg
     2221 acatggactt cgtgcccgtg ctgagagaga tcacacaggc cgagaacgcc gccctggaaa
     2281 agaagcacgg cagcgacctg cccatcgagg aagagatgcg gacccccaga gtgtcctggt
     2341 cctacatcga cgtggtggaa tacgaagtgt ggcggagcag atacgacatc gagctggacc
     2401 agctgcccgc cgtgatgatc atcgacacca agcgggaccg gttcttcaag atgagaaccc
     2461 acgtgcccag attcgaggcc atcaagatgg acaccccctg gaaagtgggc ggcgagcagc
     2521 agcagctgat cgcccagttc gcccaggacg tgctggccga cgcctacaag gcccagaaac
     2581 tgtccgtggc tggcgccgtg gccgaatacc tgagccacta ccccggcttc gccctgatgt
     2641 acgaggccct gaactacgag gacttcgtgg gcgcgcccgg cagcctgcac cacatcctgg
     2701 acgcccagaa gatgctgtgg aaccacaggg acaggaacct gcctcctctg gcccctctgg
     2761 gcccccacca ccatcaccac cattgagtct agacacgtga ttaattaagg atcccccgac
     2821 ctcgacctct ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt
     2881 gtgtctctca ctcggaagga catatgggag ggcaaatcat ttggtcgaga tccctcggag
     2941 atctctagct agaggatcga tccccgcccc ggacgaacta aacctgacta cgacatctct
     3001 gccccttctt cgcggggcag tgcatgtaat cccttcagtt ggttggtaca acttgccaac
     3061 tgggccctgt tccacatgtg acacgggggg ggaccaaaca caaaggggtt ctctgactgt
     3121 agttgacatc cttataaatg gatgtgcaca tttgccaaca ctgagtggct ttcatcctgg
     3181 agcagacttt gcagtctgtg gactgcaaca caacattgcc tttatgtgta actcttggct
     3241 gaagctctta caccaatgct gggggacatg tacctcccag gggcccagga agactacggg
     3301 aggctacacc aacgtcaatc agaggggcct gtgtagctac cgataagcgg accctcaaga
     3361 gggcattagc aatagtgttt ataaggcccc cttgttaacc ctaaacgggt agcatatgct
     3421 tcccgggtag tagtatatac tatccagact aaccctaatt caatagcata tgttacccaa
     3481 cgggaagcat atgctatcga attagggtta gtaaaagggt cctaaggaac agcgatatct
     3541 cccaccccat gagctgtcac ggttttattt acatggggtc aggattccac gagggtagtg
     3601 aaccatttta gtcacaaggg cagtggctga agatcaagga gcgggcagtg aactctcctg
     3661 aatcttcgcc tgcttcttca ttctccttcg tttagctaat agaataactg ctgagttgtg
     3721 aacagtaagg tgtatgtgag gtgctcgaaa acaaggtttc aggtgacgcc cccagaataa
     3781 aatttggacg gggggttcag tggtggcatt gtgctatgac accaatataa ccctcacaaa
     3841 ccccttgggc aataaatact agtgtaggaa tgaaacattc tgaatatctt taacaataga
     3901 aatccatggg gtggggacaa gccgtaaaga ctggatgtcc atctcacacg aatttatggc
     3961 tatgggcaac acataatcct agtgcaatat gatactgggg ttattaagat gtgtcccagg
     4021 cagggaccaa gacaggtgaa ccatgttgtt acactctatt tgtaacaagg ggaaagagag
     4081 tggacgccga cagcagcgga ctccactggt tgtctctaac acccccgaaa attaaacggg
     4141 gctccacgcc aatggggccc ataaacaaag acaagtggcc actctttttt ttgaaattgt
     4201 ggagtggggg cacgcgtcag cccccacacg ccgccctgcg gttttggact gtaaaataag
     4261 ggtgtaataa cttggctgat tgtaaccccg ctaaccactg cggtcaaacc acttgcccac
     4321 aaaaccacta atggcacccc ggggaatacc tgcataagta ggtgggcggg ccaagatagg
     4381 ggcgcgattg ctgcgatctg gaggacaaat tacacacact tgcgcctgag cgccaagcac
     4441 agggttgttg gtcctcatat tcacgaggtc gctgagagca cggtgggcta atgttgccat
     4501 gggtagcata tactacccaa atatctggat agcatatgct atcctaatct atatctgggt
     4561 agcataggct atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt
     4621 agtatatgct atcctaattt atatctgggt agcataggct atcctaatct atatctgggt
     4681 agcatatgct atcctaatct atatctgggt agtatatgct atcctaatct gtatccgggt
     4741 agcatatgct atcctaatag agattagggt agtatatgct atcctaattt atatctgggt
     4801 agcatatact acccaaatat ctggatagca tatgctatcc taatctatat ctgggtagca
     4861 tatgctatcc taatctatat ctgggtagca taggctatcc taatctatat ctgggtagca
     4921 tatgctatcc taatctatat ctgggtagta tatgctatcc taatttatat ctgggtagca
     4981 taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagta
     5041 tatgctatcc taatctgtat ccgggtagca tatgctatcc tcatgataag ctgtcaaaca
     5101 tgagaatttt cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     5161 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     5221 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     5281 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     5341 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     5401 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     5461 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     5521 acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa
     5581 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     5641 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     5701 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     5761 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     5821 gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg
     5881 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     5941 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     6001 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     6061 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     6121 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     6181 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     6241 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     6301 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     6361 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     6421 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     6481 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     6541 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     6601 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     6661 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     6721 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     6781 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     6841 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     6901 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     6961 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     7021 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     7081 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     7141 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     7201 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     7261 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     7321 acaggaaaca gctatgacca tgattacgcc aagctctagc tagaggtcga ccaattctca
     7381 tgtttgacag cttatcatcg cagatccggg caacgttgtt gccattgctg caggcgcaga
     7441 actggtaggt atggaagatc tatacattga atcaatattg gcaattagcc atattagtca
     7501 ttggttatat agcataaatc aatattggct attggccatt gcatacgttg tatctatatc
     7561 ataatatgta catttatatt ggctcatgtc caatatgacc gc
//
