LOCUS       sequence-262352-        7383 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 7383)
  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 7383)
  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: D60_Ld_343410
FEATURES             Location/Qualifiers
     source          1..7383
                     /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             2547..2564
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2616..2671
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2670..2689)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4907..4925)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4993..5097
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5098..5166,5167..5958)
                     /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(5316..5335)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6129..6717
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6618..6637
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6871..6888
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7005..7026
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7041..7046,7047..7064,7065..7071)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7079..7095
                     /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     7084..7106
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7103..7119
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7103..7119
                     /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 gcggccgccc
     1201 atcaggtggc cctgatcagc gacatccact acgaccccct gtacggcacc agcagagccc
     1261 tgaagtgcac cagcgacagc agccctatgt acggcatgca gggctgcgac agctccctgc
     1321 agctgatggc cagaaccctg gccgacgtgt ccgcccagaa ccccagcctg gtgctgtaca
     1381 ccggcgactg gcagcggcac aagttcgcca acagctctct ggcccctgcc gccatcttca
     1441 aggacatgag cgagcggttc cacaacgtga ccgtggacgg ctctctgggc gccgtggcct
     1501 tttctggcgc catgggcaac aacgacgtgg tgcccgacta ctactacagc ctggaagata
     1561 ccgtgtccga ggaagaactg gcccaccggg tggccgccat gagaggcgct gctctgctgt
     1621 ctgatgccga ggccagcgtg atgcggaact gcggctacta cacccacatg atggacagcg
     1681 tgcacgtgat cgtgctgcac accctgctgt gggcccacga gctgcagccc cccctgccca
     1741 gcaatctgag cgacccctgc aaccagttca gcttcctgcg gagcgagctg gccaaagtgc
     1801 gggctgccaa caagcgggcc atcatcatgg gccacatccc ccctggcctg aacacctaca
     1861 acgtgctgaa gcggggcttc ggcagcgccg ctggcgacat gttctggaaa gagcagtacg
     1921 aggccaccta caacagcatc atcagagagt acaaggacct gctggtggtg cagctgttcg
     1981 gccacacaca catgttcaag ctgctgacca tgccccggaa cgacgtgctg ggcatcgtgg
     2041 tgcccagcat cagccccatc ttcggcaacc accccagcta cctgatggcc aacttcagcg
     2101 agacatggga gctggaagat gcccagagcc ggtacaccat cggcgagggc gtgttccaca
     2161 gcggcctgag cgccaaagag gtgctgagcc tgaacgccgg cctgcacagc gtgtcagacg
     2221 tgcgcgctgc catcaccctg ctggccacca acgataccat gtgggaccgg tttctgaccg
     2281 tgttctgtgg cggcgagaag cggagccagg tgttccccca ccggaagtgc gacaagcagt
     2341 gtcggtacat catcgtgtgc agcatgctgg aaaacaacca cacctacatc cagagatgcg
     2401 tggccaacta ctccctgctg cctggcccca gccaggacac cagaaccaca ggcgcgcccg
     2461 gcagcctgca ccacatcctg gacgcccaga agatgctgtg gaaccacagg gacaggaacc
     2521 tgcctcctct ggcccctctg ggcccccacc accatcacca ccattgagtc tagacacgtg
     2581 attaattaag gatcccccga cctcgacctc tggctaataa aggaaattta ttttcattgc
     2641 aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca
     2701 tttggtcgag atccctcgga gatctctagc tagaggatcg atccccgccc cggacgaact
     2761 aaacctgact acgacatctc tgccccttct tcgcggggca gtgcatgtaa tcccttcagt
     2821 tggttggtac aacttgccaa ctgggccctg ttccacatgt gacacggggg gggaccaaac
     2881 acaaaggggt tctctgactg tagttgacat ccttataaat ggatgtgcac atttgccaac
     2941 actgagtggc tttcatcctg gagcagactt tgcagtctgt ggactgcaac acaacattgc
     3001 ctttatgtgt aactcttggc tgaagctctt acaccaatgc tgggggacat gtacctccca
     3061 ggggcccagg aagactacgg gaggctacac caacgtcaat cagaggggcc tgtgtagcta
     3121 ccgataagcg gaccctcaag agggcattag caatagtgtt tataaggccc ccttgttaac
     3181 cctaaacggg tagcatatgc ttcccgggta gtagtatata ctatccagac taaccctaat
     3241 tcaatagcat atgttaccca acgggaagca tatgctatcg aattagggtt agtaaaaggg
     3301 tcctaaggaa cagcgatatc tcccacccca tgagctgtca cggttttatt tacatggggt
     3361 caggattcca cgagggtagt gaaccatttt agtcacaagg gcagtggctg aagatcaagg
     3421 agcgggcagt gaactctcct gaatcttcgc ctgcttcttc attctccttc gtttagctaa
     3481 tagaataact gctgagttgt gaacagtaag gtgtatgtga ggtgctcgaa aacaaggttt
     3541 caggtgacgc ccccagaata aaatttggac ggggggttca gtggtggcat tgtgctatga
     3601 caccaatata accctcacaa accccttggg caataaatac tagtgtagga atgaaacatt
     3661 ctgaatatct ttaacaatag aaatccatgg ggtggggaca agccgtaaag actggatgtc
     3721 catctcacac gaatttatgg ctatgggcaa cacataatcc tagtgcaata tgatactggg
     3781 gttattaaga tgtgtcccag gcagggacca agacaggtga accatgttgt tacactctat
     3841 ttgtaacaag gggaaagaga gtggacgccg acagcagcgg actccactgg ttgtctctaa
     3901 cacccccgaa aattaaacgg ggctccacgc caatggggcc cataaacaaa gacaagtggc
     3961 cactcttttt tttgaaattg tggagtgggg gcacgcgtca gcccccacac gccgccctgc
     4021 ggttttggac tgtaaaataa gggtgtaata acttggctga ttgtaacccc gctaaccact
     4081 gcggtcaaac cacttgccca caaaaccact aatggcaccc cggggaatac ctgcataagt
     4141 aggtgggcgg gccaagatag gggcgcgatt gctgcgatct ggaggacaaa ttacacacac
     4201 ttgcgcctga gcgccaagca cagggttgtt ggtcctcata ttcacgaggt cgctgagagc
     4261 acggtgggct aatgttgcca tgggtagcat atactaccca aatatctgga tagcatatgc
     4321 tatcctaatc tatatctggg tagcataggc tatcctaatc tatatctggg tagcatatgc
     4381 tatcctaatc tatatctggg tagtatatgc tatcctaatt tatatctggg tagcataggc
     4441 tatcctaatc tatatctggg tagcatatgc tatcctaatc tatatctggg tagtatatgc
     4501 tatcctaatc tgtatccggg tagcatatgc tatcctaata gagattaggg tagtatatgc
     4561 tatcctaatt tatatctggg tagcatatac tacccaaata tctggatagc atatgctatc
     4621 ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagc ataggctatc
     4681 ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt atatgctatc
     4741 ctaatttata tctgggtagc ataggctatc ctaatctata tctgggtagc atatgctatc
     4801 ctaatctata tctgggtagt atatgctatc ctaatctgta tccgggtagc atatgctatc
     4861 ctcatgataa gctgtcaaac atgagaattt tcttgaagac gaaagggcct cgtgatacgc
     4921 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     4981 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     5041 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     5101 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     5161 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     5221 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     5281 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     5341 gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     5401 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5461 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     5521 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     5581 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5641 gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5701 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     5761 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     5821 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     5881 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     5941 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     6001 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     6061 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     6121 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6181 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     6241 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     6301 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6361 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6421 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6481 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6541 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6601 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6661 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6721 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     6781 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     6841 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     6901 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     6961 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     7021 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     7081 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagctctag
     7141 ctagaggtcg accaattctc atgtttgaca gcttatcatc gcagatccgg gcaacgttgt
     7201 tgccattgct gcaggcgcag aactggtagg tatggaagat ctatacattg aatcaatatt
     7261 ggcaattagc catattagtc attggttata tagcataaat caatattggc tattggccat
     7321 tgcatacgtt gtatctatat cataatatgt acatttatat tggctcatgt ccaatatgac
     7381 cgc
//
