LOCUS       sequence-262318-        8643 bp    DNA     circular SYN 15-APR-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 8643)
  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 8643)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: D23_Ld_170240
FEATURES             Location/Qualifiers
     source          1..8643
                     /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             3807..3824
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3876..3931
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3930..3949)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6167..6185)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6253..6357
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6358..6426,6427..7218)
                     /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(6576..6595)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7389..7977
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7878..7897
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8131..8148
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8265..8286
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8301..8306,8307..8324,8325..8331)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8339..8355
                     /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     8344..8366
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8363..8379
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8363..8379
                     /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 gcggccgcca
     1201 cctttgatgg cgacgccgtg acagcccctc ctagcgccag caccgccttc accgagcacc
     1261 ctttcaccct gcctaccgcc gccagcctgc tggacgtgct ggccagcgac ttctgcacca
     1321 gaaccggcga gggcgaggaa agcagctgca cccccagcgt gtccagactg cccacccacc
     1381 ccgtgctgat cctgtttacc gagagcaacg ccggccacag cagcaccctg gccgccctga
     1441 gagccaaagt gcggcagatc gccaccgacg tgcccagcag cgtgctgcgg gtgcacgagt
     1501 acgtgatccc catgagcgcc acagagcggc ggctgatcga gctgctgctg atcggcgcca
     1561 gccacgtgcc aacactggcc ctgtttcacg gccgggtggc caaggtgcag gtcgcccctg
     1621 gatccatcgt gtctgccagc cccttccaga cccctgtgct gtactccacc acccccctgg
     1681 cctccgccag ctacctggaa ctgagaagct gggccctgtc cgagctgccc gccagatacg
     1741 tggaccccgt gaccttccac ctgatcccta gcctgcagtt cgtgttcagc cctgccgaag
     1801 tgcacgaaac actgcagctc gtgcgggacg ccctggaagc cagcgagagc agatccgtgc
     1861 tgcctgcctt cgtgtccatg gcctacatgc ggctgacccg gcacggcagc gaagaggtgc
     1921 tggctgccct gtctagcatt gccacccagg ccggcaacgc tgtgctgacc ctggtcaccg
     1981 aaagcgccga ggtggccgct gcctggggac tgagagctga gcacaccttc accatggccc
     2041 cttggtccag cacagtggcc gcctacctga acagcagcgc ctccgatggc ttcgccgcct
     2101 ccgtgtccga tctgagccct ccccagagca tcggcaccgt gaccgagctg gccgaagcca
     2161 cagccggctc tgccgcttgg agagccgctt gtgccgcctc tgtgcgggcc gagatcgccc
     2221 agctgcagaa ctggcggcag gccatgaacg ccttcaacac caccagccct ctgcggcgga
     2281 tcgacagcgc cgctcatctg ctgcacgagc tgaaggccct gcagaatgcc ctgaagatcg
     2341 tgttcgtgct gagagaaagc gacgagatgt ggtttcacca ccacctggaa gtggctgccg
     2401 ctctggccag aaggctgcag agcaccaccg tgttctacaa cacaaccaca ctggctaagg
     2461 gcagcaaggc ccggtcccgg gtggaaagat cctggacccc catcatgaga tgcgacgtgt
     2521 tctggctgga cgccgagcag ctccctgccg tggctgatgg actgcacgtg tcccaggtgc
     2581 ccagtgtgct gattctggtg cccctgcagt ccagattcgg agagggcggc gacggcgata
     2641 gagccgatgc tgctgcagcc gctggcgacg acgatcaggc agctgccggc gcttctgagg
     2701 aagccggcct gagaaggcgg gaccccttca tcggcatcca caccatcaac agatacgacc
     2761 tgctgaaagc cgcctatatg gacgacgccc agatgaccca cgagcctgtg tccggcaaag
     2821 aggcactgcc tctgttcccc agcgattccg atgccctgct gcggttcctg gctagcggca
     2881 gctttctggg cgccatgcag tacaccatgt cctccaccca gctgtctacc ctgcgggcta
     2941 aagtgacaga gctgcctgat gccgtgcagg ccgctaccgc cagcggcgga accagcaaca
     3001 ggcactacct gcagctggac cggcggtact accctctgag aggggccgag gaacccatgg
     3061 aaggccccgt gtacgtcaga cagatcctga acggctccag ccccctgcca gtgctgggct
     3121 ccgacgacag cgacaccgaa ctgggcagac agcccgtggg cggcagaagc ggcacacacc
     3181 ctagaggcac cacctccgcc tcccccgacg aggccgatga gagactgaga cagcggagag
     3241 ccagagaaga ggccgcagcc ctgaaacgga aggccgcctg ggaggccaat ctggccaagc
     3301 ggagaaggga aaaggccgac cggatgcgga gaaaggctgc cgaggatgcc gccgagcgcg
     3361 agaggcagca ggaagccttc cagcaggaag tgaacgccgc cctgaaggcc gaacaggaag
     3421 gcgctggcgg cggagctggc ggaacatggg cttctcagcc tgagggcctg ctcgtcagaa
     3481 ggcccactct ggaagatgcc agcgaaggcc tggtggaaga gggcagacgg aagggcaggc
     3541 cccagaggcc cttcggggag gaagtggata gccaggacgc tgcccagatt ctccggcgga
     3601 ggcggagaca ccgcgagtac agagagtggc tggctgacag acagctgatg gtcaatagat
     3661 gcgtgacagt gaccgaaaag ggcaagctga gcctgatgac cagatgggaa ggcgcgcccg
     3721 gcagcctgca ccacatcctg gacgcccaga agatgctgtg gaaccacagg gacaggaacc
     3781 tgcctcctct ggcccctctg ggcccccacc accatcacca ccattgagtc tagacacgtg
     3841 attaattaag gatcccccga cctcgacctc tggctaataa aggaaattta ttttcattgc
     3901 aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca
     3961 tttggtcgag atccctcgga gatctctagc tagaggatcg atccccgccc cggacgaact
     4021 aaacctgact acgacatctc tgccccttct tcgcggggca gtgcatgtaa tcccttcagt
     4081 tggttggtac aacttgccaa ctgggccctg ttccacatgt gacacggggg gggaccaaac
     4141 acaaaggggt tctctgactg tagttgacat ccttataaat ggatgtgcac atttgccaac
     4201 actgagtggc tttcatcctg gagcagactt tgcagtctgt ggactgcaac acaacattgc
     4261 ctttatgtgt aactcttggc tgaagctctt acaccaatgc tgggggacat gtacctccca
     4321 ggggcccagg aagactacgg gaggctacac caacgtcaat cagaggggcc tgtgtagcta
     4381 ccgataagcg gaccctcaag agggcattag caatagtgtt tataaggccc ccttgttaac
     4441 cctaaacggg tagcatatgc ttcccgggta gtagtatata ctatccagac taaccctaat
     4501 tcaatagcat atgttaccca acgggaagca tatgctatcg aattagggtt agtaaaaggg
     4561 tcctaaggaa cagcgatatc tcccacccca tgagctgtca cggttttatt tacatggggt
     4621 caggattcca cgagggtagt gaaccatttt agtcacaagg gcagtggctg aagatcaagg
     4681 agcgggcagt gaactctcct gaatcttcgc ctgcttcttc attctccttc gtttagctaa
     4741 tagaataact gctgagttgt gaacagtaag gtgtatgtga ggtgctcgaa aacaaggttt
     4801 caggtgacgc ccccagaata aaatttggac ggggggttca gtggtggcat tgtgctatga
     4861 caccaatata accctcacaa accccttggg caataaatac tagtgtagga atgaaacatt
     4921 ctgaatatct ttaacaatag aaatccatgg ggtggggaca agccgtaaag actggatgtc
     4981 catctcacac gaatttatgg ctatgggcaa cacataatcc tagtgcaata tgatactggg
     5041 gttattaaga tgtgtcccag gcagggacca agacaggtga accatgttgt tacactctat
     5101 ttgtaacaag gggaaagaga gtggacgccg acagcagcgg actccactgg ttgtctctaa
     5161 cacccccgaa aattaaacgg ggctccacgc caatggggcc cataaacaaa gacaagtggc
     5221 cactcttttt tttgaaattg tggagtgggg gcacgcgtca gcccccacac gccgccctgc
     5281 ggttttggac tgtaaaataa gggtgtaata acttggctga ttgtaacccc gctaaccact
     5341 gcggtcaaac cacttgccca caaaaccact aatggcaccc cggggaatac ctgcataagt
     5401 aggtgggcgg gccaagatag gggcgcgatt gctgcgatct ggaggacaaa ttacacacac
     5461 ttgcgcctga gcgccaagca cagggttgtt ggtcctcata ttcacgaggt cgctgagagc
     5521 acggtgggct aatgttgcca tgggtagcat atactaccca aatatctgga tagcatatgc
     5581 tatcctaatc tatatctggg tagcataggc tatcctaatc tatatctggg tagcatatgc
     5641 tatcctaatc tatatctggg tagtatatgc tatcctaatt tatatctggg tagcataggc
     5701 tatcctaatc tatatctggg tagcatatgc tatcctaatc tatatctggg tagtatatgc
     5761 tatcctaatc tgtatccggg tagcatatgc tatcctaata gagattaggg tagtatatgc
     5821 tatcctaatt tatatctggg tagcatatac tacccaaata tctggatagc atatgctatc
     5881 ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagc ataggctatc
     5941 ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt atatgctatc
     6001 ctaatttata tctgggtagc ataggctatc ctaatctata tctgggtagc atatgctatc
     6061 ctaatctata tctgggtagt atatgctatc ctaatctgta tccgggtagc atatgctatc
     6121 ctcatgataa gctgtcaaac atgagaattt tcttgaagac gaaagggcct cgtgatacgc
     6181 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     6241 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     6301 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     6361 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     6421 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     6481 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     6541 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     6601 gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     6661 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     6721 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     6781 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     6841 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     6901 gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     6961 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     7021 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     7081 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     7141 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     7201 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     7261 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     7321 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     7381 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     7441 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     7501 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     7561 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     7621 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     7681 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     7741 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     7801 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     7861 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     7921 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     7981 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     8041 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     8101 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     8161 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     8221 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     8281 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     8341 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagctctag
     8401 ctagaggtcg accaattctc atgtttgaca gcttatcatc gcagatccgg gcaacgttgt
     8461 tgccattgct gcaggcgcag aactggtagg tatggaagat ctatacattg aatcaatatt
     8521 ggcaattagc catattagtc attggttata tagcataaat caatattggc tattggccat
     8581 tgcatacgtt gtatctatat cataatatgt acatttatat tggctcatgt ccaatatgac
     8641 cgc
//
