LOCUS       sequence-262350-        7755 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 7755)
  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 7755)
  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: D58_Ld_280280
FEATURES             Location/Qualifiers
     source          1..7755
                     /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             2919..2936
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2988..3043
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3042..3061)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5279..5297)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5365..5469
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5470..5538,5539..6330)
                     /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(5688..5707)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6501..7089
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6990..7009
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7243..7260
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7377..7398
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7413..7418,7419..7436,7437..7443)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7451..7467
                     /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     7456..7478
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7475..7491
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7475..7491
                     /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 tggatgagca gcacagctac tacgagggca actacggcac caccggcctg gtgttcggca
     1261 gccccgatag cagccggacc agctgggact acaccaacct gaaggactgg cccaccctgt
     1321 gcctgaccgg cagaaggcag agccccatca gcttcgccaa cgtgaacccc gacgaggtgg
     1381 tcaccaacgc cctgctgcag cggctgcagt tcagctctaa gtgcgtgttc ccccgggaag
     1441 agacacagat gcggatcatc aacgagggca tcgtgtccac cgtgtccttc gaggaactgg
     1501 gccggtccct ggacgacatg agcgagtgca ccgtgatgga ccccctgaac agaagcctga
     1561 cctaccactt catgggcctg cacttccacg ccatgcccgt gcaccagctg agaaccctga
     1621 ggcccgacgc cgagatgcac atgagcttca ccaccaacca cgtggaacag aaaatgcgga
     1681 acgtgctgat cgtggccgtg atgctgaagg ccagcgccat ggtcaacagc accagcgcca
     1741 gagccctgca gcacatcctg gtggacggca gcctgcccaa gcggaacgcc atgaccacct
     1801 gttttctgac cgagagcctg agcctgttca gcctgatccc cgccagagag agctacctgc
     1861 tgtacgacgg cagccagacc caccccccct gcaccgagaa cgtccgctgg gtcgtgatga
     1921 ccagccccat cctgatcagc cccgtggccc tgggcagact gagagatgcc atggatgccc
     1981 tgctgcccaa cgacttccac agattcggca acgccagacc cccccaggcc ctgaacggcc
     2041 ggctgatctt cagattcgac gacagaagcg tgcccgacgg cggcaaccgg ggcgagggca
     2101 atttcaagga cgcctggtcc cacaaggacg agggctgggc caacagatcc ctgggcgtca
     2161 gagccggcag agtgctggaa ggcagcctgg tggaagtgga cggcgactat aagtacgaca
     2221 gcgccgtggt ggtgtcccct gtgtccccca atcccgtgtc cgccagccgg aatggcacca
     2281 cccacgccag cagcccccct atcaccacca gccccggcag atacaacggc agcagcgacc
     2341 aggccatggc caagcctgga acaggcgccc tgggccctgc ctacatggcc aacgccagca
     2401 gaaccggccc tgagacacac agccacgtga acgagagcga cgccgactac agcaacgcca
     2461 ccgccaatgc cagctccgcc cccagaacca gcgtggtgtc tagcgccagc gccagcagcg
     2521 ccgacgagag catgccttct ggcctgcacc acggcagcga gagcagcgcc cctacccctt
     2581 ctgacgagag cgcccccaga tccaccacca ccagcacaac caccacaact accaccacca
     2641 aagagcccag cggcaccaag ggcaagaaga gatacggcga cggcgccagc aagaacacca
     2701 gcgaagccga cgccggcgtg gtggacagcg tgaagtccat cgtgacccgg gccttcacca
     2761 gcgtgaaggc cttctccatc ggcgccttcc agagcagcgt ggcctacgcc aaggccaacc
     2821 ctggcgcgcc cggcagcctg caccacatcc tggacgccca gaagatgctg tggaaccaca
     2881 gggacaggaa cctgcctcct ctggcccctc tgggccccca ccaccatcac caccattgag
     2941 tctagacacg tgattaatta aggatccccc gacctcgacc tctggctaat aaaggaaatt
     3001 tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg
     3061 gagggcaaat catttggtcg agatccctcg gagatctcta gctagaggat cgatccccgc
     3121 cccggacgaa ctaaacctga ctacgacatc tctgcccctt cttcgcgggg cagtgcatgt
     3181 aatcccttca gttggttggt acaacttgcc aactgggccc tgttccacat gtgacacggg
     3241 gggggaccaa acacaaaggg gttctctgac tgtagttgac atccttataa atggatgtgc
     3301 acatttgcca acactgagtg gctttcatcc tggagcagac tttgcagtct gtggactgca
     3361 acacaacatt gcctttatgt gtaactcttg gctgaagctc ttacaccaat gctgggggac
     3421 atgtacctcc caggggccca ggaagactac gggaggctac accaacgtca atcagagggg
     3481 cctgtgtagc taccgataag cggaccctca agagggcatt agcaatagtg tttataaggc
     3541 ccccttgtta accctaaacg ggtagcatat gcttcccggg tagtagtata tactatccag
     3601 actaacccta attcaatagc atatgttacc caacgggaag catatgctat cgaattaggg
     3661 ttagtaaaag ggtcctaagg aacagcgata tctcccaccc catgagctgt cacggtttta
     3721 tttacatggg gtcaggattc cacgagggta gtgaaccatt ttagtcacaa gggcagtggc
     3781 tgaagatcaa ggagcgggca gtgaactctc ctgaatcttc gcctgcttct tcattctcct
     3841 tcgtttagct aatagaataa ctgctgagtt gtgaacagta aggtgtatgt gaggtgctcg
     3901 aaaacaaggt ttcaggtgac gcccccagaa taaaatttgg acggggggtt cagtggtggc
     3961 attgtgctat gacaccaata taaccctcac aaaccccttg ggcaataaat actagtgtag
     4021 gaatgaaaca ttctgaatat ctttaacaat agaaatccat ggggtgggga caagccgtaa
     4081 agactggatg tccatctcac acgaatttat ggctatgggc aacacataat cctagtgcaa
     4141 tatgatactg gggttattaa gatgtgtccc aggcagggac caagacaggt gaaccatgtt
     4201 gttacactct atttgtaaca aggggaaaga gagtggacgc cgacagcagc ggactccact
     4261 ggttgtctct aacacccccg aaaattaaac ggggctccac gccaatgggg cccataaaca
     4321 aagacaagtg gccactcttt tttttgaaat tgtggagtgg gggcacgcgt cagcccccac
     4381 acgccgccct gcggttttgg actgtaaaat aagggtgtaa taacttggct gattgtaacc
     4441 ccgctaacca ctgcggtcaa accacttgcc cacaaaacca ctaatggcac cccggggaat
     4501 acctgcataa gtaggtgggc gggccaagat aggggcgcga ttgctgcgat ctggaggaca
     4561 aattacacac acttgcgcct gagcgccaag cacagggttg ttggtcctca tattcacgag
     4621 gtcgctgaga gcacggtggg ctaatgttgc catgggtagc atatactacc caaatatctg
     4681 gatagcatat gctatcctaa tctatatctg ggtagcatag gctatcctaa tctatatctg
     4741 ggtagcatat gctatcctaa tctatatctg ggtagtatat gctatcctaa tttatatctg
     4801 ggtagcatag gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg
     4861 ggtagtatat gctatcctaa tctgtatccg ggtagcatat gctatcctaa tagagattag
     4921 ggtagtatat gctatcctaa tttatatctg ggtagcatat actacccaaa tatctggata
     4981 gcatatgcta tcctaatcta tatctgggta gcatatgcta tcctaatcta tatctgggta
     5041 gcataggcta tcctaatcta tatctgggta gcatatgcta tcctaatcta tatctgggta
     5101 gtatatgcta tcctaattta tatctgggta gcataggcta tcctaatcta tatctgggta
     5161 gcatatgcta tcctaatcta tatctgggta gtatatgcta tcctaatctg tatccgggta
     5221 gcatatgcta tcctcatgat aagctgtcaa acatgagaat tttcttgaag acgaaagggc
     5281 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     5341 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     5401 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     5461 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     5521 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     5581 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     5641 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     5701 gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     5761 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     5821 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     5881 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     5941 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     6001 accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     6061 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     6121 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     6181 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     6241 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     6301 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     6361 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     6421 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     6481 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     6541 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     6601 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     6661 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     6721 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     6781 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     6841 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     6901 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6961 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     7021 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     7081 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     7141 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     7201 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     7261 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     7321 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     7381 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     7441 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     7501 gccaagctct agctagaggt cgaccaattc tcatgtttga cagcttatca tcgcagatcc
     7561 gggcaacgtt gttgccattg ctgcaggcgc agaactggta ggtatggaag atctatacat
     7621 tgaatcaata ttggcaatta gccatattag tcattggtta tatagcataa atcaatattg
     7681 gctattggcc attgcatacg ttgtatctat atcataatat gtacatttat attggctcat
     7741 gtccaatatg accgc
//
