LOCUS       sequence-262309-        7575 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 7575)
  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 7575)
  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: D14_Ld_051070
FEATURES             Location/Qualifiers
     source          1..7575
                     /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             2739..2756
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2808..2863
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2862..2881)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5099..5117)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5185..5289
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5290..5358,5359..6150)
                     /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(5508..5527)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6321..6909
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6810..6829
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7063..7080
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7197..7218
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7233..7238,7239..7256,7257..7263)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7271..7287
                     /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     7276..7298
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7295..7311
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7295..7311
                     /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 ccgccagagt gaacgatgat ctgccccctg gcgtgatgag aggccccctg ctgcacgagc
     1261 acagcttcca ggaacccctc gtcagcgatt ggtgggagga aggcgtgccc cactacatga
     1321 tcggcggcag cgccgtggcc aacgagcggt tcctgagact gaccaccaac gacctggacg
     1381 accacggctt cgccttcaac accgcccctc tggaccacga caactgggag gctagagtgc
     1441 ggttcagcat cagacccccc ccactggccg ccgacatgaa cggctctagc cactatcagg
     1501 gcggcgacgg catggccctg tggtatctgg atcagcccat cggcgacgat caccagcacg
     1561 tggtcaagta cagcaagagc atctcccccg agctgcggga cgagatgctg gacgccgaga
     1621 acccttggcg ggtggccgac ttcctgctga atggcgacga cgatgacgac gaggatgagg
     1681 gcgtggacga ggacgacctg accgagcagg aaaagcagga acggcagcag gccaaggaca
     1741 gacgggccac caagaagcgg aagcgcgagg ccctgttcaa gcggctgttt aagcggggca
     1801 cctccgtgga tgagagcgac cgcgagcccc ggatcatggg cgtgaagttc agcgacttca
     1861 acaagggctt cggcctgatc ctggacagcg tgggcgacga gaaagagcac ctgagagagc
     1921 accagcagca gggcgacgcc gtgcagcacc accaccatac cagctccatc tacctgctgc
     1981 tgaacctgcc caaccacacc tccagctccg gcgccaccgt cgtgaacaac ttcgacccca
     2041 ccagagagaa cttccggtcc agcccccacg tgctgcggtg cacctacgac ttccggcaga
     2101 agggcaccaa gccctacgac cccaaggccc ggatgatgga caaagtgacc aaagccgcca
     2161 cagccgccga ggaacctctg gaactgatcg tgcggtacta caagcagaaa ctgaccgtga
     2221 tcatccggcg cgaggatgtg gccaagcggg aagtgtcctg ggtggtggac gaaatgggca
     2281 aggccaccga cgaggtggaa gtcgtgcgga cctacaccga gacaatgtgc ggcgaggtgt
     2341 accccatcca tctgcctctg agataccact ttggcctgag cgccagcacc ggccacagac
     2401 ccgccaagaa cagaaagaga aggcccctgc ggagcgtgtt ccacgaggcc gacaagaaaa
     2461 ccttcaccca cgtggacgtg cacgacgtgc tgaagttcga cctgcgcgag ctgggcctgg
     2521 atgccaaggc catgggctac tccaagacca tccccctgga acacttcgat tttgaggccg
     2581 atcggagaga gcggctgcac ttcagccggc agctgcccgt gaaccccgac agcgataccg
     2641 ctggcgcgcc cggcagcctg caccacatcc tggacgccca gaagatgctg tggaaccaca
     2701 gggacaggaa cctgcctcct ctggcccctc tgggccccca ccaccatcac caccattgag
     2761 tctagacacg tgattaatta aggatccccc gacctcgacc tctggctaat aaaggaaatt
     2821 tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg
     2881 gagggcaaat catttggtcg agatccctcg gagatctcta gctagaggat cgatccccgc
     2941 cccggacgaa ctaaacctga ctacgacatc tctgcccctt cttcgcgggg cagtgcatgt
     3001 aatcccttca gttggttggt acaacttgcc aactgggccc tgttccacat gtgacacggg
     3061 gggggaccaa acacaaaggg gttctctgac tgtagttgac atccttataa atggatgtgc
     3121 acatttgcca acactgagtg gctttcatcc tggagcagac tttgcagtct gtggactgca
     3181 acacaacatt gcctttatgt gtaactcttg gctgaagctc ttacaccaat gctgggggac
     3241 atgtacctcc caggggccca ggaagactac gggaggctac accaacgtca atcagagggg
     3301 cctgtgtagc taccgataag cggaccctca agagggcatt agcaatagtg tttataaggc
     3361 ccccttgtta accctaaacg ggtagcatat gcttcccggg tagtagtata tactatccag
     3421 actaacccta attcaatagc atatgttacc caacgggaag catatgctat cgaattaggg
     3481 ttagtaaaag ggtcctaagg aacagcgata tctcccaccc catgagctgt cacggtttta
     3541 tttacatggg gtcaggattc cacgagggta gtgaaccatt ttagtcacaa gggcagtggc
     3601 tgaagatcaa ggagcgggca gtgaactctc ctgaatcttc gcctgcttct tcattctcct
     3661 tcgtttagct aatagaataa ctgctgagtt gtgaacagta aggtgtatgt gaggtgctcg
     3721 aaaacaaggt ttcaggtgac gcccccagaa taaaatttgg acggggggtt cagtggtggc
     3781 attgtgctat gacaccaata taaccctcac aaaccccttg ggcaataaat actagtgtag
     3841 gaatgaaaca ttctgaatat ctttaacaat agaaatccat ggggtgggga caagccgtaa
     3901 agactggatg tccatctcac acgaatttat ggctatgggc aacacataat cctagtgcaa
     3961 tatgatactg gggttattaa gatgtgtccc aggcagggac caagacaggt gaaccatgtt
     4021 gttacactct atttgtaaca aggggaaaga gagtggacgc cgacagcagc ggactccact
     4081 ggttgtctct aacacccccg aaaattaaac ggggctccac gccaatgggg cccataaaca
     4141 aagacaagtg gccactcttt tttttgaaat tgtggagtgg gggcacgcgt cagcccccac
     4201 acgccgccct gcggttttgg actgtaaaat aagggtgtaa taacttggct gattgtaacc
     4261 ccgctaacca ctgcggtcaa accacttgcc cacaaaacca ctaatggcac cccggggaat
     4321 acctgcataa gtaggtgggc gggccaagat aggggcgcga ttgctgcgat ctggaggaca
     4381 aattacacac acttgcgcct gagcgccaag cacagggttg ttggtcctca tattcacgag
     4441 gtcgctgaga gcacggtggg ctaatgttgc catgggtagc atatactacc caaatatctg
     4501 gatagcatat gctatcctaa tctatatctg ggtagcatag gctatcctaa tctatatctg
     4561 ggtagcatat gctatcctaa tctatatctg ggtagtatat gctatcctaa tttatatctg
     4621 ggtagcatag gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg
     4681 ggtagtatat gctatcctaa tctgtatccg ggtagcatat gctatcctaa tagagattag
     4741 ggtagtatat gctatcctaa tttatatctg ggtagcatat actacccaaa tatctggata
     4801 gcatatgcta tcctaatcta tatctgggta gcatatgcta tcctaatcta tatctgggta
     4861 gcataggcta tcctaatcta tatctgggta gcatatgcta tcctaatcta tatctgggta
     4921 gtatatgcta tcctaattta tatctgggta gcataggcta tcctaatcta tatctgggta
     4981 gcatatgcta tcctaatcta tatctgggta gtatatgcta tcctaatctg tatccgggta
     5041 gcatatgcta tcctcatgat aagctgtcaa acatgagaat tttcttgaag acgaaagggc
     5101 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     5161 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     5221 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     5281 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     5341 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     5401 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     5461 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     5521 gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     5581 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     5641 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     5701 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     5761 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     5821 accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     5881 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     5941 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     6001 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     6061 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     6121 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     6181 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     6241 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     6301 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     6361 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     6421 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     6481 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     6541 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     6601 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     6661 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     6721 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6781 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6841 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6901 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6961 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     7021 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     7081 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     7141 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     7201 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     7261 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     7321 gccaagctct agctagaggt cgaccaattc tcatgtttga cagcttatca tcgcagatcc
     7381 gggcaacgtt gttgccattg ctgcaggcgc agaactggta ggtatggaag atctatacat
     7441 tgaatcaata ttggcaatta gccatattag tcattggtta tatagcataa atcaatattg
     7501 gctattggcc attgcatacg ttgtatctat atcataatat gtacatttat attggctcat
     7561 gtccaatatg accgc
//
