LOCUS       sequence-274860-        7251 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 7251)
  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 7251)
  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: D70_Ld_210610
FEATURES             Location/Qualifiers
     source          1..7251
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     201..225
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     706..725
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      748..757
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2029..2046
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2098..2153
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2152..2171)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4389..4407)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4475..4579
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4580..4648,4649..5440)
                     /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(4798..4817)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5611..6199
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6100..6119
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6353..6370
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6487..6508
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6523..6528,6529..6546,6547..6553)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6561..6577
                     /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     6566..6588
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6585..6601
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6585..6601
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        6872..7251
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta caccaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcctcac tctcttccgc
      241 atcgctgtct gcgagggcca gctgttgggc tcgcggttga ggacaaactc ttcgcggtct
      301 ttccagtact cttggatcgg aaacccgtcg gcctccgaac ggtactccgc caccgaggga
      361 cctgagcgag tccgcatcga ccggatcgga aaacctctcg agaaaggcgt ctaaccagtc
      421 acagtcgcaa ggtaggctga gcaccgtggc gggcggcagc gggtggcggt cggggttgtt
      481 tctggcggag gtgctgctga tgatgtaatt aaagtaggcg gtcttgagac ggcggatggt
      541 cgaggtgagg tgtggcaggc ttgagatcca gctgttgggg tgagtactcc ctctcaaaag
      601 cgggcattac ttctgcgcta agattgtcag tttccaaaaa cgaggaggat ttgatattca
      661 cctggcccga tctggccata cacttgagtg acaatgacat ccactttgcc tttctctcca
      721 caggtgtcca ctcccaggtc caagtttgcc accatggagt ttcagaccca ggtactcatg
      781 tccctgctgc tctgcatgtc tggtgcggcc gcccccagca tggaaaccgg aactgttgtg
      841 gaatggaaga agaaggtcgg agagcttgtg aaagaaagcg aagttttctg cacaattcag
      901 actgacaaag ccgtcgtgga ttacactaat acctttgaat caggctactt ggcgaagata
      961 tattgtggga acggacagag cgctccagtt gctaaaacta ttgccgttat ggtgtcagac
     1021 gccgcagatg tctctaaagc agacgaatat acacctgaag gcgaggtgcc cgcagctgat
     1081 gctgaggcac cagcagctgc ggcggtggca gcagctccag ctgccggcgg agcttcatct
     1141 gaagtgccgg aaggagtcac ctgtgaacca gtatttatgc ctgcactctc ccccagtatg
     1201 gagacaggca cagtcgtgga gtggaagaag aaagtcggag aactcgtgaa agagtctgag
     1261 gtgttttgta ccattcaaac ggacaaagcc gtagttgatt acaccaacac gtttgaatct
     1321 ggttatcttg ctaagatcta ctgtgggaat ggtcagtccg caccagtggc taagacaatt
     1381 gccgtgatgg tttccaatgc cgccgatgtc gagaaagtgg ctaattatta tcctgaggat
     1441 gcagtaggcg gaccaccggc atctgccgct gacccaagcg ctgcagcagc agccgcagca
     1501 tctgcaaggc cagctgcttc agctgccagt gcgaaacact atggcgggtc tttggacgcc
     1561 gccgtggccg caagtggacc atctgtggcc agaattgctg ctggcctcga accaagtgcc
     1621 cttgctggga ttgctcctag tgggaaaggc gggagattct tgaagagtga tttcagcggc
     1681 caacccggat tcgactacaa cgatactgcg cccgctcgcg ctatgcaaca aaaggctgct
     1741 cccgcagctg ccgccgatgg cgcttcaaag gccgctgcaa aatcagctgc acccgcggct
     1801 gtcagcggag atatttacaa cgtggttctt aagcccggcc ccgtttataa aagcgtctca
     1861 gataccgcac tgctgaagaa gctgatgtgt tccatgcaca tcccaaaacc gaaaagcaag
     1921 aaggcagccg aaggcgcgcc cggcagcctg caccacatcc tggacgccca gaagatgctg
     1981 tggaaccaca gggacaggaa cctgcctcct ctggcccctc tgggccccca ccaccatcac
     2041 caccattgag tctagacacg tgattaatta aggatccccc gacctcgacc tctggctaat
     2101 aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa
     2161 ggacatatgg gagggcaaat catttggtcg agatccctcg gagatctcta gctagaggat
     2221 cgatccccgc cccggacgaa ctaaacctga ctacgacatc tctgcccctt cttcgcgggg
     2281 cagtgcatgt aatcccttca gttggttggt acaacttgcc aactgggccc tgttccacat
     2341 gtgacacggg gggggaccaa acacaaaggg gttctctgac tgtagttgac atccttataa
     2401 atggatgtgc acatttgcca acactgagtg gctttcatcc tggagcagac tttgcagtct
     2461 gtggactgca acacaacatt gcctttatgt gtaactcttg gctgaagctc ttacaccaat
     2521 gctgggggac atgtacctcc caggggccca ggaagactac gggaggctac accaacgtca
     2581 atcagagggg cctgtgtagc taccgataag cggaccctca agagggcatt agcaatagtg
     2641 tttataaggc ccccttgtta accctaaacg ggtagcatat gcttcccggg tagtagtata
     2701 tactatccag actaacccta attcaatagc atatgttacc caacgggaag catatgctat
     2761 cgaattaggg ttagtaaaag ggtcctaagg aacagcgata tctcccaccc catgagctgt
     2821 cacggtttta tttacatggg gtcaggattc cacgagggta gtgaaccatt ttagtcacaa
     2881 gggcagtggc tgaagatcaa ggagcgggca gtgaactctc ctgaatcttc gcctgcttct
     2941 tcattctcct tcgtttagct aatagaataa ctgctgagtt gtgaacagta aggtgtatgt
     3001 gaggtgctcg aaaacaaggt ttcaggtgac gcccccagaa taaaatttgg acggggggtt
     3061 cagtggtggc attgtgctat gacaccaata taaccctcac aaaccccttg ggcaataaat
     3121 actagtgtag gaatgaaaca ttctgaatat ctttaacaat agaaatccat ggggtgggga
     3181 caagccgtaa agactggatg tccatctcac acgaatttat ggctatgggc aacacataat
     3241 cctagtgcaa tatgatactg gggttattaa gatgtgtccc aggcagggac caagacaggt
     3301 gaaccatgtt gttacactct atttgtaaca aggggaaaga gagtggacgc cgacagcagc
     3361 ggactccact ggttgtctct aacacccccg aaaattaaac ggggctccac gccaatgggg
     3421 cccataaaca aagacaagtg gccactcttt tttttgaaat tgtggagtgg gggcacgcgt
     3481 cagcccccac acgccgccct gcggttttgg actgtaaaat aagggtgtaa taacttggct
     3541 gattgtaacc ccgctaacca ctgcggtcaa accacttgcc cacaaaacca ctaatggcac
     3601 cccggggaat acctgcataa gtaggtgggc gggccaagat aggggcgcga ttgctgcgat
     3661 ctggaggaca aattacacac acttgcgcct gagcgccaag cacagggttg ttggtcctca
     3721 tattcacgag gtcgctgaga gcacggtggg ctaatgttgc catgggtagc atatactacc
     3781 caaatatctg gatagcatat gctatcctaa tctatatctg ggtagcatag gctatcctaa
     3841 tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagtatat gctatcctaa
     3901 tttatatctg ggtagcatag gctatcctaa tctatatctg ggtagcatat gctatcctaa
     3961 tctatatctg ggtagtatat gctatcctaa tctgtatccg ggtagcatat gctatcctaa
     4021 tagagattag ggtagtatat gctatcctaa tttatatctg ggtagcatat actacccaaa
     4081 tatctggata gcatatgcta tcctaatcta tatctgggta gcatatgcta tcctaatcta
     4141 tatctgggta gcataggcta tcctaatcta tatctgggta gcatatgcta tcctaatcta
     4201 tatctgggta gtatatgcta tcctaattta tatctgggta gcataggcta tcctaatcta
     4261 tatctgggta gcatatgcta tcctaatcta tatctgggta gtatatgcta tcctaatctg
     4321 tatccgggta gcatatgcta tcctcatgat aagctgtcaa acatgagaat tttcttgaag
     4381 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     4441 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     4501 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     4561 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     4621 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     4681 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     4741 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     4801 atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca
     4861 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     4921 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     4981 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     5041 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     5101 cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg
     5161 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     5221 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     5281 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     5341 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     5401 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     5461 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     5521 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     5581 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5641 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5701 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     5761 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5821 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5881 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     5941 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     6001 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     6061 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     6121 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6181 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6241 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     6301 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     6361 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     6421 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     6481 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     6541 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     6601 ccatgattac gccaagctct agctagaggt cgaccaattc tcatgtttga cagcttatca
     6661 tcgcagatcc gggcaacgtt gttgccattg ctgcaggcgc agaactggta ggtatggaag
     6721 atctatacat tgaatcaata ttggcaatta gccatattag tcattggtta tatagcataa
     6781 atcaatattg gctattggcc attgcatacg ttgtatctat atcataatat gtacatttat
     6841 attggctcat gtccaatatg accgccatgt tgacattgat tattgactag ttattaatag
     6901 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
     6961 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     7021 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     7081 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct
     7141 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg
     7201 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat g
//
