LOCUS       sequence-274546-        7221 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 7221)
  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 7221)
  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: D21_Ld_141150
FEATURES             Location/Qualifiers
     source          1..7221
                     /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             1999..2016
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2068..2123
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2122..2141)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4359..4377)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4445..4549
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4550..4618,4619..5410)
                     /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(4768..4787)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5581..6169
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6070..6089
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6323..6340
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6457..6478
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6493..6498,6499..6516,6517..6523)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6531..6547
                     /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     6536..6558
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6555..6571
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6555..6571
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        6842..7221
                     /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 gccaccagca gcttttgcct gagaggcaga
      841 gacacccacg gccccagact gagctgtgac gcccacctga cagtgtccgc cgtggccctg
      901 ttctactacg gcagcagcga gatcaacaga tgcgccgtgc agagcagagc caccgccgtg
      961 cctggcgtgc tgatggtccg agccgccaca gccgaaagca gcgctggcgg ctctgaggaa
     1021 gaagccggcg gaacagacgc ctggggcaca atcgccatga agcgggccct gaactacgac
     1081 agcggcctga agcccttcat gaccccctgg cgggaagtgc ggagagccct gagagatgcc
     1141 ggctgcttca gcaccctggc cctgcctaga ctgaccctgc ccaccgatag agccagcccc
     1201 agagtgggcg acgtgtcccc agaatccagc accgccctgc cccccctgtg cagaaagccc
     1261 ctggtcctga tgagcggcag aggcagggat tgcgccagct gcagctcctt cgccagcgac
     1321 gtgctggaac actaccgggc cgagtgggag tacatccggg cccacgtgac cctggtggac
     1381 gccggcgatc ccatgaccga ggaactcgtg cggaacccct accctctgtg gtacgatccc
     1441 gccgaggccg gaccctggat gaccctgacc aaggtgtccg aagtgcgcag ggctgagggg
     1501 gatgtggtgg ccgctgccag cggcgcctct tctacatctg gcgcccctag ccacgacgtg
     1561 gcccagaccc tgcagcagtc cgacgtggaa gattacaccc tgttcaacat gggcgtgcgg
     1621 cggaaggccg catttctgct gcagtcccca accgtgcagg ccacccacca ctggtcccgg
     1681 ctggtcaaga gagccttcgc cggccagagc gagtacgtgc tgcggatcgt gttcgcctac
     1741 ccccacaacg gcagcgtgat gcgggacatc gtgaacgagg aagtgaccgt cggcggcagc
     1801 caggccgact acctgcactt cttcaacaag gcccagccat tcttccactg cgtgaagaaa
     1861 gccgtgcgcg tgatggactg gctgcagaga tacggcgact atggcgcgcc cggcagcctg
     1921 caccacatcc tggacgccca gaagatgctg tggaaccaca gggacaggaa cctgcctcct
     1981 ctggcccctc tgggccccca ccaccatcac caccattgag tctagacacg tgattaatta
     2041 aggatccccc gacctcgacc tctggctaat aaaggaaatt tattttcatt gcaatagtgt
     2101 gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttggtcg
     2161 agatccctcg gagatctcta gctagaggat cgatccccgc cccggacgaa ctaaacctga
     2221 ctacgacatc tctgcccctt cttcgcgggg cagtgcatgt aatcccttca gttggttggt
     2281 acaacttgcc aactgggccc tgttccacat gtgacacggg gggggaccaa acacaaaggg
     2341 gttctctgac tgtagttgac atccttataa atggatgtgc acatttgcca acactgagtg
     2401 gctttcatcc tggagcagac tttgcagtct gtggactgca acacaacatt gcctttatgt
     2461 gtaactcttg gctgaagctc ttacaccaat gctgggggac atgtacctcc caggggccca
     2521 ggaagactac gggaggctac accaacgtca atcagagggg cctgtgtagc taccgataag
     2581 cggaccctca agagggcatt agcaatagtg tttataaggc ccccttgtta accctaaacg
     2641 ggtagcatat gcttcccggg tagtagtata tactatccag actaacccta attcaatagc
     2701 atatgttacc caacgggaag catatgctat cgaattaggg ttagtaaaag ggtcctaagg
     2761 aacagcgata tctcccaccc catgagctgt cacggtttta tttacatggg gtcaggattc
     2821 cacgagggta gtgaaccatt ttagtcacaa gggcagtggc tgaagatcaa ggagcgggca
     2881 gtgaactctc ctgaatcttc gcctgcttct tcattctcct tcgtttagct aatagaataa
     2941 ctgctgagtt gtgaacagta aggtgtatgt gaggtgctcg aaaacaaggt ttcaggtgac
     3001 gcccccagaa taaaatttgg acggggggtt cagtggtggc attgtgctat gacaccaata
     3061 taaccctcac aaaccccttg ggcaataaat actagtgtag gaatgaaaca ttctgaatat
     3121 ctttaacaat agaaatccat ggggtgggga caagccgtaa agactggatg tccatctcac
     3181 acgaatttat ggctatgggc aacacataat cctagtgcaa tatgatactg gggttattaa
     3241 gatgtgtccc aggcagggac caagacaggt gaaccatgtt gttacactct atttgtaaca
     3301 aggggaaaga gagtggacgc cgacagcagc ggactccact ggttgtctct aacacccccg
     3361 aaaattaaac ggggctccac gccaatgggg cccataaaca aagacaagtg gccactcttt
     3421 tttttgaaat tgtggagtgg gggcacgcgt cagcccccac acgccgccct gcggttttgg
     3481 actgtaaaat aagggtgtaa taacttggct gattgtaacc ccgctaacca ctgcggtcaa
     3541 accacttgcc cacaaaacca ctaatggcac cccggggaat acctgcataa gtaggtgggc
     3601 gggccaagat aggggcgcga ttgctgcgat ctggaggaca aattacacac acttgcgcct
     3661 gagcgccaag cacagggttg ttggtcctca tattcacgag gtcgctgaga gcacggtggg
     3721 ctaatgttgc catgggtagc atatactacc caaatatctg gatagcatat gctatcctaa
     3781 tctatatctg ggtagcatag gctatcctaa tctatatctg ggtagcatat gctatcctaa
     3841 tctatatctg ggtagtatat gctatcctaa tttatatctg ggtagcatag gctatcctaa
     3901 tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagtatat gctatcctaa
     3961 tctgtatccg ggtagcatat gctatcctaa tagagattag ggtagtatat gctatcctaa
     4021 tttatatctg ggtagcatat actacccaaa tatctggata gcatatgcta tcctaatcta
     4081 tatctgggta gcatatgcta tcctaatcta tatctgggta gcataggcta tcctaatcta
     4141 tatctgggta gcatatgcta tcctaatcta tatctgggta gtatatgcta tcctaattta
     4201 tatctgggta gcataggcta tcctaatcta tatctgggta gcatatgcta tcctaatcta
     4261 tatctgggta gtatatgcta tcctaatctg tatccgggta gcatatgcta tcctcatgat
     4321 aagctgtcaa acatgagaat tttcttgaag acgaaagggc ctcgtgatac gcctattttt
     4381 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     4441 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     4501 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     4561 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     4621 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     4681 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     4741 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc
     4801 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     4861 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     4921 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     4981 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     5041 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat
     5101 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     5161 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     5221 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     5281 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     5341 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     5401 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     5461 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     5521 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5581 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5641 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     5701 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     5761 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5821 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5881 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5941 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     6001 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     6061 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     6121 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     6181 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     6241 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     6301 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     6361 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     6421 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     6481 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     6541 ataacaattt cacacaggaa acagctatga ccatgattac gccaagctct agctagaggt
     6601 cgaccaattc tcatgtttga cagcttatca tcgcagatcc gggcaacgtt gttgccattg
     6661 ctgcaggcgc agaactggta ggtatggaag atctatacat tgaatcaata ttggcaatta
     6721 gccatattag tcattggtta tatagcataa atcaatattg gctattggcc attgcatacg
     6781 ttgtatctat atcataatat gtacatttat attggctcat gtccaatatg accgccatgt
     6841 tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
     6901 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     6961 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     7021 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     7081 caagtgtatc atatgccaag tccgccccct attgacgtca atgacggtaa atggcccgcc
     7141 tggcattatg cccagtacat gaccttacgg gactttccta cttggcagta catctacgta
     7201 ttagtcatcg ctattaccat g
//
