LOCUS       sequence-274874-        7956 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 7956)
  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 7956)
  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: D68_Ld_161580
FEATURES             Location/Qualifiers
     source          1..7956
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     1086..1105
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1128..1137
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3114..3131
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3183..3238
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3237..3256)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5474..5492)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5560..5664
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5665..5733,5734..6525)
                     /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(5883..5902)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6696..7284
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7185..7204
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7438..7455
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7572..7593
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7608..7613,7614..7631,7632..7638)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7646..7662
                     /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     7651..7673
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7670..7686
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7670..7686
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agatcctcac tctcttccgc atcgctgtct gcgagggcca gctgttgggc tcgcggttga
      661 ggacaaactc ttcgcggtct ttccagtact cttggatcgg aaacccgtcg gcctccgaac
      721 ggtactccgc caccgaggga cctgagcgag tccgcatcga ccggatcgga aaacctctcg
      781 agaaaggcgt ctaaccagtc acagtcgcaa ggtaggctga gcaccgtggc gggcggcagc
      841 gggtggcggt cggggttgtt tctggcggag gtgctgctga tgatgtaatt aaagtaggcg
      901 gtcttgagac ggcggatggt cgaggtgagg tgtggcaggc ttgagatcca gctgttgggg
      961 tgagtactcc ctctcaaaag cgggcattac ttctgcgcta agattgtcag tttccaaaaa
     1021 cgaggaggat ttgatattca cctggcccga tctggccata cacttgagtg acaatgacat
     1081 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caagtttgcc accatggagt
     1141 ttcagaccca ggtactcatg tccctgctgc tctgcatgtc tggtgcggcc gccctgtctc
     1201 tcacctctgg ctccggaact atgggcgtgt gttgctccga ctgcaagaga agtgatgctg
     1261 aaaagaacac caaaagcgtt gggcaggtca ccacgtcatc cggctttaag cctatgcaaa
     1321 attctgccct ttgtttccag cccgtggcgt tcgacgtcgt agtgccagag ggtgctaata
     1381 tgatgacagc atctccacag agagggtccg tggtggtgga ctgtgaagtc aacaatttct
     1441 caggcaccct ttccgatgcc tctctgcata cgattcggga cacgattaca gttgatcgca
     1501 gtgggaacac ctacgaccca atgaacgaaa cacttagcaa catgtccttc acaaacagtt
     1561 ttcacgctgt cgacaagcga gcagtgcaga ttttcatgaa gaacgtcgcc agactcgccg
     1621 agcggaataa taaaggtacg atgctggaag agtctctgat cagcgcttcc ataggtcctg
     1681 atacctgcgc ctgcaacaga agtatgctgg gatccctgaa catgtccaat atggcccaag
     1741 aggatctcga ctatatcgct gccggggcta cactccagga caggattgct cggctcgagg
     1801 aagttgaggc ggtgctcagg cggtccatcg acgggtcctg gagagtggac tcaatgcttc
     1861 tccaactggc ttgctgccaa atacttccaa cactgagaca gcgaaggtgt ccctgggctg
     1921 ccggacggtc agcaacccaa tcaaagtcaa ccgctgcgca gcagcagtcc aaaagaaagc
     1981 ctgtgctcgt catctacgct cagggccttc aaatggatgc ttctgtgatt accgcaccca
     2041 cttccattac cgccagtcgc gcaagcccta ccttgccgtc tgaactgtct atcggcaacc
     2101 tcaacgatgg gaaggaggga ggcggcaata atacgagtat ggtgctgggc gcctctcgga
     2161 gcagagccag cgtttctact gccgccacca aggtgcagca gtacgaaatg ctggccctta
     2221 cctgtcgcga catcgatccc catcgtgccg acagcgcgtt ggtggagaac aacacctggg
     2281 actttctgtc tgttagcagg gccgccgcaa ctctgaacga gggcaggtgt atcttccgtt
     2341 tcctttgcgg tagcctggca gaggctgatc cagagactca gggtattgtg gatgctatca
     2401 ttaatcttag tagcttcgcc gatgaagtga aggagaataa tgtgccaaag gtgagcgtgt
     2461 acaacagtga ggatgccgcc aaatacatcg gcatccacgt gaaggctatg tacatgctcc
     2521 ccggaattat gctcaaggat ataacgaaag atttggtcgt gtccgccgag tatcagaagg
     2581 cgtgccagcc cagcgagaac aaaatccagg aatgtgaact tgacgacacc ttccgcctga
     2641 ccatgaccat tattcgcttt gtctttagcc tcacaatcca attcaaagtc tcagagatca
     2701 ctgatcccgc cgtattgcgc tccctggccg tatgcgatgg aatggtaccc gccaaagccc
     2761 tgatgctgga gagaacaaag cgacatatcg ccaaagttga cgctaccgcc aaagtcagat
     2821 caatgctgct ctattatccc gtgaacgatg gtattcttgt ggacaatcat acggtggtgc
     2881 tgaacaccag cctgccgaaa gtagtcagca agattatgaa taattttggt agtcaagggg
     2941 cgacacagag tgtccaaact gccaagctga cccgggagta tctgctccag aggttcggtg
     3001 acagcagatc caaatacggc gcgcccggca gcctgcacca catcctggac gcccagaaga
     3061 tgctgtggaa ccacagggac aggaacctgc ctcctctggc ccctctgggc ccccaccacc
     3121 atcaccacca ttgagtctag acacgtgatt aattaaggat cccccgacct cgacctctgg
     3181 ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact
     3241 cggaaggaca tatgggaggg caaatcattt ggtcgagatc cctcggagat ctctagctag
     3301 aggatcgatc cccgccccgg acgaactaaa cctgactacg acatctctgc cccttcttcg
     3361 cggggcagtg catgtaatcc cttcagttgg ttggtacaac ttgccaactg ggccctgttc
     3421 cacatgtgac acgggggggg accaaacaca aaggggttct ctgactgtag ttgacatcct
     3481 tataaatgga tgtgcacatt tgccaacact gagtggcttt catcctggag cagactttgc
     3541 agtctgtgga ctgcaacaca acattgcctt tatgtgtaac tcttggctga agctcttaca
     3601 ccaatgctgg gggacatgta cctcccaggg gcccaggaag actacgggag gctacaccaa
     3661 cgtcaatcag aggggcctgt gtagctaccg ataagcggac cctcaagagg gcattagcaa
     3721 tagtgtttat aaggccccct tgttaaccct aaacgggtag catatgcttc ccgggtagta
     3781 gtatatacta tccagactaa ccctaattca atagcatatg ttacccaacg ggaagcatat
     3841 gctatcgaat tagggttagt aaaagggtcc taaggaacag cgatatctcc caccccatga
     3901 gctgtcacgg ttttatttac atggggtcag gattccacga gggtagtgaa ccattttagt
     3961 cacaagggca gtggctgaag atcaaggagc gggcagtgaa ctctcctgaa tcttcgcctg
     4021 cttcttcatt ctccttcgtt tagctaatag aataactgct gagttgtgaa cagtaaggtg
     4081 tatgtgaggt gctcgaaaac aaggtttcag gtgacgcccc cagaataaaa tttggacggg
     4141 gggttcagtg gtggcattgt gctatgacac caatataacc ctcacaaacc ccttgggcaa
     4201 taaatactag tgtaggaatg aaacattctg aatatcttta acaatagaaa tccatggggt
     4261 ggggacaagc cgtaaagact ggatgtccat ctcacacgaa tttatggcta tgggcaacac
     4321 ataatcctag tgcaatatga tactggggtt attaagatgt gtcccaggca gggaccaaga
     4381 caggtgaacc atgttgttac actctatttg taacaagggg aaagagagtg gacgccgaca
     4441 gcagcggact ccactggttg tctctaacac ccccgaaaat taaacggggc tccacgccaa
     4501 tggggcccat aaacaaagac aagtggccac tctttttttt gaaattgtgg agtgggggca
     4561 cgcgtcagcc cccacacgcc gccctgcggt tttggactgt aaaataaggg tgtaataact
     4621 tggctgattg taaccccgct aaccactgcg gtcaaaccac ttgcccacaa aaccactaat
     4681 ggcaccccgg ggaatacctg cataagtagg tgggcgggcc aagatagggg cgcgattgct
     4741 gcgatctgga ggacaaatta cacacacttg cgcctgagcg ccaagcacag ggttgttggt
     4801 cctcatattc acgaggtcgc tgagagcacg gtgggctaat gttgccatgg gtagcatata
     4861 ctacccaaat atctggatag catatgctat cctaatctat atctgggtag cataggctat
     4921 cctaatctat atctgggtag catatgctat cctaatctat atctgggtag tatatgctat
     4981 cctaatttat atctgggtag cataggctat cctaatctat atctgggtag catatgctat
     5041 cctaatctat atctgggtag tatatgctat cctaatctgt atccgggtag catatgctat
     5101 cctaatagag attagggtag tatatgctat cctaatttat atctgggtag catatactac
     5161 ccaaatatct ggatagcata tgctatccta atctatatct gggtagcata tgctatccta
     5221 atctatatct gggtagcata ggctatccta atctatatct gggtagcata tgctatccta
     5281 atctatatct gggtagtata tgctatccta atttatatct gggtagcata ggctatccta
     5341 atctatatct gggtagcata tgctatccta atctatatct gggtagtata tgctatccta
     5401 atctgtatcc gggtagcata tgctatcctc atgataagct gtcaaacatg agaattttct
     5461 tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     5521 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     5581 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     5641 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     5701 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     5761 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     5821 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     5881 ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc
     5941 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     6001 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     6061 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     6121 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     6181 aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta
     6241 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     6301 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     6361 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     6421 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     6481 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     6541 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     6601 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     6661 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6721 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6781 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6841 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6901 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6961 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     7021 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     7081 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     7141 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     7201 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     7261 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     7321 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     7381 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     7441 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     7501 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     7561 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     7621 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     7681 tatgaccatg attacgccaa gctctagcta gaggtcgacc aattctcatg tttgacagct
     7741 tatcatcgca gatccgggca acgttgttgc cattgctgca ggcgcagaac tggtaggtat
     7801 ggaagatcta tacattgaat caatattggc aattagccat attagtcatt ggttatatag
     7861 cataaatcaa tattggctat tggccattgc atacgttgta tctatatcat aatatgtaca
     7921 tttatattgg ctcatgtcca atatgaccgc catgtt
//
