LOCUS       sequence-274533-        6852 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 6852)
  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 6852)
  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: D20_Ld_140560
FEATURES             Location/Qualifiers
     source          1..6852
                     /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             1630..1647
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1699..1754
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1753..1772)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3990..4008)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4076..4180
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4181..4249,4250..5041)
                     /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(4399..4418)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5212..5800
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5701..5720
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5954..5971
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6088..6109
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6124..6129,6130..6147,6148..6154)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6162..6178
                     /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     6167..6189
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6186..6202
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6186..6202
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        6473..6852
                     /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 gcccttctct gatgctgcct
      841 ggcgtgctgc acgccgctgc cctgctgccc agcagagatc tgtttctggg cagagccggc
      901 agcacctaca actacaccat gtacagcggc agcgccgtga ccggcgagag ccactttacc
      961 ggcctgacca cctaccccta cgtgttcagc gccgacatga acagcgccat cgtggccatg
     1021 ctgaacggct ttctgagcag cgccaagagc aacgtgtccg ccgtgggcgg aggcttccac
     1081 gagtgcatcg acaacaagct gggcgccgtg gacggcacca cctgtttcaa gtctctgctg
     1141 gctgccgccc ctcccgccga ttgtcctgcc gccaacaaac tgtgctgccc cctgttttgg
     1201 agagtgggca gccagggcct gcggaccacc atggctcagg tcatccccca gaccctgttc
     1261 tggaacggcg ccacctccgt gaatggcttc tacaccaact tcagcagaag ctacctggac
     1321 gccaaccgga actgcagcaa gcccctgctg agaaccacca gcctgatcaa cctggccatg
     1381 gacgcctacg gcgagtggcg cttcgtggac ctggtcacca catacaccct gctggaaacc
     1441 gccatcatca acagaaacgc cagctccgcc acccccatct tcggcgagaa cccccaggac
     1501 gccatcacca gcaccaccag atctcagcag cctggcgcgc ccggcagcct gcaccacatc
     1561 ctggacgccc agaagatgct gtggaaccac agggacagga acctgcctcc tctggcccct
     1621 ctgggccccc accaccatca ccaccattga gtctagacac gtgattaatt aaggatcccc
     1681 cgacctcgac ctctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt
     1741 ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttggtc gagatccctc
     1801 ggagatctct agctagagga tcgatccccg ccccggacga actaaacctg actacgacat
     1861 ctctgcccct tcttcgcggg gcagtgcatg taatcccttc agttggttgg tacaacttgc
     1921 caactgggcc ctgttccaca tgtgacacgg ggggggacca aacacaaagg ggttctctga
     1981 ctgtagttga catccttata aatggatgtg cacatttgcc aacactgagt ggctttcatc
     2041 ctggagcaga ctttgcagtc tgtggactgc aacacaacat tgcctttatg tgtaactctt
     2101 ggctgaagct cttacaccaa tgctggggga catgtacctc ccaggggccc aggaagacta
     2161 cgggaggcta caccaacgtc aatcagaggg gcctgtgtag ctaccgataa gcggaccctc
     2221 aagagggcat tagcaatagt gtttataagg cccccttgtt aaccctaaac gggtagcata
     2281 tgcttcccgg gtagtagtat atactatcca gactaaccct aattcaatag catatgttac
     2341 ccaacgggaa gcatatgcta tcgaattagg gttagtaaaa gggtcctaag gaacagcgat
     2401 atctcccacc ccatgagctg tcacggtttt atttacatgg ggtcaggatt ccacgagggt
     2461 agtgaaccat tttagtcaca agggcagtgg ctgaagatca aggagcgggc agtgaactct
     2521 cctgaatctt cgcctgcttc ttcattctcc ttcgtttagc taatagaata actgctgagt
     2581 tgtgaacagt aaggtgtatg tgaggtgctc gaaaacaagg tttcaggtga cgcccccaga
     2641 ataaaatttg gacggggggt tcagtggtgg cattgtgcta tgacaccaat ataaccctca
     2701 caaacccctt gggcaataaa tactagtgta ggaatgaaac attctgaata tctttaacaa
     2761 tagaaatcca tggggtgggg acaagccgta aagactggat gtccatctca cacgaattta
     2821 tggctatggg caacacataa tcctagtgca atatgatact ggggttatta agatgtgtcc
     2881 caggcaggga ccaagacagg tgaaccatgt tgttacactc tatttgtaac aaggggaaag
     2941 agagtggacg ccgacagcag cggactccac tggttgtctc taacaccccc gaaaattaaa
     3001 cggggctcca cgccaatggg gcccataaac aaagacaagt ggccactctt ttttttgaaa
     3061 ttgtggagtg ggggcacgcg tcagccccca cacgccgccc tgcggttttg gactgtaaaa
     3121 taagggtgta ataacttggc tgattgtaac cccgctaacc actgcggtca aaccacttgc
     3181 ccacaaaacc actaatggca ccccggggaa tacctgcata agtaggtggg cgggccaaga
     3241 taggggcgcg attgctgcga tctggaggac aaattacaca cacttgcgcc tgagcgccaa
     3301 gcacagggtt gttggtcctc atattcacga ggtcgctgag agcacggtgg gctaatgttg
     3361 ccatgggtag catatactac ccaaatatct ggatagcata tgctatccta atctatatct
     3421 gggtagcata ggctatccta atctatatct gggtagcata tgctatccta atctatatct
     3481 gggtagtata tgctatccta atttatatct gggtagcata ggctatccta atctatatct
     3541 gggtagcata tgctatccta atctatatct gggtagtata tgctatccta atctgtatcc
     3601 gggtagcata tgctatccta atagagatta gggtagtata tgctatccta atttatatct
     3661 gggtagcata tactacccaa atatctggat agcatatgct atcctaatct atatctgggt
     3721 agcatatgct atcctaatct atatctgggt agcataggct atcctaatct atatctgggt
     3781 agcatatgct atcctaatct atatctgggt agtatatgct atcctaattt atatctgggt
     3841 agcataggct atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt
     3901 agtatatgct atcctaatct gtatccgggt agcatatgct atcctcatga taagctgtca
     3961 aacatgagaa ttttcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     4021 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     4081 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4141 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     4201 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     4261 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     4321 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     4381 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga
     4441 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     4501 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     4561 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     4621 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     4681 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac
     4741 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     4801 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     4861 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     4921 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     4981 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     5041 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     5101 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     5161 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     5221 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     5281 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     5341 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     5401 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     5461 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5521 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     5581 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     5641 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     5701 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     5761 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     5821 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     5881 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     5941 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     6001 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     6061 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
     6121 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     6181 tcacacagga aacagctatg accatgatta cgccaagctc tagctagagg tcgaccaatt
     6241 ctcatgtttg acagcttatc atcgcagatc cgggcaacgt tgttgccatt gctgcaggcg
     6301 cagaactggt aggtatggaa gatctataca ttgaatcaat attggcaatt agccatatta
     6361 gtcattggtt atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta
     6421 tatcataata tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga
     6481 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     6541 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     6601 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     6661 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     6721 catatgccaa gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     6781 gcccagtaca tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc
     6841 gctattacca tg
//
