LOCUS       sequence-274872-        7122 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 7122)
  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 7122)
  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: D69_Ld_191460
FEATURES             Location/Qualifiers
     source          1..7122
                     /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             1900..1917
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1969..2024
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2023..2042)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4260..4278)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4346..4450
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4451..4519,4520..5311)
                     /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(4669..4688)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5482..6070
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5971..5990
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6224..6241
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6358..6379
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6394..6399,6400..6417,6418..6424)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6432..6448
                     /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     6437..6459
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6456..6472
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6456..6472
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        6743..7122
                     /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 gccctgatcg cacagacacc cctgggagtc
      841 gatgatttca ttgcatccgc acactacggt cgattcaaga agcgccacgg gaagcctttc
      901 ggcgaagatg ctgaagaggg cagacgcttc aatgctttta agcaaaatat gcagacagcc
      961 tatttcctga acgcgcataa cccccatgcc cattatgatg tgtcaggcaa gtttgctgat
     1021 ctgactccac aggaattcgc aaaactttac ctcaacccga actactacgc ccggcatgga
     1081 aaggattaca aggaacacgt tcacgtcgac gatagtgttc ggagtggagt gatgagcgtt
     1141 gactggcgcg aaaagggagt ggtcactccc gtgaagaacc agggtatgtg cgggagctgt
     1201 tgggccttcg ccacaaccgg taatatcgag ggccagtggg ccctgaagaa ccactcactg
     1261 gtgagcctct ccgagcaggt gcttgtgagc tgtgacaaca tcgatgacgg gtgtaacggt
     1321 ggactgatgg aacaagcaat gcagtggatc ataaacgatc acaacggcac tgtgcctaca
     1381 gaagatagtt acccctacac atccgcgggc ggtactaggc ccccgtgcca cgataacgga
     1441 actgtgggag ctaagatcgc tggatacatg agtctgcccc atgacgaaga ggaaattgcc
     1501 gcctacgtcg gaaagaacgg acctgtcgcc gtagcagtcg atgccacaac ctggcagctc
     1561 tatttcggcg gcgtggttac cctctgcttc ggactcagcc ttaaccacgg agtgttggtc
     1621 gtcggattta accgtcaggc caagccacca tactggattg ttaagaacag ttggggctct
     1681 tcctggggtg agaaaggcta catccgactg gctatgggct caaaccagtg tcttctgaag
     1741 aattatgcag tcaccgcaac aattgacgat agcaacacct ctcacgtgcc tactacagcc
     1801 gccggcgcgc ccggcagcct gcaccacatc ctggacgccc agaagatgct gtggaaccac
     1861 agggacagga acctgcctcc tctggcccct ctgggccccc accaccatca ccaccattga
     1921 gtctagacac gtgattaatt aaggatcccc cgacctcgac ctctggctaa taaaggaaat
     1981 ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg
     2041 ggagggcaaa tcatttggtc gagatccctc ggagatctct agctagagga tcgatccccg
     2101 ccccggacga actaaacctg actacgacat ctctgcccct tcttcgcggg gcagtgcatg
     2161 taatcccttc agttggttgg tacaacttgc caactgggcc ctgttccaca tgtgacacgg
     2221 ggggggacca aacacaaagg ggttctctga ctgtagttga catccttata aatggatgtg
     2281 cacatttgcc aacactgagt ggctttcatc ctggagcaga ctttgcagtc tgtggactgc
     2341 aacacaacat tgcctttatg tgtaactctt ggctgaagct cttacaccaa tgctggggga
     2401 catgtacctc ccaggggccc aggaagacta cgggaggcta caccaacgtc aatcagaggg
     2461 gcctgtgtag ctaccgataa gcggaccctc aagagggcat tagcaatagt gtttataagg
     2521 cccccttgtt aaccctaaac gggtagcata tgcttcccgg gtagtagtat atactatcca
     2581 gactaaccct aattcaatag catatgttac ccaacgggaa gcatatgcta tcgaattagg
     2641 gttagtaaaa gggtcctaag gaacagcgat atctcccacc ccatgagctg tcacggtttt
     2701 atttacatgg ggtcaggatt ccacgagggt agtgaaccat tttagtcaca agggcagtgg
     2761 ctgaagatca aggagcgggc agtgaactct cctgaatctt cgcctgcttc ttcattctcc
     2821 ttcgtttagc taatagaata actgctgagt tgtgaacagt aaggtgtatg tgaggtgctc
     2881 gaaaacaagg tttcaggtga cgcccccaga ataaaatttg gacggggggt tcagtggtgg
     2941 cattgtgcta tgacaccaat ataaccctca caaacccctt gggcaataaa tactagtgta
     3001 ggaatgaaac attctgaata tctttaacaa tagaaatcca tggggtgggg acaagccgta
     3061 aagactggat gtccatctca cacgaattta tggctatggg caacacataa tcctagtgca
     3121 atatgatact ggggttatta agatgtgtcc caggcaggga ccaagacagg tgaaccatgt
     3181 tgttacactc tatttgtaac aaggggaaag agagtggacg ccgacagcag cggactccac
     3241 tggttgtctc taacaccccc gaaaattaaa cggggctcca cgccaatggg gcccataaac
     3301 aaagacaagt ggccactctt ttttttgaaa ttgtggagtg ggggcacgcg tcagccccca
     3361 cacgccgccc tgcggttttg gactgtaaaa taagggtgta ataacttggc tgattgtaac
     3421 cccgctaacc actgcggtca aaccacttgc ccacaaaacc actaatggca ccccggggaa
     3481 tacctgcata agtaggtggg cgggccaaga taggggcgcg attgctgcga tctggaggac
     3541 aaattacaca cacttgcgcc tgagcgccaa gcacagggtt gttggtcctc atattcacga
     3601 ggtcgctgag agcacggtgg gctaatgttg ccatgggtag catatactac ccaaatatct
     3661 ggatagcata tgctatccta atctatatct gggtagcata ggctatccta atctatatct
     3721 gggtagcata tgctatccta atctatatct gggtagtata tgctatccta atttatatct
     3781 gggtagcata ggctatccta atctatatct gggtagcata tgctatccta atctatatct
     3841 gggtagtata tgctatccta atctgtatcc gggtagcata tgctatccta atagagatta
     3901 gggtagtata tgctatccta atttatatct gggtagcata tactacccaa atatctggat
     3961 agcatatgct atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt
     4021 agcataggct atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt
     4081 agtatatgct atcctaattt atatctgggt agcataggct atcctaatct atatctgggt
     4141 agcatatgct atcctaatct atatctgggt agtatatgct atcctaatct gtatccgggt
     4201 agcatatgct atcctcatga taagctgtca aacatgagaa ttttcttgaa gacgaaaggg
     4261 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     4321 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     4381 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     4441 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     4501 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     4561 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     4621 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     4681 ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     4741 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     4801 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     4861 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     4921 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     4981 caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     5041 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     5101 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     5161 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     5221 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     5281 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     5341 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     5401 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     5461 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     5521 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     5581 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     5641 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     5701 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     5761 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     5821 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     5881 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     5941 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     6001 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     6061 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     6121 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     6181 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     6241 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     6301 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     6361 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     6421 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     6481 cgccaagctc tagctagagg tcgaccaatt ctcatgtttg acagcttatc atcgcagatc
     6541 cgggcaacgt tgttgccatt gctgcaggcg cagaactggt aggtatggaa gatctataca
     6601 ttgaatcaat attggcaatt agccatatta gtcattggtt atatagcata aatcaatatt
     6661 ggctattggc cattgcatac gttgtatcta tatcataata tgtacattta tattggctca
     6721 tgtccaatat gaccgccatg ttgacattga ttattgacta gttattaata gtaatcaatt
     6781 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     6841 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     6901 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     6961 actgcccact tggcagtaca tcaagtgtat catatgccaa gtccgccccc tattgacgtc
     7021 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttacg ggactttcct
     7081 acttggcagt acatctacgt attagtcatc gctattacca tg
//
