LOCUS       sequence-274550-        7632 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 7632)
  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 7632)
  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: D31_Ld_261020
FEATURES             Location/Qualifiers
     source          1..7632
                     /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             2410..2427
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2479..2534
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2533..2552)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4770..4788)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4856..4960
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4961..5029,5030..5821)
                     /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(5179..5198)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5992..6580
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6481..6500
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6734..6751
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6868..6889
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6904..6909,6910..6927,6928..6934)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6942..6958
                     /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     6947..6969
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6966..6982
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6966..6982
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7253..7632
                     /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 gccaccggac ctgtgctgga agtgggagtg
      841 gaccgggcca tcaccttcaa cagacagctg ggcgacggcg acgagctgct gttcggaagc
      901 agagccgtgc tgctgcgggg cgagatcgtg cggtatacag ccggcgcttc cgccagctac
      961 cacggctaca ccgtgtacgt gcagtgcagc gagcacctga ccaacttcgc cctgcagcag
     1021 ctggccgctc ccgagctgtc taaggccgcc aagggcctgg tggtggaact gtgcgacacc
     1081 gacaccacca acgaggacgt gctgagcttc ttcttctcca acgccgctgc cagcatcccc
     1141 gtgtactttc tgcccaacgg caccgccagc aaagagctga gcagactgct gagcgtggcc
     1201 gcccagagaa gcgccaccga gagggtggtg ctgtccgtgg ccaagaccag gcggctgagc
     1261 gagctggtgg aaaacagcac cctgccctcc gccaccatcg agagccacta cgtgcacaag
     1321 cccaagcaga ccagaagcaa agccgccagc gctgccgccg ctcctcaggt gctggtcacc
     1381 gcccacttcg actccctggg cgtgtcccca gccagcagaa cctctggcgg aaccagcgga
     1441 gccgtggtgg ccatggaact gtggcggaga ctgaccagca ccccctacgc cagacaggaa
     1501 agcgtcgccc cttacgaagt gaccgtcctg ctgggcagca ccagccggtt caactacgcc
     1561 ggcaccacaa gctggctgag ccagcacacc gatagagagc tggaccagtt ccgggtggtc
     1621 ctgtgcctgg acgaactgct gccccccaga gagacaagcg agaacgcccc cgacctgtat
     1681 ctgcatgtgc aggatgtgct gatgaagcgg cagcacggcc agcaggtcgt ggaacaggtg
     1741 gaagccgccg caaagctgct gggaatcagc ctgaaggtgg tgtccgccaa gacaaactac
     1801 cagcactacg acctggaatt cgagcacgag gccttcgcca gccggcagat gaccgccatg
     1861 accctgagca cccaccgggt gcaccataac gaccagctgt tccgggacct gcggaggcct
     1921 cctgccacag ccgctgatgc cgctatgctg gccaagcggg ccgacctggt cgaggccatt
     1981 gtgcggattg tggccgaggc tgcccctagc gccgagggcg tgacaacagt gtggcctggc
     2041 gccgctagct acatccaggg catgctgcac tacgcctccg acagccagag atcccccgtg
     2101 gcccagaacg gcgctggcct gagacagtac gccgccacag tggaacacca catgagagcc
     2161 caggccgctg ctgctcagag agccgccttt accagcgcca gctctgtggc cacctaccag
     2221 gaactgcgga cccccggcat caccctgttc ggcccatacg aagagacaat gcaggtgttc
     2281 atggccaagt cctacctgtt cgagtgcgcc gtgggcgcgc ccggcagcct gcaccacatc
     2341 ctggacgccc agaagatgct gtggaaccac agggacagga acctgcctcc tctggcccct
     2401 ctgggccccc accaccatca ccaccattga gtctagacac gtgattaatt aaggatcccc
     2461 cgacctcgac ctctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt
     2521 ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttggtc gagatccctc
     2581 ggagatctct agctagagga tcgatccccg ccccggacga actaaacctg actacgacat
     2641 ctctgcccct tcttcgcggg gcagtgcatg taatcccttc agttggttgg tacaacttgc
     2701 caactgggcc ctgttccaca tgtgacacgg ggggggacca aacacaaagg ggttctctga
     2761 ctgtagttga catccttata aatggatgtg cacatttgcc aacactgagt ggctttcatc
     2821 ctggagcaga ctttgcagtc tgtggactgc aacacaacat tgcctttatg tgtaactctt
     2881 ggctgaagct cttacaccaa tgctggggga catgtacctc ccaggggccc aggaagacta
     2941 cgggaggcta caccaacgtc aatcagaggg gcctgtgtag ctaccgataa gcggaccctc
     3001 aagagggcat tagcaatagt gtttataagg cccccttgtt aaccctaaac gggtagcata
     3061 tgcttcccgg gtagtagtat atactatcca gactaaccct aattcaatag catatgttac
     3121 ccaacgggaa gcatatgcta tcgaattagg gttagtaaaa gggtcctaag gaacagcgat
     3181 atctcccacc ccatgagctg tcacggtttt atttacatgg ggtcaggatt ccacgagggt
     3241 agtgaaccat tttagtcaca agggcagtgg ctgaagatca aggagcgggc agtgaactct
     3301 cctgaatctt cgcctgcttc ttcattctcc ttcgtttagc taatagaata actgctgagt
     3361 tgtgaacagt aaggtgtatg tgaggtgctc gaaaacaagg tttcaggtga cgcccccaga
     3421 ataaaatttg gacggggggt tcagtggtgg cattgtgcta tgacaccaat ataaccctca
     3481 caaacccctt gggcaataaa tactagtgta ggaatgaaac attctgaata tctttaacaa
     3541 tagaaatcca tggggtgggg acaagccgta aagactggat gtccatctca cacgaattta
     3601 tggctatggg caacacataa tcctagtgca atatgatact ggggttatta agatgtgtcc
     3661 caggcaggga ccaagacagg tgaaccatgt tgttacactc tatttgtaac aaggggaaag
     3721 agagtggacg ccgacagcag cggactccac tggttgtctc taacaccccc gaaaattaaa
     3781 cggggctcca cgccaatggg gcccataaac aaagacaagt ggccactctt ttttttgaaa
     3841 ttgtggagtg ggggcacgcg tcagccccca cacgccgccc tgcggttttg gactgtaaaa
     3901 taagggtgta ataacttggc tgattgtaac cccgctaacc actgcggtca aaccacttgc
     3961 ccacaaaacc actaatggca ccccggggaa tacctgcata agtaggtggg cgggccaaga
     4021 taggggcgcg attgctgcga tctggaggac aaattacaca cacttgcgcc tgagcgccaa
     4081 gcacagggtt gttggtcctc atattcacga ggtcgctgag agcacggtgg gctaatgttg
     4141 ccatgggtag catatactac ccaaatatct ggatagcata tgctatccta atctatatct
     4201 gggtagcata ggctatccta atctatatct gggtagcata tgctatccta atctatatct
     4261 gggtagtata tgctatccta atttatatct gggtagcata ggctatccta atctatatct
     4321 gggtagcata tgctatccta atctatatct gggtagtata tgctatccta atctgtatcc
     4381 gggtagcata tgctatccta atagagatta gggtagtata tgctatccta atttatatct
     4441 gggtagcata tactacccaa atatctggat agcatatgct atcctaatct atatctgggt
     4501 agcatatgct atcctaatct atatctgggt agcataggct atcctaatct atatctgggt
     4561 agcatatgct atcctaatct atatctgggt agtatatgct atcctaattt atatctgggt
     4621 agcataggct atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt
     4681 agtatatgct atcctaatct gtatccgggt agcatatgct atcctcatga taagctgtca
     4741 aacatgagaa ttttcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     4801 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     4861 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4921 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     4981 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     5041 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     5101 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     5161 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga
     5221 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     5281 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     5341 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     5401 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     5461 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac
     5521 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     5581 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     5641 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     5701 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     5761 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     5821 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     5881 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     5941 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     6001 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     6061 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     6121 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     6181 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     6241 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     6301 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     6361 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     6421 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     6481 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     6541 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     6601 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     6661 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     6721 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     6781 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     6841 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
     6901 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     6961 tcacacagga aacagctatg accatgatta cgccaagctc tagctagagg tcgaccaatt
     7021 ctcatgtttg acagcttatc atcgcagatc cgggcaacgt tgttgccatt gctgcaggcg
     7081 cagaactggt aggtatggaa gatctataca ttgaatcaat attggcaatt agccatatta
     7141 gtcattggtt atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta
     7201 tatcataata tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga
     7261 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     7321 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     7381 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     7441 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     7501 catatgccaa gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     7561 gcccagtaca tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc
     7621 gctattacca tg
//
