LOCUS       sequence-274751-        9492 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 9492)
  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 9492)
  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: D49_Ld_320280
FEATURES             Location/Qualifiers
     source          1..9492
                     /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             4270..4287
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4339..4394
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4393..4412)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6630..6648)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6716..6820
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6821..6889,6890..7681)
                     /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(7039..7058)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7852..8440
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8341..8360
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8594..8611
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8728..8749
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8764..8769,8770..8787,8788..8794)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8802..8818
                     /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     8807..8829
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8826..8842
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8826..8842
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        9113..9492
                     /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 gccaccacca agatcaccta cgtgggcgtg
      841 ttccaggcca ccgagggcga gacaaccacc gtgctgctga aagtgggcgc cctgaaggcc
      901 gagtgtatcg ccggcaccct gacctacaac gtggacgacg accacggcaa caaggccacc
      961 ttccccaaca tgcacgaccc ccggatggaa ttcaccgccg ccatcaccga catgaccagc
     1021 ggcgtgcgga ccacctatgt ggacctggtg gtcgtgtgca acgacaccac ccctgtgggc
     1081 ggcaccagcc gggtgtactt ccggatcagc ccccggacca ctaccaccac cctggccccc
     1141 agcatgacct gcaagtctcc aggcgacgcc gtcgtcgtgg gccccatgtc tgccgtgttc
     1201 cccgccaaca cctacaccag cagctgccac catcccagcc tggatatcgc cgacgtgacc
     1261 ggccccagct acgacgccaa gagcctgttc agcgtggaaa tgaccaacaa gagccaggcc
     1321 ctggtgttca acgccaacaa aaccgacacc ctgcccggca agtacaccgc cacaatcctg
     1381 gtgtcctgcg gcagcagctc ctgcgtggtc aagcaggtcg tggtggtgct gccccccaag
     1441 accaccacaa cagccgcccc tgtgatgcag tgccctggca actacaacta cagcctgatc
     1501 ctgaacagcg acggcatcct gatcgagcag atcttcaacc tgagcgccag agccgccgct
     1561 accggcatgt tctgcacctc cggcatcatc agcgccaccc tgaaagaacc cgcccacggg
     1621 gccctgcggt ggaacaacac cggcatcttc gtgtacaccc ctagcaccct ggaagagggc
     1681 gacgatatcg actacttcga gttccagctg acatgcagca acgagggcac caagtgcgcc
     1741 ggaaccgtga tcatctccat cgtgtccgag gaaaaggacg agatctacta cgccatggaa
     1801 ggcgccattg tgtgccgggg cacatgcgac gctgccgcct ggcggacaag ccctaccgcc
     1861 ctggacctgt tcgacgtgaa caagacaggc gtggccgtga cccccagaaa ggacgtgcga
     1921 gccgtggacg gcgtggactt cggctttacc gcctccggcg acctggtcat ccacgcctac
     1981 acaaccatcg gcaacctggc cgctagattc gtgaccttct accctatcac cgaggccagc
     2041 gtgcccggcc tgtttaacag caccgaggtg cccgacggca gccacgtgtc cttcgagccc
     2101 agctgcctca acaagcaggc catcacaggc cagggcaccg atatctggtc ctggaccgac
     2161 ctggacgtgc tgaagggcca cctgaacacc caggccgatg cctacaagag cggcgacaac
     2221 tactaccaga agttcggcgg caaccacaga cactgcgacg tgtacagagt ggacccctgc
     2281 aaatacgccc ccctgatgac ccctaccctg aacgacaaca acaactccgt gtacaacaac
     2341 gccacaccct ccgtgcagtg gaagctgtac atcaacgatt gcgacgccac ctgggtcgga
     2401 aaggcctccg tggaaagcct gcgggccctg agacagccag acggctcccc caccttcaga
     2461 ctggaaggca acggcaccta cctcgtgggg gccgtgtact cccagtccgt gaagcctgcc
     2521 agctggacca gccctctggc cggcatcgtg cacagcgaaa ccgcctacga agtgcggctg
     2581 aagatccaca acatgatcat cgtggacgcc ctggccagac ccagctccga tgccacagcc
     2641 cctctgctgc agatgagcag cgacatccag tacagcgccg gcaagcggac cgatacccag
     2701 gaacggctgt acagatacag cgtcctgctg tacccctact ttgagacaaa caagaccgcc
     2761 gctgacatca cctccctgag cctgaaggtg gtgtccacca cactgctgaa ctccacctgg
     2821 actagcccca gcaaggtgga atgccctatg tgcaccggca ccatgatcag ctgcaccggg
     2881 accggcaatg gcttcgagac agactgcggc gataaggctc tgatgacatt cgaggctctg
     2941 gactacaacg cctccgactt cgagggcagc atcgtgaacg atggctccag cggcaagagc
     3001 gcctccacca gcgtgtcccc aaagaacgct ctgagagtga cctttgccgt gcgcgtgaac
     3061 ggcgctggca gcgctgacgc cacagatgcc tatcccgtgg gcaccttcgc catcagcgtg
     3121 aagctgtcct ccggccaggt gttcgatgtg gtggtcaacc agaccgacta catcagcgag
     3181 ctgaataccc ggtttctgga cacccagatc tgccggccca gcgcctactg gcctgtgcca
     3241 gatcctctgg gcagctctgt gcccgtgaaa ccctactccg ccgacacact ggccttccct
     3301 agcggcagct acaccacaga ggaacaggaa gtgaagcgcg agagcgagtc ctactacggc
     3361 gtgccagagc ggctgacctc ccctagcgtg tgcgtgacac cccccaacgc cgaccggacc
     3421 cacatcgatc tggacaatat gaccgccact atcgagaacg agaagggaca ggtcacagtg
     3481 tccatgtgcg ccgtggccga cgagcggacc ttcggcacca ccgactgggt catgttcagc
     3541 ttccccagaa tccacgagga aacaaaagag atcaacgacg acgagattca gagcatcatc
     3601 gatgacgaaa ccctgagcaa caattcctac accgtggcac agctgcagta cctgaccctg
     3661 tccgtgcacg ccagcgaagt gatgtccagc ctgctggctg gcgacagcgc caacggctat
     3721 gtgaacatcc tgctggacgg ggacaacgcc cccaagtccc agcggggcga ggaagagaca
     3781 tggatgcacc ccgagtcctc cagcagcagc ggccagtctg acgcctacct gccttgggag
     3841 atgtacgcca gcagcctgaa ctaccggcgg atcgcccacc ataccaacgc taacaagtcc
     3901 gaaaccgagc ccttcaactt cgccttcatc cccggctccc tgctgcactc cggaagcagc
     3961 atcagagtgc ccatgattat cagggccgcc attaagtttg tgacctacgc cttcacagcc
     4021 gctaatagca cccacagcag ccagagagtg aagaagtccg agagagagga aaccctcctg
     4081 tatatcgtgc ctgtgtccag gggcatctcc agcgcactga gattcgatac cgacatctac
     4141 accccagtga tcacacagag cggaatcgcc cctggcgcgc ccggcagcct gcaccacatc
     4201 ctggacgccc agaagatgct gtggaaccac agggacagga acctgcctcc tctggcccct
     4261 ctgggccccc accaccatca ccaccattga gtctagacac gtgattaatt aaggatcccc
     4321 cgacctcgac ctctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt
     4381 ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttggtc gagatccctc
     4441 ggagatctct agctagagga tcgatccccg ccccggacga actaaacctg actacgacat
     4501 ctctgcccct tcttcgcggg gcagtgcatg taatcccttc agttggttgg tacaacttgc
     4561 caactgggcc ctgttccaca tgtgacacgg ggggggacca aacacaaagg ggttctctga
     4621 ctgtagttga catccttata aatggatgtg cacatttgcc aacactgagt ggctttcatc
     4681 ctggagcaga ctttgcagtc tgtggactgc aacacaacat tgcctttatg tgtaactctt
     4741 ggctgaagct cttacaccaa tgctggggga catgtacctc ccaggggccc aggaagacta
     4801 cgggaggcta caccaacgtc aatcagaggg gcctgtgtag ctaccgataa gcggaccctc
     4861 aagagggcat tagcaatagt gtttataagg cccccttgtt aaccctaaac gggtagcata
     4921 tgcttcccgg gtagtagtat atactatcca gactaaccct aattcaatag catatgttac
     4981 ccaacgggaa gcatatgcta tcgaattagg gttagtaaaa gggtcctaag gaacagcgat
     5041 atctcccacc ccatgagctg tcacggtttt atttacatgg ggtcaggatt ccacgagggt
     5101 agtgaaccat tttagtcaca agggcagtgg ctgaagatca aggagcgggc agtgaactct
     5161 cctgaatctt cgcctgcttc ttcattctcc ttcgtttagc taatagaata actgctgagt
     5221 tgtgaacagt aaggtgtatg tgaggtgctc gaaaacaagg tttcaggtga cgcccccaga
     5281 ataaaatttg gacggggggt tcagtggtgg cattgtgcta tgacaccaat ataaccctca
     5341 caaacccctt gggcaataaa tactagtgta ggaatgaaac attctgaata tctttaacaa
     5401 tagaaatcca tggggtgggg acaagccgta aagactggat gtccatctca cacgaattta
     5461 tggctatggg caacacataa tcctagtgca atatgatact ggggttatta agatgtgtcc
     5521 caggcaggga ccaagacagg tgaaccatgt tgttacactc tatttgtaac aaggggaaag
     5581 agagtggacg ccgacagcag cggactccac tggttgtctc taacaccccc gaaaattaaa
     5641 cggggctcca cgccaatggg gcccataaac aaagacaagt ggccactctt ttttttgaaa
     5701 ttgtggagtg ggggcacgcg tcagccccca cacgccgccc tgcggttttg gactgtaaaa
     5761 taagggtgta ataacttggc tgattgtaac cccgctaacc actgcggtca aaccacttgc
     5821 ccacaaaacc actaatggca ccccggggaa tacctgcata agtaggtggg cgggccaaga
     5881 taggggcgcg attgctgcga tctggaggac aaattacaca cacttgcgcc tgagcgccaa
     5941 gcacagggtt gttggtcctc atattcacga ggtcgctgag agcacggtgg gctaatgttg
     6001 ccatgggtag catatactac ccaaatatct ggatagcata tgctatccta atctatatct
     6061 gggtagcata ggctatccta atctatatct gggtagcata tgctatccta atctatatct
     6121 gggtagtata tgctatccta atttatatct gggtagcata ggctatccta atctatatct
     6181 gggtagcata tgctatccta atctatatct gggtagtata tgctatccta atctgtatcc
     6241 gggtagcata tgctatccta atagagatta gggtagtata tgctatccta atttatatct
     6301 gggtagcata tactacccaa atatctggat agcatatgct atcctaatct atatctgggt
     6361 agcatatgct atcctaatct atatctgggt agcataggct atcctaatct atatctgggt
     6421 agcatatgct atcctaatct atatctgggt agtatatgct atcctaattt atatctgggt
     6481 agcataggct atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt
     6541 agtatatgct atcctaatct gtatccgggt agcatatgct atcctcatga taagctgtca
     6601 aacatgagaa ttttcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     6661 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     6721 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     6781 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     6841 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     6901 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     6961 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     7021 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga
     7081 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     7141 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     7201 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     7261 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     7321 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac
     7381 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     7441 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     7501 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     7561 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     7621 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     7681 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     7741 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     7801 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     7861 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     7921 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     7981 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     8041 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     8101 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     8161 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     8221 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     8281 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     8341 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     8401 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     8461 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     8521 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     8581 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     8641 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     8701 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
     8761 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     8821 tcacacagga aacagctatg accatgatta cgccaagctc tagctagagg tcgaccaatt
     8881 ctcatgtttg acagcttatc atcgcagatc cgggcaacgt tgttgccatt gctgcaggcg
     8941 cagaactggt aggtatggaa gatctataca ttgaatcaat attggcaatt agccatatta
     9001 gtcattggtt atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta
     9061 tatcataata tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga
     9121 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     9181 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     9241 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     9301 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     9361 catatgccaa gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     9421 gcccagtaca tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc
     9481 gctattacca tg
//
