LOCUS       sequence-262349-        8616 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 8616)
  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 8616)
  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: D57_Ld_190640
FEATURES             Location/Qualifiers
     source          1..8616
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        7..386
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        387..590
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     541..561
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     587..611
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     1092..1111
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1134..1143
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3780..3797
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3849..3904
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3903..3922)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6140..6158)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6226..6330
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6331..6399,6400..7191)
                     /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(6549..6568)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7362..7950
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7851..7870
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8104..8121
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8238..8259
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8274..8279,8280..8297,8298..8304)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8312..8328
                     /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     8317..8339
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8336..8352
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8336..8352
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 catgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
       61 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      121 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      181 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      241 tacatcaagt gtatcatatg ccaagtccgc cccctattga cgtcaatgac ggtaaatggc
      301 ccgcctggca ttatgcccag tacatgacct tacgggactt tcctacttgg cagtacatct
      361 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacacc aatgggcgtg
      421 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      481 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taataacccc gccccgttga
      541 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga
      601 accgtcagat cctcactctc ttccgcatcg ctgtctgcga gggccagctg ttgggctcgc
      661 ggttgaggac aaactcttcg cggtctttcc agtactcttg gatcggaaac ccgtcggcct
      721 ccgaacggta ctccgccacc gagggacctg agcgagtccg catcgaccgg atcggaaaac
      781 ctctcgagaa aggcgtctaa ccagtcacag tcgcaaggta ggctgagcac cgtggcgggc
      841 ggcagcgggt ggcggtcggg gttgtttctg gcggaggtgc tgctgatgat gtaattaaag
      901 taggcggtct tgagacggcg gatggtcgag gtgaggtgtg gcaggcttga gatccagctg
      961 ttggggtgag tactccctct caaaagcggg cattacttct gcgctaagat tgtcagtttc
     1021 caaaaacgag gaggatttga tattcacctg gcccgatctg gccatacact tgagtgacaa
     1081 tgacatccac tttgcctttc tctccacagg tgtccactcc caggtccaag tttgccacca
     1141 tggagtttca gacccaggta ctcatgtccc tgctgctctg catgtctggt gcggccgccg
     1201 ctgccacagg caatagctct ggcctcgtgt gggtcatcct gaccgacctg agccggacca
     1261 ccatgaccga ggccagcctg agcgaggcca tcaccgcctt tctgaccaac accagcgcca
     1321 cccaccagta cctgaacacc ctgttccggc ctgcccccat catcctggac accggcggca
     1381 atctgctgac cgccctgacc cagatcaccg tgctgctgag caaccccctg accttcccca
     1441 gcgtgctgtt cctggccgag acagccgatg tgtccgccag cgtggtggac atgctgcggc
     1501 agaacgacac caccagcgac atcctgatca tcagcgccca cagcagcaac acccggctgt
     1561 gcgaccccaa gaccaacctg cggaccatgt gcatggtgcc ccgggacatg atcaacgtgc
     1621 ggggcctgct ggaagtggtg tccaaccagc tgaactggta cagcatgagc ctggccctga
     1681 gcaacgacga ctacggcgac ggcgtggccc aggccgtgac atctgagggc cagaccgcca
     1741 gcaccaccgc cacaatcgtg gcccacgcct tcatgaacgg cgctgcctcc accaccgacg
     1801 acgatgccgt ggtggcctcc ctgatcaagt accgggccag aggcgtgggc tgcttcctga
     1861 gagaggccga gacaagacgg ctgcacgccg ccgtggaacg gaatcggaga gccgccaaca
     1921 gcgtgttcct gatcgccagc cgcgagagcc tgaacgtgct gcccgatctg accagcgacg
     1981 tgaacggcac cgccaagccc tggggcgcca tcttcgtgtc cgcctacacc ccccctgccg
     2041 agctggtgtc ccagggcttc tttcccacca cccacggcac agccgtggac gagtacggcg
     2101 ccttcgtgct gagccatctg ctggacggcc tgctgatgct ggatgccgcc aagggcgcca
     2161 tcgacaacat gagcgccctg agggccgcca gattccgggg ctacaccggc aacgtggcct
     2221 tcgacagcat ctactaccag cgggtggaaa ccgtgttcag cctgatcacc gccagccaca
     2281 ccgtgcggaa ccccctggtc acctggaccc tgaagaacta cagcagcgac gccgcccagg
     2341 tcaacgccaa ccccaatgtg accagcgccc tgattaagac cagccccctg cggaccgcca
     2401 agatctgcat ggccagcccc agcaactgcg ccttcaccca gatgatgatc agcatgctgt
     2461 tcgtgctgac agcccacaac gagaacgtgg ccgacaacga cagcgactcc gtgttcaact
     2521 tctaccccgt ggccgtgtcc accggcgcct ctggcgtgat gggactggcc tctctgatcc
     2581 ctatcgcccg gtcctgcacc gtgctgaccg gacctggaag cgacgccgtg gtcatggccg
     2641 tgacccccgt ggtcaacgag ttccacatcc cccagctgga ctacgccgtg tccaacgact
     2701 acttcaccga caacgtgcac acctacccct acttcagcag aagcctgccc aagaacacct
     2761 tcgccgacgt ggccgtgaca cagctgtgcg tgcactacgg ctgggagcgc gtgatcatcg
     2821 tgaccagcaa cgaccagttc ggcatcagca gagcccagag catggccacc cggatgcagc
     2881 agcagaacct gtacgtggaa gccacctact acctgagcga cggcaacaac gccaccgtgg
     2941 ccgactgcat ggaaaagatc tacgccaagc tggtgtctcg gatcatcatc ctgattaacc
     3001 ccttcacagc cgcccaggcc gaaaccttct tcagactgag cgactacctg acctatatgc
     3061 ggcagtacat cttcttcatg gacagcgccc tgtgccacgc cgctgccgcc tctgctaatg
     3121 gcgccgctct gagacagaag ctgcccagca gcatctgcat ctaccccaac gtgacccaga
     3181 cccggctggc cgccctgaat accctgtaca ccagcagcga ctacgccacc acccggcagg
     3241 aaatgcggaa gctgatgtcc gacggcggct tcctgagcag cgtggaaagc tgcgacatca
     3301 gcagcatcag cccctacgcc ggcttcgccg tggatgccgg ctacgtgctg atcgacagcg
     3361 tgtccagagc cgtggccgag aacgtgtccc tgaatgtggc cgcccatctg ctgccctaca
     3421 tcagaagcac cagcatcgac aacttcgccg gcgactgcac catcgacagc accggcaaca
     3481 gactgtacgc cgcctacagc atcaacatcc agctgctgaa cggcagcagc ctgttcatcg
     3541 gcacctggaa cagcaaggcc agccctgccc tggaaatcac cgagtggtcc ttcgtgtggc
     3601 tgaccaacag caccgccgtg cccctggaca ccttccggga cgccagcttt gtgctgagca
     3661 ccgcctttag cgccagcccc ggaggcgcgc ccggcagcct gcaccacatc ctggacgccc
     3721 agaagatgct gtggaaccac agggacagga acctgcctcc tctggcccct ctgggccccc
     3781 accaccatca ccaccattga gtctagacac gtgattaatt aaggatcccc cgacctcgac
     3841 ctctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct
     3901 ctcactcgga aggacatatg ggagggcaaa tcatttggtc gagatccctc ggagatctct
     3961 agctagagga tcgatccccg ccccggacga actaaacctg actacgacat ctctgcccct
     4021 tcttcgcggg gcagtgcatg taatcccttc agttggttgg tacaacttgc caactgggcc
     4081 ctgttccaca tgtgacacgg ggggggacca aacacaaagg ggttctctga ctgtagttga
     4141 catccttata aatggatgtg cacatttgcc aacactgagt ggctttcatc ctggagcaga
     4201 ctttgcagtc tgtggactgc aacacaacat tgcctttatg tgtaactctt ggctgaagct
     4261 cttacaccaa tgctggggga catgtacctc ccaggggccc aggaagacta cgggaggcta
     4321 caccaacgtc aatcagaggg gcctgtgtag ctaccgataa gcggaccctc aagagggcat
     4381 tagcaatagt gtttataagg cccccttgtt aaccctaaac gggtagcata tgcttcccgg
     4441 gtagtagtat atactatcca gactaaccct aattcaatag catatgttac ccaacgggaa
     4501 gcatatgcta tcgaattagg gttagtaaaa gggtcctaag gaacagcgat atctcccacc
     4561 ccatgagctg tcacggtttt atttacatgg ggtcaggatt ccacgagggt agtgaaccat
     4621 tttagtcaca agggcagtgg ctgaagatca aggagcgggc agtgaactct cctgaatctt
     4681 cgcctgcttc ttcattctcc ttcgtttagc taatagaata actgctgagt tgtgaacagt
     4741 aaggtgtatg tgaggtgctc gaaaacaagg tttcaggtga cgcccccaga ataaaatttg
     4801 gacggggggt tcagtggtgg cattgtgcta tgacaccaat ataaccctca caaacccctt
     4861 gggcaataaa tactagtgta ggaatgaaac attctgaata tctttaacaa tagaaatcca
     4921 tggggtgggg acaagccgta aagactggat gtccatctca cacgaattta tggctatggg
     4981 caacacataa tcctagtgca atatgatact ggggttatta agatgtgtcc caggcaggga
     5041 ccaagacagg tgaaccatgt tgttacactc tatttgtaac aaggggaaag agagtggacg
     5101 ccgacagcag cggactccac tggttgtctc taacaccccc gaaaattaaa cggggctcca
     5161 cgccaatggg gcccataaac aaagacaagt ggccactctt ttttttgaaa ttgtggagtg
     5221 ggggcacgcg tcagccccca cacgccgccc tgcggttttg gactgtaaaa taagggtgta
     5281 ataacttggc tgattgtaac cccgctaacc actgcggtca aaccacttgc ccacaaaacc
     5341 actaatggca ccccggggaa tacctgcata agtaggtggg cgggccaaga taggggcgcg
     5401 attgctgcga tctggaggac aaattacaca cacttgcgcc tgagcgccaa gcacagggtt
     5461 gttggtcctc atattcacga ggtcgctgag agcacggtgg gctaatgttg ccatgggtag
     5521 catatactac ccaaatatct ggatagcata tgctatccta atctatatct gggtagcata
     5581 ggctatccta atctatatct gggtagcata tgctatccta atctatatct gggtagtata
     5641 tgctatccta atttatatct gggtagcata ggctatccta atctatatct gggtagcata
     5701 tgctatccta atctatatct gggtagtata tgctatccta atctgtatcc gggtagcata
     5761 tgctatccta atagagatta gggtagtata tgctatccta atttatatct gggtagcata
     5821 tactacccaa atatctggat agcatatgct atcctaatct atatctgggt agcatatgct
     5881 atcctaatct atatctgggt agcataggct atcctaatct atatctgggt agcatatgct
     5941 atcctaatct atatctgggt agtatatgct atcctaattt atatctgggt agcataggct
     6001 atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt agtatatgct
     6061 atcctaatct gtatccgggt agcatatgct atcctcatga taagctgtca aacatgagaa
     6121 ttttcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     6181 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     6241 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     6301 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     6361 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     6421 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     6481 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     6541 aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt
     6601 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     6661 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     6721 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     6781 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     6841 ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa
     6901 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     6961 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     7021 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     7081 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     7141 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     7201 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     7261 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     7321 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     7381 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     7441 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     7501 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     7561 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     7621 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     7681 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     7741 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     7801 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     7861 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     7921 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     7981 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     8041 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     8101 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     8161 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     8221 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     8281 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     8341 aacagctatg accatgatta cgccaagctc tagctagagg tcgaccaatt ctcatgtttg
     8401 acagcttatc atcgcagatc cgggcaacgt tgttgccatt gctgcaggcg cagaactggt
     8461 aggtatggaa gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt
     8521 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata
     8581 tgtacattta tattggctca tgtccaatat gaccgc
//
