LOCUS       sequence-274898-        6852 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 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 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: D39_Ld_360680
FEATURES             Location/Qualifiers
     source          1..6852
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     1086..1105
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1128..1137
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2010..2027
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2079..2134
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2133..2152)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4370..4388)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4456..4560
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4561..4629,4630..5421)
                     /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(4779..4798)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5592..6180
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6081..6100
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6334..6351
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6468..6489
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6504..6509,6510..6527,6528..6534)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6542..6558
                     /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     6547..6569
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6566..6582
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6566..6582
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agatcctcac tctcttccgc atcgctgtct gcgagggcca gctgttgggc tcgcggttga
      661 ggacaaactc ttcgcggtct ttccagtact cttggatcgg aaacccgtcg gcctccgaac
      721 ggtactccgc caccgaggga cctgagcgag tccgcatcga ccggatcgga aaacctctcg
      781 agaaaggcgt ctaaccagtc acagtcgcaa ggtaggctga gcaccgtggc gggcggcagc
      841 gggtggcggt cggggttgtt tctggcggag gtgctgctga tgatgtaatt aaagtaggcg
      901 gtcttgagac ggcggatggt cgaggtgagg tgtggcaggc ttgagatcca gctgttgggg
      961 tgagtactcc ctctcaaaag cgggcattac ttctgcgcta agattgtcag tttccaaaaa
     1021 cgaggaggat ttgatattca cctggcccga tctggccata cacttgagtg acaatgacat
     1081 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caagtttgcc accatggagt
     1141 ttcagaccca ggtactcatg tccctgctgc tctgcatgtc tggtgcggcc gccaccgtgc
     1201 ctctgagcag agtgtttgtg cacgccggcc acaacgtggg cgtgcgggtg taccacgtgc
     1261 ccttcatcgt gctgttcagc ctgaagctgc cccggctgtg ctgcggctgc accttcaatc
     1321 ctccacccgc cgagagcgac accctgcctg ccatcccact gagcaccagc ccctggacct
     1381 acgccaacaa gtggcagcgg aacaccaaga cccggctgcc ctgctgcccc ctgccatacc
     1441 aggaacagtg gtggtccctg agcctgagag tgtccagccg gctgagcccc cccatgtacc
     1501 tgcctctgtc caccgcccct gacaccgtgc tgtccccccc aacctccccc agacccaaga
     1561 gaacagaggc cgtggccagc atccgggcct gcagaagcac ccacagctac cgggccatct
     1621 tccggcaccg gtttctgtgg ttccacctga gaaggctgag cgtgctgcag taccggggca
     1681 gcctgagccc tatgatgctg cggagaagcc tgatcgccct gaactggcgg gaccagggca
     1741 tcagctacgt gaagtacctg aacgtgacca ccgaggccct gcacatggcc accaaggaca
     1801 aggccagagc caagtacagc cggttcagcg cccccaacta cgtggccatg aagaacgact
     1861 ccaccggcgc catggaagaa gtgaagaagg tccccgcctt cacaaaggac tatggcgcgc
     1921 ccggcagcct gcaccacatc ctggacgccc agaagatgct gtggaaccac agggacagga
     1981 acctgcctcc tctggcccct ctgggccccc accaccatca ccaccattga gtctagacac
     2041 gtgattaatt aaggatcccc cgacctcgac ctctggctaa taaaggaaat ttattttcat
     2101 tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa
     2161 tcatttggtc gagatccctc ggagatctct agctagagga tcgatccccg ccccggacga
     2221 actaaacctg actacgacat ctctgcccct tcttcgcggg gcagtgcatg taatcccttc
     2281 agttggttgg tacaacttgc caactgggcc ctgttccaca tgtgacacgg ggggggacca
     2341 aacacaaagg ggttctctga ctgtagttga catccttata aatggatgtg cacatttgcc
     2401 aacactgagt ggctttcatc ctggagcaga ctttgcagtc tgtggactgc aacacaacat
     2461 tgcctttatg tgtaactctt ggctgaagct cttacaccaa tgctggggga catgtacctc
     2521 ccaggggccc aggaagacta cgggaggcta caccaacgtc aatcagaggg gcctgtgtag
     2581 ctaccgataa gcggaccctc aagagggcat tagcaatagt gtttataagg cccccttgtt
     2641 aaccctaaac gggtagcata tgcttcccgg gtagtagtat atactatcca gactaaccct
     2701 aattcaatag catatgttac ccaacgggaa gcatatgcta tcgaattagg gttagtaaaa
     2761 gggtcctaag gaacagcgat atctcccacc ccatgagctg tcacggtttt atttacatgg
     2821 ggtcaggatt ccacgagggt agtgaaccat tttagtcaca agggcagtgg ctgaagatca
     2881 aggagcgggc agtgaactct cctgaatctt cgcctgcttc ttcattctcc ttcgtttagc
     2941 taatagaata actgctgagt tgtgaacagt aaggtgtatg tgaggtgctc gaaaacaagg
     3001 tttcaggtga cgcccccaga ataaaatttg gacggggggt tcagtggtgg cattgtgcta
     3061 tgacaccaat ataaccctca caaacccctt gggcaataaa tactagtgta ggaatgaaac
     3121 attctgaata tctttaacaa tagaaatcca tggggtgggg acaagccgta aagactggat
     3181 gtccatctca cacgaattta tggctatggg caacacataa tcctagtgca atatgatact
     3241 ggggttatta agatgtgtcc caggcaggga ccaagacagg tgaaccatgt tgttacactc
     3301 tatttgtaac aaggggaaag agagtggacg ccgacagcag cggactccac tggttgtctc
     3361 taacaccccc gaaaattaaa cggggctcca cgccaatggg gcccataaac aaagacaagt
     3421 ggccactctt ttttttgaaa ttgtggagtg ggggcacgcg tcagccccca cacgccgccc
     3481 tgcggttttg gactgtaaaa taagggtgta ataacttggc tgattgtaac cccgctaacc
     3541 actgcggtca aaccacttgc ccacaaaacc actaatggca ccccggggaa tacctgcata
     3601 agtaggtggg cgggccaaga taggggcgcg attgctgcga tctggaggac aaattacaca
     3661 cacttgcgcc tgagcgccaa gcacagggtt gttggtcctc atattcacga ggtcgctgag
     3721 agcacggtgg gctaatgttg ccatgggtag catatactac ccaaatatct ggatagcata
     3781 tgctatccta atctatatct gggtagcata ggctatccta atctatatct gggtagcata
     3841 tgctatccta atctatatct gggtagtata tgctatccta atttatatct gggtagcata
     3901 ggctatccta atctatatct gggtagcata tgctatccta atctatatct gggtagtata
     3961 tgctatccta atctgtatcc gggtagcata tgctatccta atagagatta gggtagtata
     4021 tgctatccta atttatatct gggtagcata tactacccaa atatctggat agcatatgct
     4081 atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt agcataggct
     4141 atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt agtatatgct
     4201 atcctaattt atatctgggt agcataggct atcctaatct atatctgggt agcatatgct
     4261 atcctaatct atatctgggt agtatatgct atcctaatct gtatccgggt agcatatgct
     4321 atcctcatga taagctgtca aacatgagaa ttttcttgaa gacgaaaggg cctcgtgata
     4381 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     4441 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     4501 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     4561 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     4621 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     4681 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     4741 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     4801 cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     4861 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     4921 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     4981 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     5041 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     5101 cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     5161 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     5221 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     5281 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     5341 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     5401 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     5461 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     5521 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     5581 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     5641 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     5701 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     5761 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     5821 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     5881 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     5941 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     6001 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     6061 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     6121 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     6181 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     6241 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     6301 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     6361 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     6421 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     6481 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     6541 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagctc
     6601 tagctagagg tcgaccaatt ctcatgtttg acagcttatc atcgcagatc cgggcaacgt
     6661 tgttgccatt gctgcaggcg cagaactggt aggtatggaa gatctataca ttgaatcaat
     6721 attggcaatt agccatatta gtcattggtt atatagcata aatcaatatt ggctattggc
     6781 cattgcatac gttgtatcta tatcataata tgtacattta tattggctca tgtccaatat
     6841 gaccgccatg tt
//
