LOCUS       sequence-262312-        7656 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 7656)
  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 7656)
  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: D17_Ld_090870
FEATURES             Location/Qualifiers
     source          1..7656
                     /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             2820..2837
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2889..2944
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2943..2962)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5180..5198)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5266..5370
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5371..5439,5440..6231)
                     /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(5589..5608)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6402..6990
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6891..6910
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7144..7161
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7278..7299
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7314..7319,7320..7337,7338..7344)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7352..7368
                     /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     7357..7379
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7376..7392
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7376..7392
                     /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 gcggccgcca
     1201 ccggcagacc taagcagcct atcagcaagg gcaccaccgc cggcacaagc agccacgcca
     1261 gcagcagcct gtgccccaga tgccccctgt gcagcagaag cggcaccccc atcaaagagc
     1321 ggctgaagta ctgccacacc accagcggct acatcatcag cctgttcagc tggggcacct
     1381 ctgccctggc cggcaagcag agaaccggcg tgtccaacag agagcagcgg cagctgtacc
     1441 ccctggaact gcccttcgcc accgacgacg ccgtgcggaa gcagtgcatg cgggacatgc
     1501 aggacctgct gctggccaga gtgcggggcc agcctcaggt ggtcatgccc ctgctggacg
     1561 tgctgcggag aaagctggcc tacccccgcg agcctgtggt cgtgcacctg gctggcgaca
     1621 acggcgtggg caagacccac accgccagac tggtgtccca ggccctgagc ctgagatgcg
     1681 ccccagacag agatgtgtgc gacgccggcg acaatctgct gaccattgcc ggcaccggct
     1741 tcgacggcct gtctgtggcc gaggcccggc agagaatcat cggccagatc atcacccaca
     1801 cagacagata cccccacggg gtggtgctga tcgacgacct gaccgccatg gaccccagcc
     1861 tcgtgtctgc tctggccccc ctgtttggca gagccgccca cttccccgag cagctgacca
     1921 gacctagcac cgccacagcc gctggcgcca gacacgccgc tgccaagagc ctgcagacca
     1981 tgcacgagag caccaagggc ctgggccctg atggcggaag aggctctgca gccaagaagg
     2041 gcgaggacga gaagtgggag gctgccgctg gaacaggcgg agcccagcca agcacaagcg
     2101 agatggccag ctccagcctg tcctccggcc agccacctcc tccactgagc cagctgatca
     2161 tcttcgtgac caccgacttc ggccggcagg gccggaccgt gggaaagagc agaaccgaga
     2221 tcgaggccat gatccagcac gacttcgccg acctgtacgg caccctgctg cccgcctaca
     2281 cccggacctt cgtgtactac cccttcacca gcgccatggc tgaggatgtc gtgcggagcg
     2341 ccgtgaccga tctgccttgt gccctgggcg agagactgat cgcctccagc accatctccg
     2401 atgaggccgt gtccttcctg gtgcagcagc acagacagct gtggtccggc aaagagaacg
     2461 gccacgccct gcggcggctg gtggaagatg agatcgtgtc tcagctggtg gtgtactggg
     2521 agacacacgc ccagcagaaa cacttcgaga ggctgcgggt gcactttgag ctggacgagg
     2581 ccagcatgag agtgctgctg aggacccccc acaccagccg gacaaccgcc tctcagcctc
     2641 ctgccgccct gtccagcgtg gaagccagag cctatggcca cggcggagct gacgatggcg
     2701 acggcgagga aagaagcgat ctgggcgcgc ccggcagcct gcaccacatc ctggacgccc
     2761 agaagatgct gtggaaccac agggacagga acctgcctcc tctggcccct ctgggccccc
     2821 accaccatca ccaccattga gtctagacac gtgattaatt aaggatcccc cgacctcgac
     2881 ctctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct
     2941 ctcactcgga aggacatatg ggagggcaaa tcatttggtc gagatccctc ggagatctct
     3001 agctagagga tcgatccccg ccccggacga actaaacctg actacgacat ctctgcccct
     3061 tcttcgcggg gcagtgcatg taatcccttc agttggttgg tacaacttgc caactgggcc
     3121 ctgttccaca tgtgacacgg ggggggacca aacacaaagg ggttctctga ctgtagttga
     3181 catccttata aatggatgtg cacatttgcc aacactgagt ggctttcatc ctggagcaga
     3241 ctttgcagtc tgtggactgc aacacaacat tgcctttatg tgtaactctt ggctgaagct
     3301 cttacaccaa tgctggggga catgtacctc ccaggggccc aggaagacta cgggaggcta
     3361 caccaacgtc aatcagaggg gcctgtgtag ctaccgataa gcggaccctc aagagggcat
     3421 tagcaatagt gtttataagg cccccttgtt aaccctaaac gggtagcata tgcttcccgg
     3481 gtagtagtat atactatcca gactaaccct aattcaatag catatgttac ccaacgggaa
     3541 gcatatgcta tcgaattagg gttagtaaaa gggtcctaag gaacagcgat atctcccacc
     3601 ccatgagctg tcacggtttt atttacatgg ggtcaggatt ccacgagggt agtgaaccat
     3661 tttagtcaca agggcagtgg ctgaagatca aggagcgggc agtgaactct cctgaatctt
     3721 cgcctgcttc ttcattctcc ttcgtttagc taatagaata actgctgagt tgtgaacagt
     3781 aaggtgtatg tgaggtgctc gaaaacaagg tttcaggtga cgcccccaga ataaaatttg
     3841 gacggggggt tcagtggtgg cattgtgcta tgacaccaat ataaccctca caaacccctt
     3901 gggcaataaa tactagtgta ggaatgaaac attctgaata tctttaacaa tagaaatcca
     3961 tggggtgggg acaagccgta aagactggat gtccatctca cacgaattta tggctatggg
     4021 caacacataa tcctagtgca atatgatact ggggttatta agatgtgtcc caggcaggga
     4081 ccaagacagg tgaaccatgt tgttacactc tatttgtaac aaggggaaag agagtggacg
     4141 ccgacagcag cggactccac tggttgtctc taacaccccc gaaaattaaa cggggctcca
     4201 cgccaatggg gcccataaac aaagacaagt ggccactctt ttttttgaaa ttgtggagtg
     4261 ggggcacgcg tcagccccca cacgccgccc tgcggttttg gactgtaaaa taagggtgta
     4321 ataacttggc tgattgtaac cccgctaacc actgcggtca aaccacttgc ccacaaaacc
     4381 actaatggca ccccggggaa tacctgcata agtaggtggg cgggccaaga taggggcgcg
     4441 attgctgcga tctggaggac aaattacaca cacttgcgcc tgagcgccaa gcacagggtt
     4501 gttggtcctc atattcacga ggtcgctgag agcacggtgg gctaatgttg ccatgggtag
     4561 catatactac ccaaatatct ggatagcata tgctatccta atctatatct gggtagcata
     4621 ggctatccta atctatatct gggtagcata tgctatccta atctatatct gggtagtata
     4681 tgctatccta atttatatct gggtagcata ggctatccta atctatatct gggtagcata
     4741 tgctatccta atctatatct gggtagtata tgctatccta atctgtatcc gggtagcata
     4801 tgctatccta atagagatta gggtagtata tgctatccta atttatatct gggtagcata
     4861 tactacccaa atatctggat agcatatgct atcctaatct atatctgggt agcatatgct
     4921 atcctaatct atatctgggt agcataggct atcctaatct atatctgggt agcatatgct
     4981 atcctaatct atatctgggt agtatatgct atcctaattt atatctgggt agcataggct
     5041 atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt agtatatgct
     5101 atcctaatct gtatccgggt agcatatgct atcctcatga taagctgtca aacatgagaa
     5161 ttttcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     5221 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     5281 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     5341 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     5401 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     5461 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     5521 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     5581 aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt
     5641 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     5701 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5761 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     5821 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5881 ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa
     5941 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     6001 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     6061 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     6121 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     6181 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     6241 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     6301 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     6361 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     6421 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     6481 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     6541 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6601 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6661 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6721 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6781 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6841 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6901 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6961 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     7021 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     7081 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     7141 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     7201 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     7261 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     7321 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     7381 aacagctatg accatgatta cgccaagctc tagctagagg tcgaccaatt ctcatgtttg
     7441 acagcttatc atcgcagatc cgggcaacgt tgttgccatt gctgcaggcg cagaactggt
     7501 aggtatggaa gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt
     7561 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata
     7621 tgtacattta tattggctca tgtccaatat gaccgc
//
