LOCUS       sequence-262326-        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"
     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             2796..2813
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2865..2920
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2919..2938)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5156..5174)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5242..5346
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5347..5415,5416..6207)
                     /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(5565..5584)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6378..6966
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6867..6886
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7120..7137
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7254..7275
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7290..7295,7296..7313,7314..7320)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7328..7344
                     /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     7333..7355
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7352..7368
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7352..7368
                     /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 ccggacctgt gctggaagtg ggagtggacc gggccatcac cttcaacaga cagctgggcg
     1261 acggcgacga gctgctgttc ggaagcagag ccgtgctgct gcggggcgag atcgtgcggt
     1321 atacagccgg cgcttccgcc agctaccacg gctacaccgt gtacgtgcag tgcagcgagc
     1381 acctgaccaa cttcgccctg cagcagctgg ccgctcccga gctgtctaag gccgccaagg
     1441 gcctggtggt ggaactgtgc gacaccgaca ccaccaacga ggacgtgctg agcttcttct
     1501 tctccaacgc cgctgccagc atccccgtgt actttctgcc caacggcacc gccagcaaag
     1561 agctgagcag actgctgagc gtggccgccc agagaagcgc caccgagagg gtggtgctgt
     1621 ccgtggccaa gaccaggcgg ctgagcgagc tggtggaaaa cagcaccctg ccctccgcca
     1681 ccatcgagag ccactacgtg cacaagccca agcagaccag aagcaaagcc gccagcgctg
     1741 ccgccgctcc tcaggtgctg gtcaccgccc acttcgactc cctgggcgtg tccccagcca
     1801 gcagaacctc tggcggaacc agcggagccg tggtggccat ggaactgtgg cggagactga
     1861 ccagcacccc ctacgccaga caggaaagcg tcgcccctta cgaagtgacc gtcctgctgg
     1921 gcagcaccag ccggttcaac tacgccggca ccacaagctg gctgagccag cacaccgata
     1981 gagagctgga ccagttccgg gtggtcctgt gcctggacga actgctgccc cccagagaga
     2041 caagcgagaa cgcccccgac ctgtatctgc atgtgcagga tgtgctgatg aagcggcagc
     2101 acggccagca ggtcgtggaa caggtggaag ccgccgcaaa gctgctggga atcagcctga
     2161 aggtggtgtc cgccaagaca aactaccagc actacgacct ggaattcgag cacgaggcct
     2221 tcgccagccg gcagatgacc gccatgaccc tgagcaccca ccgggtgcac cataacgacc
     2281 agctgttccg ggacctgcgg aggcctcctg ccacagccgc tgatgccgct atgctggcca
     2341 agcgggccga cctggtcgag gccattgtgc ggattgtggc cgaggctgcc cctagcgccg
     2401 agggcgtgac aacagtgtgg cctggcgccg ctagctacat ccagggcatg ctgcactacg
     2461 cctccgacag ccagagatcc cccgtggccc agaacggcgc tggcctgaga cagtacgccg
     2521 ccacagtgga acaccacatg agagcccagg ccgctgctgc tcagagagcc gcctttacca
     2581 gcgccagctc tgtggccacc taccaggaac tgcggacccc cggcatcacc ctgttcggcc
     2641 catacgaaga gacaatgcag gtgttcatgg ccaagtccta cctgttcgag tgcgccgtgg
     2701 gcgcgcccgg cagcctgcac cacatcctgg acgcccagaa gatgctgtgg aaccacaggg
     2761 acaggaacct gcctcctctg gcccctctgg gcccccacca ccatcaccac cattgagtct
     2821 agacacgtga ttaattaagg atcccccgac ctcgacctct ggctaataaa ggaaatttat
     2881 tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag
     2941 ggcaaatcat ttggtcgaga tccctcggag atctctagct agaggatcga tccccgcccc
     3001 ggacgaacta aacctgacta cgacatctct gccccttctt cgcggggcag tgcatgtaat
     3061 cccttcagtt ggttggtaca acttgccaac tgggccctgt tccacatgtg acacgggggg
     3121 ggaccaaaca caaaggggtt ctctgactgt agttgacatc cttataaatg gatgtgcaca
     3181 tttgccaaca ctgagtggct ttcatcctgg agcagacttt gcagtctgtg gactgcaaca
     3241 caacattgcc tttatgtgta actcttggct gaagctctta caccaatgct gggggacatg
     3301 tacctcccag gggcccagga agactacggg aggctacacc aacgtcaatc agaggggcct
     3361 gtgtagctac cgataagcgg accctcaaga gggcattagc aatagtgttt ataaggcccc
     3421 cttgttaacc ctaaacgggt agcatatgct tcccgggtag tagtatatac tatccagact
     3481 aaccctaatt caatagcata tgttacccaa cgggaagcat atgctatcga attagggtta
     3541 gtaaaagggt cctaaggaac agcgatatct cccaccccat gagctgtcac ggttttattt
     3601 acatggggtc aggattccac gagggtagtg aaccatttta gtcacaaggg cagtggctga
     3661 agatcaagga gcgggcagtg aactctcctg aatcttcgcc tgcttcttca ttctccttcg
     3721 tttagctaat agaataactg ctgagttgtg aacagtaagg tgtatgtgag gtgctcgaaa
     3781 acaaggtttc aggtgacgcc cccagaataa aatttggacg gggggttcag tggtggcatt
     3841 gtgctatgac accaatataa ccctcacaaa ccccttgggc aataaatact agtgtaggaa
     3901 tgaaacattc tgaatatctt taacaataga aatccatggg gtggggacaa gccgtaaaga
     3961 ctggatgtcc atctcacacg aatttatggc tatgggcaac acataatcct agtgcaatat
     4021 gatactgggg ttattaagat gtgtcccagg cagggaccaa gacaggtgaa ccatgttgtt
     4081 acactctatt tgtaacaagg ggaaagagag tggacgccga cagcagcgga ctccactggt
     4141 tgtctctaac acccccgaaa attaaacggg gctccacgcc aatggggccc ataaacaaag
     4201 acaagtggcc actctttttt ttgaaattgt ggagtggggg cacgcgtcag cccccacacg
     4261 ccgccctgcg gttttggact gtaaaataag ggtgtaataa cttggctgat tgtaaccccg
     4321 ctaaccactg cggtcaaacc acttgcccac aaaaccacta atggcacccc ggggaatacc
     4381 tgcataagta ggtgggcggg ccaagatagg ggcgcgattg ctgcgatctg gaggacaaat
     4441 tacacacact tgcgcctgag cgccaagcac agggttgttg gtcctcatat tcacgaggtc
     4501 gctgagagca cggtgggcta atgttgccat gggtagcata tactacccaa atatctggat
     4561 agcatatgct atcctaatct atatctgggt agcataggct atcctaatct atatctgggt
     4621 agcatatgct atcctaatct atatctgggt agtatatgct atcctaattt atatctgggt
     4681 agcataggct atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt
     4741 agtatatgct atcctaatct gtatccgggt agcatatgct atcctaatag agattagggt
     4801 agtatatgct atcctaattt atatctgggt agcatatact acccaaatat ctggatagca
     4861 tatgctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagca
     4921 taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagta
     4981 tatgctatcc taatttatat ctgggtagca taggctatcc taatctatat ctgggtagca
     5041 tatgctatcc taatctatat ctgggtagta tatgctatcc taatctgtat ccgggtagca
     5101 tatgctatcc tcatgataag ctgtcaaaca tgagaatttt cttgaagacg aaagggcctc
     5161 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     5221 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     5281 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     5341 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     5401 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     5461 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     5521 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     5581 ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     5641 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     5701 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     5761 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     5821 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     5881 acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     5941 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     6001 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     6061 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     6121 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     6181 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     6241 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     6301 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     6361 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     6421 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     6481 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     6541 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     6601 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     6661 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     6721 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     6781 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     6841 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     6901 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     6961 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     7021 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     7081 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     7141 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     7201 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     7261 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     7321 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     7381 aagctctagc tagaggtcga ccaattctca tgtttgacag cttatcatcg cagatccggg
     7441 caacgttgtt gccattgctg caggcgcaga actggtaggt atggaagatc tatacattga
     7501 atcaatattg gcaattagcc atattagtca ttggttatat agcataaatc aatattggct
     7561 attggccatt gcatacgttg tatctatatc ataatatgta catttatatt ggctcatgtc
     7621 caatatgacc gc
//
