LOCUS       sequence-262371-        6342 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 6342)
  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 6342)
  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: D79_Ld_366740
FEATURES             Location/Qualifiers
     source          1..6342
                     /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             1506..1523
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1575..1630
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1629..1648)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3866..3884)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3952..4056
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4057..4125,4126..4917)
                     /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(4275..4294)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5088..5676
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5577..5596
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5830..5847
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5964..5985
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6000..6005,6006..6023,6024..6030)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6038..6054
                     /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     6043..6065
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6062..6078
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6062..6078
                     /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 gactggttgt tagaatggtg caggtcgtgc acagacacgg cgccagatct gccctgatca
     1261 acgacaatac caccgagatc tgcggcacac tgtacccttg tggcgaactg acaggcgaag
     1321 gcgtggaaat ggttcgagcc atcggagagt ctccccatct gcaccctttt gccgtgactc
     1381 aggattggtc tagcctgcct cctctgacag gcgcgcccgg cagcctgcac cacatcctgg
     1441 acgcccagaa gatgctgtgg aaccacaggg acaggaacct gcctcctctg gcccctctgg
     1501 gcccccacca ccatcaccac cattgagtct agacacgtga ttaattaagg atcccccgac
     1561 ctcgacctct ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt
     1621 gtgtctctca ctcggaagga catatgggag ggcaaatcat ttggtcgaga tccctcggag
     1681 atctctagct agaggatcga tccccgcccc ggacgaacta aacctgacta cgacatctct
     1741 gccccttctt cgcggggcag tgcatgtaat cccttcagtt ggttggtaca acttgccaac
     1801 tgggccctgt tccacatgtg acacgggggg ggaccaaaca caaaggggtt ctctgactgt
     1861 agttgacatc cttataaatg gatgtgcaca tttgccaaca ctgagtggct ttcatcctgg
     1921 agcagacttt gcagtctgtg gactgcaaca caacattgcc tttatgtgta actcttggct
     1981 gaagctctta caccaatgct gggggacatg tacctcccag gggcccagga agactacggg
     2041 aggctacacc aacgtcaatc agaggggcct gtgtagctac cgataagcgg accctcaaga
     2101 gggcattagc aatagtgttt ataaggcccc cttgttaacc ctaaacgggt agcatatgct
     2161 tcccgggtag tagtatatac tatccagact aaccctaatt caatagcata tgttacccaa
     2221 cgggaagcat atgctatcga attagggtta gtaaaagggt cctaaggaac agcgatatct
     2281 cccaccccat gagctgtcac ggttttattt acatggggtc aggattccac gagggtagtg
     2341 aaccatttta gtcacaaggg cagtggctga agatcaagga gcgggcagtg aactctcctg
     2401 aatcttcgcc tgcttcttca ttctccttcg tttagctaat agaataactg ctgagttgtg
     2461 aacagtaagg tgtatgtgag gtgctcgaaa acaaggtttc aggtgacgcc cccagaataa
     2521 aatttggacg gggggttcag tggtggcatt gtgctatgac accaatataa ccctcacaaa
     2581 ccccttgggc aataaatact agtgtaggaa tgaaacattc tgaatatctt taacaataga
     2641 aatccatggg gtggggacaa gccgtaaaga ctggatgtcc atctcacacg aatttatggc
     2701 tatgggcaac acataatcct agtgcaatat gatactgggg ttattaagat gtgtcccagg
     2761 cagggaccaa gacaggtgaa ccatgttgtt acactctatt tgtaacaagg ggaaagagag
     2821 tggacgccga cagcagcgga ctccactggt tgtctctaac acccccgaaa attaaacggg
     2881 gctccacgcc aatggggccc ataaacaaag acaagtggcc actctttttt ttgaaattgt
     2941 ggagtggggg cacgcgtcag cccccacacg ccgccctgcg gttttggact gtaaaataag
     3001 ggtgtaataa cttggctgat tgtaaccccg ctaaccactg cggtcaaacc acttgcccac
     3061 aaaaccacta atggcacccc ggggaatacc tgcataagta ggtgggcggg ccaagatagg
     3121 ggcgcgattg ctgcgatctg gaggacaaat tacacacact tgcgcctgag cgccaagcac
     3181 agggttgttg gtcctcatat tcacgaggtc gctgagagca cggtgggcta atgttgccat
     3241 gggtagcata tactacccaa atatctggat agcatatgct atcctaatct atatctgggt
     3301 agcataggct atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt
     3361 agtatatgct atcctaattt atatctgggt agcataggct atcctaatct atatctgggt
     3421 agcatatgct atcctaatct atatctgggt agtatatgct atcctaatct gtatccgggt
     3481 agcatatgct atcctaatag agattagggt agtatatgct atcctaattt atatctgggt
     3541 agcatatact acccaaatat ctggatagca tatgctatcc taatctatat ctgggtagca
     3601 tatgctatcc taatctatat ctgggtagca taggctatcc taatctatat ctgggtagca
     3661 tatgctatcc taatctatat ctgggtagta tatgctatcc taatttatat ctgggtagca
     3721 taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagta
     3781 tatgctatcc taatctgtat ccgggtagca tatgctatcc tcatgataag ctgtcaaaca
     3841 tgagaatttt cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     3901 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     3961 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4021 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     4081 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     4141 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     4201 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     4261 acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa
     4321 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     4381 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     4441 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     4501 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     4561 gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg
     4621 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     4681 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     4741 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     4801 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     4861 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     4921 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     4981 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5041 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     5101 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     5161 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     5221 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     5281 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     5341 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     5401 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     5461 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     5521 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     5581 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     5641 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     5701 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     5761 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     5821 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     5881 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     5941 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     6001 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     6061 acaggaaaca gctatgacca tgattacgcc aagctctagc tagaggtcga ccaattctca
     6121 tgtttgacag cttatcatcg cagatccggg caacgttgtt gccattgctg caggcgcaga
     6181 actggtaggt atggaagatc tatacattga atcaatattg gcaattagcc atattagtca
     6241 ttggttatat agcataaatc aatattggct attggccatt gcatacgttg tatctatatc
     6301 ataatatgta catttatatt ggctcatgtc caatatgacc gc
//
