LOCUS       sequence-262385-        7272 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 7272)
  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 7272)
  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: D93_Ld_361080
FEATURES             Location/Qualifiers
     source          1..7272
                     /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             2436..2453
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2505..2560
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2559..2578)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4796..4814)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4882..4986
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4987..5055,5056..5847)
                     /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(5205..5224)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6018..6606
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6507..6526
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6760..6777
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6894..6915
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6930..6935,6936..6953,6954..6960)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6968..6984
                     /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     6973..6995
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6992..7008
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6992..7008
                     /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 atactgctca tcctcaccat tctaggtccc atacagtcac aatagagtcc ggaacactgc
     1261 ccgaagcaaa ggcttttgcg cccttgaggc tgcgggccgg ggaatgggta tataacttca
     1321 atccaggcga agaagcattc ttctttgagc ccgaggatgc cggtaacagg cagtggcagc
     1381 agtggaaggc tgatctcagg ttgttgcagg cctatcagga gaaacagcga ctttccaact
     1441 atccctctgg cggggagaag actgacaccc ccgataagcc tggaaatact gagcgcagag
     1501 agccgggcgc gttgcctatt gccgagtatt tctccgaccg caacgttcag aggcggtatg
     1561 ctgaacgcct gaaggaacag tcttttcata acctttccga cgcacagggt gctgcagatg
     1621 ctcgcgccct ggtagaaggc gagatagctg caatgtggca tctcgtcaga agtgctggca
     1681 gttcagatag ccttaatgat cagcagtttc tgggactggt ggaaaggcat ttcagtcata
     1741 ctatagctaa tacctcagca gctgccgcag ccgaagccac aacaccttca agctttccct
     1801 tcgccttgct tcgcgttccc ctggtactcg aggcggacgt tctgccattt atggagagag
     1861 aaatggcgca tgtgagggat ttgtggaagt caaggagagc cggtgccgcc acccccgcag
     1921 ccagctcagt ggaagtggag tctcagcttc agcggctgtc ttctgctgtg gagaactcaa
     1981 agactgttgg caccgagctc gctgctctgg tggagccttt ggctgatgca gctatcggat
     2041 gggtagctag cactctgaca accacacgtg aagcgatagt agcaaacgat gaaagcaccg
     2101 aagccaagcg cagcgagctg aaacagcttc tgaaaatgca gtctcgtctc agaaaagtga
     2161 gaccaatact tgagaaacat cgcaaagttt atcagtccgt tctccaaagg ggccagctgc
     2221 ccaatgccca agcaaccctc tttgctcttc aacttatagc caagcaacga ttcggggacg
     2281 acgtttctgg ccttgtggaa gcggtacctc tcctctccac gactgcgccc ctgaccggcg
     2341 gcgcgcccgg cagcctgcac cacatcctgg acgcccagaa gatgctgtgg aaccacaggg
     2401 acaggaacct gcctcctctg gcccctctgg gcccccacca ccatcaccac cattgagtct
     2461 agacacgtga ttaattaagg atcccccgac ctcgacctct ggctaataaa ggaaatttat
     2521 tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag
     2581 ggcaaatcat ttggtcgaga tccctcggag atctctagct agaggatcga tccccgcccc
     2641 ggacgaacta aacctgacta cgacatctct gccccttctt cgcggggcag tgcatgtaat
     2701 cccttcagtt ggttggtaca acttgccaac tgggccctgt tccacatgtg acacgggggg
     2761 ggaccaaaca caaaggggtt ctctgactgt agttgacatc cttataaatg gatgtgcaca
     2821 tttgccaaca ctgagtggct ttcatcctgg agcagacttt gcagtctgtg gactgcaaca
     2881 caacattgcc tttatgtgta actcttggct gaagctctta caccaatgct gggggacatg
     2941 tacctcccag gggcccagga agactacggg aggctacacc aacgtcaatc agaggggcct
     3001 gtgtagctac cgataagcgg accctcaaga gggcattagc aatagtgttt ataaggcccc
     3061 cttgttaacc ctaaacgggt agcatatgct tcccgggtag tagtatatac tatccagact
     3121 aaccctaatt caatagcata tgttacccaa cgggaagcat atgctatcga attagggtta
     3181 gtaaaagggt cctaaggaac agcgatatct cccaccccat gagctgtcac ggttttattt
     3241 acatggggtc aggattccac gagggtagtg aaccatttta gtcacaaggg cagtggctga
     3301 agatcaagga gcgggcagtg aactctcctg aatcttcgcc tgcttcttca ttctccttcg
     3361 tttagctaat agaataactg ctgagttgtg aacagtaagg tgtatgtgag gtgctcgaaa
     3421 acaaggtttc aggtgacgcc cccagaataa aatttggacg gggggttcag tggtggcatt
     3481 gtgctatgac accaatataa ccctcacaaa ccccttgggc aataaatact agtgtaggaa
     3541 tgaaacattc tgaatatctt taacaataga aatccatggg gtggggacaa gccgtaaaga
     3601 ctggatgtcc atctcacacg aatttatggc tatgggcaac acataatcct agtgcaatat
     3661 gatactgggg ttattaagat gtgtcccagg cagggaccaa gacaggtgaa ccatgttgtt
     3721 acactctatt tgtaacaagg ggaaagagag tggacgccga cagcagcgga ctccactggt
     3781 tgtctctaac acccccgaaa attaaacggg gctccacgcc aatggggccc ataaacaaag
     3841 acaagtggcc actctttttt ttgaaattgt ggagtggggg cacgcgtcag cccccacacg
     3901 ccgccctgcg gttttggact gtaaaataag ggtgtaataa cttggctgat tgtaaccccg
     3961 ctaaccactg cggtcaaacc acttgcccac aaaaccacta atggcacccc ggggaatacc
     4021 tgcataagta ggtgggcggg ccaagatagg ggcgcgattg ctgcgatctg gaggacaaat
     4081 tacacacact tgcgcctgag cgccaagcac agggttgttg gtcctcatat tcacgaggtc
     4141 gctgagagca cggtgggcta atgttgccat gggtagcata tactacccaa atatctggat
     4201 agcatatgct atcctaatct atatctgggt agcataggct atcctaatct atatctgggt
     4261 agcatatgct atcctaatct atatctgggt agtatatgct atcctaattt atatctgggt
     4321 agcataggct atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt
     4381 agtatatgct atcctaatct gtatccgggt agcatatgct atcctaatag agattagggt
     4441 agtatatgct atcctaattt atatctgggt agcatatact acccaaatat ctggatagca
     4501 tatgctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagca
     4561 taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagta
     4621 tatgctatcc taatttatat ctgggtagca taggctatcc taatctatat ctgggtagca
     4681 tatgctatcc taatctatat ctgggtagta tatgctatcc taatctgtat ccgggtagca
     4741 tatgctatcc tcatgataag ctgtcaaaca tgagaatttt cttgaagacg aaagggcctc
     4801 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     4861 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     4921 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     4981 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     5041 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     5101 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     5161 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     5221 ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     5281 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     5341 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     5401 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     5461 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     5521 acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     5581 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     5641 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     5701 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     5761 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     5821 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     5881 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     5941 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     6001 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     6061 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     6121 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     6181 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     6241 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     6301 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     6361 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     6421 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     6481 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     6541 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     6601 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     6661 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     6721 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     6781 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     6841 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     6901 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     6961 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     7021 aagctctagc tagaggtcga ccaattctca tgtttgacag cttatcatcg cagatccggg
     7081 caacgttgtt gccattgctg caggcgcaga actggtaggt atggaagatc tatacattga
     7141 atcaatattg gcaattagcc atattagtca ttggttatat agcataaatc aatattggct
     7201 attggccatt gcatacgttg tatctatatc ataatatgta catttatatt ggctcatgtc
     7261 caatatgacc gc
//
