LOCUS       sequence-262377-        7062 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 7062)
  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 7062)
  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: D85_Ld_120350
FEATURES             Location/Qualifiers
     source          1..7062
                     /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             2226..2243
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2295..2350
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2349..2368)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4586..4604)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4672..4776
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4777..4845,4846..5637)
                     /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(4995..5014)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5808..6396
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6297..6316
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6550..6567
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6684..6705
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6720..6725,6726..6743,6744..6750)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6758..6774
                     /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     6763..6785
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6782..6798
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6782..6798
                     /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 gcggccgcct
     1201 ggtgggataa agggcacatg tgtattgccg aaattgctag acgtaacttg aagcctaacg
     1261 tgcagtccaa agtgcaggct tgtgccgatg tactcaataa gattggacct tttcctaagt
     1321 ctactaacat cgtggagctt ggcccctggg ctgatgatct caaaagcatg ggcctctaca
     1381 ccatgagtac atggcacttt attgatacaa tttacaaccc gcaggacgtt aaggtaacca
     1441 tcaatccagt ggagattgta aatgtggcgt ctgtgatccc aatgctcatt agcgcgataa
     1501 cctctcctgc tgcaaccagt gatattatta ccactagtgt cgcaaacctg atacactttg
     1561 tgggagacat tcatatgcct ttgcatagtg ccgacctgtt cagccctgaa taccctttgg
     1621 gtgacttggg tggaaacaag cagatcgtca tagtaaacga atccgcaggc acctctatga
     1681 aattgcatgc cttttgggac agcatgtgtg aggggcctca aaacaatgct gtaaggcccc
     1741 tggacaaaga cgcctacgca gagctgtcag cctttgtgga caatctggtg aagtcataca
     1801 gtttcactga ggaacagatg atgatgacta acagtaccat tatggccgcc gagtcctatg
     1861 aattggccgt gaagaacgtc tacccgggta tatctgatag gacagtgctt tccgaaactt
     1921 acaaggccaa cgggaagatt ctggcagccg ggcgggtcac tctggcaggc tacagactgg
     1981 ctacaattct gaatacagct ctggctgggg tgtcattgga tacaatcatg aacgggacaa
     2041 agcacatgca ggatgaggtt gaggtgaccc atacaggcga cacctataac tactacgctt
     2101 tctcaggcgt cgagtctgga gcagccgccg gcgcgcccgg cagcctgcac cacatcctgg
     2161 acgcccagaa gatgctgtgg aaccacaggg acaggaacct gcctcctctg gcccctctgg
     2221 gcccccacca ccatcaccac cattgagtct agacacgtga ttaattaagg atcccccgac
     2281 ctcgacctct ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt
     2341 gtgtctctca ctcggaagga catatgggag ggcaaatcat ttggtcgaga tccctcggag
     2401 atctctagct agaggatcga tccccgcccc ggacgaacta aacctgacta cgacatctct
     2461 gccccttctt cgcggggcag tgcatgtaat cccttcagtt ggttggtaca acttgccaac
     2521 tgggccctgt tccacatgtg acacgggggg ggaccaaaca caaaggggtt ctctgactgt
     2581 agttgacatc cttataaatg gatgtgcaca tttgccaaca ctgagtggct ttcatcctgg
     2641 agcagacttt gcagtctgtg gactgcaaca caacattgcc tttatgtgta actcttggct
     2701 gaagctctta caccaatgct gggggacatg tacctcccag gggcccagga agactacggg
     2761 aggctacacc aacgtcaatc agaggggcct gtgtagctac cgataagcgg accctcaaga
     2821 gggcattagc aatagtgttt ataaggcccc cttgttaacc ctaaacgggt agcatatgct
     2881 tcccgggtag tagtatatac tatccagact aaccctaatt caatagcata tgttacccaa
     2941 cgggaagcat atgctatcga attagggtta gtaaaagggt cctaaggaac agcgatatct
     3001 cccaccccat gagctgtcac ggttttattt acatggggtc aggattccac gagggtagtg
     3061 aaccatttta gtcacaaggg cagtggctga agatcaagga gcgggcagtg aactctcctg
     3121 aatcttcgcc tgcttcttca ttctccttcg tttagctaat agaataactg ctgagttgtg
     3181 aacagtaagg tgtatgtgag gtgctcgaaa acaaggtttc aggtgacgcc cccagaataa
     3241 aatttggacg gggggttcag tggtggcatt gtgctatgac accaatataa ccctcacaaa
     3301 ccccttgggc aataaatact agtgtaggaa tgaaacattc tgaatatctt taacaataga
     3361 aatccatggg gtggggacaa gccgtaaaga ctggatgtcc atctcacacg aatttatggc
     3421 tatgggcaac acataatcct agtgcaatat gatactgggg ttattaagat gtgtcccagg
     3481 cagggaccaa gacaggtgaa ccatgttgtt acactctatt tgtaacaagg ggaaagagag
     3541 tggacgccga cagcagcgga ctccactggt tgtctctaac acccccgaaa attaaacggg
     3601 gctccacgcc aatggggccc ataaacaaag acaagtggcc actctttttt ttgaaattgt
     3661 ggagtggggg cacgcgtcag cccccacacg ccgccctgcg gttttggact gtaaaataag
     3721 ggtgtaataa cttggctgat tgtaaccccg ctaaccactg cggtcaaacc acttgcccac
     3781 aaaaccacta atggcacccc ggggaatacc tgcataagta ggtgggcggg ccaagatagg
     3841 ggcgcgattg ctgcgatctg gaggacaaat tacacacact tgcgcctgag cgccaagcac
     3901 agggttgttg gtcctcatat tcacgaggtc gctgagagca cggtgggcta atgttgccat
     3961 gggtagcata tactacccaa atatctggat agcatatgct atcctaatct atatctgggt
     4021 agcataggct atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt
     4081 agtatatgct atcctaattt atatctgggt agcataggct atcctaatct atatctgggt
     4141 agcatatgct atcctaatct atatctgggt agtatatgct atcctaatct gtatccgggt
     4201 agcatatgct atcctaatag agattagggt agtatatgct atcctaattt atatctgggt
     4261 agcatatact acccaaatat ctggatagca tatgctatcc taatctatat ctgggtagca
     4321 tatgctatcc taatctatat ctgggtagca taggctatcc taatctatat ctgggtagca
     4381 tatgctatcc taatctatat ctgggtagta tatgctatcc taatttatat ctgggtagca
     4441 taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagta
     4501 tatgctatcc taatctgtat ccgggtagca tatgctatcc tcatgataag ctgtcaaaca
     4561 tgagaatttt cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     4621 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     4681 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4741 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     4801 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     4861 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     4921 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     4981 acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa
     5041 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     5101 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     5161 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     5221 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     5281 gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg
     5341 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     5401 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     5461 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     5521 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     5581 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     5641 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     5701 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5761 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     5821 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     5881 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     5941 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     6001 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     6061 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     6121 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     6181 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     6241 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     6301 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     6361 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     6421 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     6481 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     6541 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     6601 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     6661 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     6721 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     6781 acaggaaaca gctatgacca tgattacgcc aagctctagc tagaggtcga ccaattctca
     6841 tgtttgacag cttatcatcg cagatccggg caacgttgtt gccattgctg caggcgcaga
     6901 actggtaggt atggaagatc tatacattga atcaatattg gcaattagcc atattagtca
     6961 ttggttatat agcataaatc aatattggct attggccatt gcatacgttg tatctatatc
     7021 ataatatgta catttatatt ggctcatgtc caatatgacc gc
//
