LOCUS       sequence-275111-        7689 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 7689)
  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 7689)
  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: D63_Ld_100680
FEATURES             Location/Qualifiers
     source          1..7689
                     /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             2853..2870
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2922..2977
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2976..2995)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5213..5231)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5299..5403
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5404..5472,5473..6264)
                     /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(5622..5641)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6435..7023
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6924..6943
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7177..7194
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7311..7332
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7347..7352,7353..7370,7371..7377)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7385..7401
                     /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     7390..7412
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7409..7425
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7409..7425
                     /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 gcggccgccc
     1201 agggcagtag cagtagcgac tactacgatg gaacatatta ctaccacgta atcttccgct
     1261 cggcaccagg cggcggagac tcgacagacg gaccctcagg agtaggcatt tatggcgtag
     1321 tcggatacgg cggagcaatg gtaggcggcg gtgtcgtatt cagcgatatg ggcggcagtg
     1381 gctccgcaat aaggctgata agtaaagcgg gctacctctc gaccatcgcc ggcaacaaca
     1441 ggaaaagggg agaccaagac gggcccagtc ccgatgcatt attcggtggc ataccaacag
     1501 gagacagcag agcaaactca cttgcctacg gcaacggtgg cttctactta gcggacacgc
     1561 tgaacaacgc cctgcgattc actaacgcta ccactaacgt gacaacaacc atactcaacg
     1621 cgaacacact caacacgcca aacagcctcg ccgtcagtat catctcaggg aaaaccagtg
     1681 ttttcatctc caacaccggc taccactccg tgctgtacat accaagcctg ggcagcccag
     1741 tgggccaagt gaacttcacc agtgacccat acttcgtccc cggcgcaata acagtactga
     1801 gccacctcag cagggccttc atagtgaacg ccacactcca aatgtactgc tggagctaca
     1861 tccaaccaag ctccaaatcc tggaaagtga gcaccccact ggcagcagac ttcggtagaa
     1921 tactgcaagc ctcaaaagat ggcacacaag tcctgtacgt gacggccaac ggctccacca
     1981 tagccggcct ggacgccaca gcatcatccg caacatcacc gtccttggtg ccacaaaaga
     2041 tgatagacct ggacacatcc cttatcggcg gaaagatcca actgttcttc gaacgaacat
     2101 ccgacagttg gtacatactt acaaccaccc aattcctcat cgtttccgca accccaattc
     2161 atccggaaga atcaagcagc tcaggctcct catcctcaag tagcagttca tccagtagca
     2221 gctcatcaag cagttcgtcc ggatcatcat cctcatcttc ctcgtcagaa ggcatcctgt
     2281 cagggcggca taaaggcatc gccgcattcc cgacggcagc attccccacc aacgacagtt
     2341 gcctaatgac gcaagtgtac aactggatga gaatcgacgc caccgaagcc tacaacacgg
     2401 acgacttcct gaacgaattc gtggtacccc ctacgaacat cacaacactg gtcgatggag
     2461 acgtcaacgt gagcacctgg tgcggcaaca taacgaccga ccgatcgaac gacggtacga
     2521 ttagcatcct gatcttctgg ggaccgagag gccacgacct gctttataca caagacaggc
     2581 tcgtgaaaag tccatggaaa cgcactcgcg ccttcctcaa cagtttaaac tccagcggct
     2641 ggaacctggg ccccttctgc ttctacaact gcacgacagc ctgcaaaacc atcacggcac
     2701 ccaaatgccc agcatacaac atcgggagcg catgcgacga cgtctgcaaa ggcgcaggcg
     2761 cgcccggcag cctgcaccac atcctggacg cccagaagat gctgtggaac cacagggaca
     2821 ggaacctgcc tcctctggcc cctctgggcc cccaccacca tcaccaccat tgagtctaga
     2881 cacgtgatta attaaggatc ccccgacctc gacctctggc taataaagga aatttatttt
     2941 cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc
     3001 aaatcatttg gtcgagatcc ctcggagatc tctagctaga ggatcgatcc ccgccccgga
     3061 cgaactaaac ctgactacga catctctgcc ccttcttcgc ggggcagtgc atgtaatccc
     3121 ttcagttggt tggtacaact tgccaactgg gccctgttcc acatgtgaca cgggggggga
     3181 ccaaacacaa aggggttctc tgactgtagt tgacatcctt ataaatggat gtgcacattt
     3241 gccaacactg agtggctttc atcctggagc agactttgca gtctgtggac tgcaacacaa
     3301 cattgccttt atgtgtaact cttggctgaa gctcttacac caatgctggg ggacatgtac
     3361 ctcccagggg cccaggaaga ctacgggagg ctacaccaac gtcaatcaga ggggcctgtg
     3421 tagctaccga taagcggacc ctcaagaggg cattagcaat agtgtttata aggccccctt
     3481 gttaacccta aacgggtagc atatgcttcc cgggtagtag tatatactat ccagactaac
     3541 cctaattcaa tagcatatgt tacccaacgg gaagcatatg ctatcgaatt agggttagta
     3601 aaagggtcct aaggaacagc gatatctccc accccatgag ctgtcacggt tttatttaca
     3661 tggggtcagg attccacgag ggtagtgaac cattttagtc acaagggcag tggctgaaga
     3721 tcaaggagcg ggcagtgaac tctcctgaat cttcgcctgc ttcttcattc tccttcgttt
     3781 agctaataga ataactgctg agttgtgaac agtaaggtgt atgtgaggtg ctcgaaaaca
     3841 aggtttcagg tgacgccccc agaataaaat ttggacgggg ggttcagtgg tggcattgtg
     3901 ctatgacacc aatataaccc tcacaaaccc cttgggcaat aaatactagt gtaggaatga
     3961 aacattctga atatctttaa caatagaaat ccatggggtg gggacaagcc gtaaagactg
     4021 gatgtccatc tcacacgaat ttatggctat gggcaacaca taatcctagt gcaatatgat
     4081 actggggtta ttaagatgtg tcccaggcag ggaccaagac aggtgaacca tgttgttaca
     4141 ctctatttgt aacaagggga aagagagtgg acgccgacag cagcggactc cactggttgt
     4201 ctctaacacc cccgaaaatt aaacggggct ccacgccaat ggggcccata aacaaagaca
     4261 agtggccact cttttttttg aaattgtgga gtgggggcac gcgtcagccc ccacacgccg
     4321 ccctgcggtt ttggactgta aaataagggt gtaataactt ggctgattgt aaccccgcta
     4381 accactgcgg tcaaaccact tgcccacaaa accactaatg gcaccccggg gaatacctgc
     4441 ataagtaggt gggcgggcca agataggggc gcgattgctg cgatctggag gacaaattac
     4501 acacacttgc gcctgagcgc caagcacagg gttgttggtc ctcatattca cgaggtcgct
     4561 gagagcacgg tgggctaatg ttgccatggg tagcatatac tacccaaata tctggatagc
     4621 atatgctatc ctaatctata tctgggtagc ataggctatc ctaatctata tctgggtagc
     4681 atatgctatc ctaatctata tctgggtagt atatgctatc ctaatttata tctgggtagc
     4741 ataggctatc ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt
     4801 atatgctatc ctaatctgta tccgggtagc atatgctatc ctaatagaga ttagggtagt
     4861 atatgctatc ctaatttata tctgggtagc atatactacc caaatatctg gatagcatat
     4921 gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagcatag
     4981 gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagtatat
     5041 gctatcctaa tttatatctg ggtagcatag gctatcctaa tctatatctg ggtagcatat
     5101 gctatcctaa tctatatctg ggtagtatat gctatcctaa tctgtatccg ggtagcatat
     5161 gctatcctca tgataagctg tcaaacatga gaattttctt gaagacgaaa gggcctcgtg
     5221 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     5281 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     5341 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     5401 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     5461 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     5521 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     5581 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     5641 tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     5701 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     5761 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     5821 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     5881 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     5941 atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     6001 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     6061 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     6121 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     6181 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     6241 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     6301 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     6361 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     6421 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6481 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6541 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     6601 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6661 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6721 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6781 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6841 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6901 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6961 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     7021 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     7081 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     7141 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     7201 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     7261 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     7321 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     7381 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     7441 ctctagctag aggtcgacca attctcatgt ttgacagctt atcatcgcag atccgggcaa
     7501 cgttgttgcc attgctgcag gcgcagaact ggtaggtatg gaagatctat acattgaatc
     7561 aatattggca attagccata ttagtcattg gttatatagc ataaatcaat attggctatt
     7621 ggccattgca tacgttgtat ctatatcata atatgtacat ttatattggc tcatgtccaa
     7681 tatgaccgc
//
