LOCUS       sequence-262354-        7569 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 7569)
  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 7569)
  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: D62_Ld_362720
FEATURES             Location/Qualifiers
     source          1..7569
                     /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             2733..2750
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2802..2857
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2856..2875)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5093..5111)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5179..5283
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5284..5352,5353..6144)
                     /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(5502..5521)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6315..6903
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6804..6823
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7057..7074
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7191..7212
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7227..7232,7233..7250,7251..7257)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7265..7281
                     /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     7270..7292
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7289..7305
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7289..7305
                     /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 agagcacaat cgccagagat ggcgccgctg acgccgccga catcctgaag gtgctgcagg
     1261 tcaacgtgct gcaccggcac ggcgccagaa gcggcctgcc tagagagaac accaccgaga
     1321 tctgcaccga gagcccctgc ggctacctga gctgggccgg catcgagatg ctgctgaaag
     1381 tgggcggctt cctgcggacc cggtacaaca ccgaccccag cgtggtgtcc agccccatgt
     1441 tcgagagccc caactacgac ctggacgtgg cctacagccg gtccaccgac gtgctgagaa
     1501 ccctgcagag cgccgaggcc ttcctgcggg gattcttccc caacatgtcc tccctgtacg
     1561 ccgccatcca caccatgccc gagagcaccg atgtgctgct gaacagcaac acccagccct
     1621 ggctgaagtt cttctacagc aacaacaagg ccctgctgcg ggccgtgtgc aaccccctga
     1681 ccgacgagct gttccccgac tggaccgaga tcaccaagat cggcgccgag atctaccagg
     1741 aaggctactg cagcgactac gagacacgca gcgactgcgt gcggaccctg ttcgacatcg
     1801 gagccgccaa gaaggccatc ggcgagctga accagtaccc cctgctggaa gccaacttcg
     1861 agaagctgaa gcgcgtgacc accgtgctgt tcgactacga gtaccactac aaccacagcg
     1921 accccctgat gttcaagcag ggcggcagag gccagccctt cgtgcagcag atggtcaaga
     1981 acatggaagg cgtgatcgag ggcagcaaca cctacaagct gatgcactac agcgcccacg
     2041 acaccagcct gggccctgtg tggggcaccc tgggcgatag aacccccgac ggcatgatgc
     2101 ccccattcgc ccaggtgctg gtggccgagc tgctgcagaa tcccagcaca ggcgcccact
     2161 acgtgcggat cctgagaggc aaccccggcc agagccccga caccaagttc gagttcggct
     2221 gggacagcaa gtggcagatg cagtgcatcg acgccctggg caccgcctac aaggccgagg
     2281 gcaacatctg ccccttcgcc gacttcaaga gattcgtgaa gtggtccgag cccgccgacg
     2341 ccagaggcta ctgctacctg gaccaggtgt ccatcgatat cgccgactgc cccctggaac
     2401 ccgtggccta cggaacccct cctaccgccc tggtgcccag cacctgtcgg ttctacagat
     2461 ccgcctgccc tcagttcgcc tgcggcatcg gcgagacact gaacagcgtg tccatgcagt
     2521 gcgtgtgcag caaggacagc tgcctgggcg gaaaggccaa cgaggaaagc tctggcgacg
     2581 ccgccgtgga cgccgacaag aagcccaccc tgagagacag caacggcctg agcggaggcg
     2641 cgcccggcag cctgcaccac atcctggacg cccagaagat gctgtggaac cacagggaca
     2701 ggaacctgcc tcctctggcc cctctgggcc cccaccacca tcaccaccat tgagtctaga
     2761 cacgtgatta attaaggatc ccccgacctc gacctctggc taataaagga aatttatttt
     2821 cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc
     2881 aaatcatttg gtcgagatcc ctcggagatc tctagctaga ggatcgatcc ccgccccgga
     2941 cgaactaaac ctgactacga catctctgcc ccttcttcgc ggggcagtgc atgtaatccc
     3001 ttcagttggt tggtacaact tgccaactgg gccctgttcc acatgtgaca cgggggggga
     3061 ccaaacacaa aggggttctc tgactgtagt tgacatcctt ataaatggat gtgcacattt
     3121 gccaacactg agtggctttc atcctggagc agactttgca gtctgtggac tgcaacacaa
     3181 cattgccttt atgtgtaact cttggctgaa gctcttacac caatgctggg ggacatgtac
     3241 ctcccagggg cccaggaaga ctacgggagg ctacaccaac gtcaatcaga ggggcctgtg
     3301 tagctaccga taagcggacc ctcaagaggg cattagcaat agtgtttata aggccccctt
     3361 gttaacccta aacgggtagc atatgcttcc cgggtagtag tatatactat ccagactaac
     3421 cctaattcaa tagcatatgt tacccaacgg gaagcatatg ctatcgaatt agggttagta
     3481 aaagggtcct aaggaacagc gatatctccc accccatgag ctgtcacggt tttatttaca
     3541 tggggtcagg attccacgag ggtagtgaac cattttagtc acaagggcag tggctgaaga
     3601 tcaaggagcg ggcagtgaac tctcctgaat cttcgcctgc ttcttcattc tccttcgttt
     3661 agctaataga ataactgctg agttgtgaac agtaaggtgt atgtgaggtg ctcgaaaaca
     3721 aggtttcagg tgacgccccc agaataaaat ttggacgggg ggttcagtgg tggcattgtg
     3781 ctatgacacc aatataaccc tcacaaaccc cttgggcaat aaatactagt gtaggaatga
     3841 aacattctga atatctttaa caatagaaat ccatggggtg gggacaagcc gtaaagactg
     3901 gatgtccatc tcacacgaat ttatggctat gggcaacaca taatcctagt gcaatatgat
     3961 actggggtta ttaagatgtg tcccaggcag ggaccaagac aggtgaacca tgttgttaca
     4021 ctctatttgt aacaagggga aagagagtgg acgccgacag cagcggactc cactggttgt
     4081 ctctaacacc cccgaaaatt aaacggggct ccacgccaat ggggcccata aacaaagaca
     4141 agtggccact cttttttttg aaattgtgga gtgggggcac gcgtcagccc ccacacgccg
     4201 ccctgcggtt ttggactgta aaataagggt gtaataactt ggctgattgt aaccccgcta
     4261 accactgcgg tcaaaccact tgcccacaaa accactaatg gcaccccggg gaatacctgc
     4321 ataagtaggt gggcgggcca agataggggc gcgattgctg cgatctggag gacaaattac
     4381 acacacttgc gcctgagcgc caagcacagg gttgttggtc ctcatattca cgaggtcgct
     4441 gagagcacgg tgggctaatg ttgccatggg tagcatatac tacccaaata tctggatagc
     4501 atatgctatc ctaatctata tctgggtagc ataggctatc ctaatctata tctgggtagc
     4561 atatgctatc ctaatctata tctgggtagt atatgctatc ctaatttata tctgggtagc
     4621 ataggctatc ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt
     4681 atatgctatc ctaatctgta tccgggtagc atatgctatc ctaatagaga ttagggtagt
     4741 atatgctatc ctaatttata tctgggtagc atatactacc caaatatctg gatagcatat
     4801 gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagcatag
     4861 gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagtatat
     4921 gctatcctaa tttatatctg ggtagcatag gctatcctaa tctatatctg ggtagcatat
     4981 gctatcctaa tctatatctg ggtagtatat gctatcctaa tctgtatccg ggtagcatat
     5041 gctatcctca tgataagctg tcaaacatga gaattttctt gaagacgaaa gggcctcgtg
     5101 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     5161 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     5221 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     5281 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     5341 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     5401 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     5461 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     5521 tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     5581 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     5641 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     5701 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     5761 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     5821 atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     5881 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     5941 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     6001 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     6061 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     6121 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     6181 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     6241 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     6301 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6361 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6421 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     6481 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6541 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6601 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6661 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6721 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6781 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6841 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6901 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     6961 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     7021 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     7081 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     7141 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     7201 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     7261 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     7321 ctctagctag aggtcgacca attctcatgt ttgacagctt atcatcgcag atccgggcaa
     7381 cgttgttgcc attgctgcag gcgcagaact ggtaggtatg gaagatctat acattgaatc
     7441 aatattggca attagccata ttagtcattg gttatatagc ataaatcaat attggctatt
     7501 ggccattgca tacgttgtat ctatatcata atatgtacat ttatattggc tcatgtccaa
     7561 tatgaccgc
//
