LOCUS       sequence-262359-        6879 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 6879)
  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 6879)
  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: D67_Ld_111030
FEATURES             Location/Qualifiers
     source          1..6879
                     /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             2043..2060
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2112..2167
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2166..2185)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4403..4421)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4489..4593
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4594..4662,4663..5454)
                     /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(4812..4831)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5625..6213
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6114..6133
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6367..6384
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6501..6522
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6537..6542,6543..6560,6561..6567)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6575..6591
                     /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     6580..6602
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6599..6615
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6599..6615
                     /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 caagcgcaat ggtcagtaaa gcaacaggcg gagcagttaa agcgatgcag gctaccagtg
     1261 ctgcgtccaa tgctatcgtc gttggtatgc aagccccgtt gggcgtgtat gcttgcgtgg
     1321 ctgaaccgcc gagcacagcc tctgggccct atgctggggt taaaacagtc gttgttcgag
     1381 ccattttgcc tcggcgggcc ccggatacta tgcaggtgcg agatatcctg gacgtatatc
     1441 ccgcctctgg catcacctgc gagaaggaaa cggcagaagc tgtggagaac tttaacaggg
     1501 cagccaaagt cgctgtagag aaagctaaag ctatgaaagc tactagggtg accctggtga
     1561 tgaagcccgc tacgaagtat gagagactga atggcttgtt tcgggaaacc tgcacgaaaa
     1621 ctatcgaggc tgcagggctc tcagtggaga cttccacaac agctgcagca accaatactc
     1681 tcattatgtt ccctgaaaag atgtctgtag tgatggtgag tgatgatccc gtatgtgaga
     1741 acgttcagta tgcgttcgcc ggggtcgtag gcggagtgca caccacttac tataccgatg
     1801 ctgggtctaa gattcatggt ggacacagtt ataaatccgt agctatggcc ctcgcagagg
     1861 aactcagatc tctgggcatg aaggctgaag cagccaaagt ggaagccgcc gctcagaagt
     1921 ctcccaggaa cgcagctgcg gctatgggcg cgcccggcag cctgcaccac atcctggacg
     1981 cccagaagat gctgtggaac cacagggaca ggaacctgcc tcctctggcc cctctgggcc
     2041 cccaccacca tcaccaccat tgagtctaga cacgtgatta attaaggatc ccccgacctc
     2101 gacctctggc taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg
     2161 tctctcactc ggaaggacat atgggagggc aaatcatttg gtcgagatcc ctcggagatc
     2221 tctagctaga ggatcgatcc ccgccccgga cgaactaaac ctgactacga catctctgcc
     2281 ccttcttcgc ggggcagtgc atgtaatccc ttcagttggt tggtacaact tgccaactgg
     2341 gccctgttcc acatgtgaca cgggggggga ccaaacacaa aggggttctc tgactgtagt
     2401 tgacatcctt ataaatggat gtgcacattt gccaacactg agtggctttc atcctggagc
     2461 agactttgca gtctgtggac tgcaacacaa cattgccttt atgtgtaact cttggctgaa
     2521 gctcttacac caatgctggg ggacatgtac ctcccagggg cccaggaaga ctacgggagg
     2581 ctacaccaac gtcaatcaga ggggcctgtg tagctaccga taagcggacc ctcaagaggg
     2641 cattagcaat agtgtttata aggccccctt gttaacccta aacgggtagc atatgcttcc
     2701 cgggtagtag tatatactat ccagactaac cctaattcaa tagcatatgt tacccaacgg
     2761 gaagcatatg ctatcgaatt agggttagta aaagggtcct aaggaacagc gatatctccc
     2821 accccatgag ctgtcacggt tttatttaca tggggtcagg attccacgag ggtagtgaac
     2881 cattttagtc acaagggcag tggctgaaga tcaaggagcg ggcagtgaac tctcctgaat
     2941 cttcgcctgc ttcttcattc tccttcgttt agctaataga ataactgctg agttgtgaac
     3001 agtaaggtgt atgtgaggtg ctcgaaaaca aggtttcagg tgacgccccc agaataaaat
     3061 ttggacgggg ggttcagtgg tggcattgtg ctatgacacc aatataaccc tcacaaaccc
     3121 cttgggcaat aaatactagt gtaggaatga aacattctga atatctttaa caatagaaat
     3181 ccatggggtg gggacaagcc gtaaagactg gatgtccatc tcacacgaat ttatggctat
     3241 gggcaacaca taatcctagt gcaatatgat actggggtta ttaagatgtg tcccaggcag
     3301 ggaccaagac aggtgaacca tgttgttaca ctctatttgt aacaagggga aagagagtgg
     3361 acgccgacag cagcggactc cactggttgt ctctaacacc cccgaaaatt aaacggggct
     3421 ccacgccaat ggggcccata aacaaagaca agtggccact cttttttttg aaattgtgga
     3481 gtgggggcac gcgtcagccc ccacacgccg ccctgcggtt ttggactgta aaataagggt
     3541 gtaataactt ggctgattgt aaccccgcta accactgcgg tcaaaccact tgcccacaaa
     3601 accactaatg gcaccccggg gaatacctgc ataagtaggt gggcgggcca agataggggc
     3661 gcgattgctg cgatctggag gacaaattac acacacttgc gcctgagcgc caagcacagg
     3721 gttgttggtc ctcatattca cgaggtcgct gagagcacgg tgggctaatg ttgccatggg
     3781 tagcatatac tacccaaata tctggatagc atatgctatc ctaatctata tctgggtagc
     3841 ataggctatc ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt
     3901 atatgctatc ctaatttata tctgggtagc ataggctatc ctaatctata tctgggtagc
     3961 atatgctatc ctaatctata tctgggtagt atatgctatc ctaatctgta tccgggtagc
     4021 atatgctatc ctaatagaga ttagggtagt atatgctatc ctaatttata tctgggtagc
     4081 atatactacc caaatatctg gatagcatat gctatcctaa tctatatctg ggtagcatat
     4141 gctatcctaa tctatatctg ggtagcatag gctatcctaa tctatatctg ggtagcatat
     4201 gctatcctaa tctatatctg ggtagtatat gctatcctaa tttatatctg ggtagcatag
     4261 gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagtatat
     4321 gctatcctaa tctgtatccg ggtagcatat gctatcctca tgataagctg tcaaacatga
     4381 gaattttctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     4441 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     4501 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4561 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     4621 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     4681 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     4741 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     4801 tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc
     4861 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     4921 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     4981 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     5041 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     5101 gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac aacgttgcgc
     5161 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     5221 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     5281 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     5341 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     5401 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     5461 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5521 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     5581 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5641 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5701 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     5761 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     5821 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     5881 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     5941 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6001 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6061 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6121 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6181 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6241 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     6301 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     6361 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     6421 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     6481 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     6541 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     6601 ggaaacagct atgaccatga ttacgccaag ctctagctag aggtcgacca attctcatgt
     6661 ttgacagctt atcatcgcag atccgggcaa cgttgttgcc attgctgcag gcgcagaact
     6721 ggtaggtatg gaagatctat acattgaatc aatattggca attagccata ttagtcattg
     6781 gttatatagc ataaatcaat attggctatt ggccattgca tacgttgtat ctatatcata
     6841 atatgtacat ttatattggc tcatgtccaa tatgaccgc
//
