LOCUS       sequence-262339-        7479 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 7479)
  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 7479)
  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: D46_Ld_362700
FEATURES             Location/Qualifiers
     source          1..7479
                     /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             2643..2660
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2712..2767
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2766..2785)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5003..5021)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5089..5193
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5194..5262,5263..6054)
                     /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(5412..5431)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6225..6813
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6714..6733
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6967..6984
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7101..7122
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7137..7142,7143..7160,7161..7167)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7175..7191
                     /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     7180..7202
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7199..7215
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7199..7215
                     /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 gcggccgcca
     1201 ccgcccccat gtacaaggtg gaactggtgc aggtcgtgca ccggcacggc gccagatccc
     1261 ccctggtgga cgacaaccac accctgatct gcggcaccga gttcccttgc ggcttcctga
     1321 actacgaggg ccaggccatg ctggtcaagc tgggcgagta cctgagacac cggtacaccg
     1381 aggaccgcag cgtggtgtcc aagccctact tcccctacag ctggtacaac ctgagcatca
     1441 gctacacccg gtccaccgac gtgctgcgga ccctgcagtc tgccaacggc ctgctgagag
     1501 gcctgttccc caacaacagc aacttcttcc ccgccatcca caccgtgggc ttcgaggaag
     1561 atgtgctgct gcacagcatg atggtgccca tgatccgggc cagattcaac tacgccaaag
     1621 aggaactgcg ggccgtctgc gaccagatgc tggaccggct gatgcccttc gaccagctgc
     1681 aggccgtggc cgccgaagtg cacagccagg gcttttgcgc caactacaca ctgcggagca
     1741 gatgcgccga gcggctgtgc gacatcggca gagcctacga gcccaccggc agactggaac
     1801 ggctgcccct gctgtccaga cacctggacg atgtgtgcgc cgtgaccgcc atgagcagcc
     1861 acttttgctt cgcctacaac gccagcaacc ccgtgcacca gaagcagggc gctcccttct
     1921 accacctggc ccggctgctg gtgtctaaca tgctggccca ccagcagcgg gaaaccgccc
     1981 caccctacaa gctgtacgag tacagcgccc acgacaccac catcagcccc ctggccgtgt
     2041 ccttcggcga cagcagcctg gaagctatgc tgcccccctt cggcaccgcc ttcatcatcg
     2101 agctgctgag cctgaccgac gcccctgccg ccagcagcag attctacgtc agactgctgc
     2161 ggggccacag cggcgtgcgg cccgagagca atttcacctt cgccctgagc cacttcgaca
     2221 tgcggtgcaa ggacgccaca ggcaacacct atatcgccac cgacaacatc tgccccttca
     2281 ccgacttcga gcggttcatc aacagcacag cccctaccag ccccatgggc acctgttacc
     2341 tggaccccgg actgctgttc cggatggact gccctgtgga cgccgtgggc gacaacagaa
     2401 gcctgagcaa ggattgcctg ttctaccgga agcactgcag caattacagc tgtggcaccg
     2461 gctactacct ggatgccatc gactacggct gtcaccggat ccctgccaac aactccacca
     2521 ccgtgtccag cccaatgtcc agcggaggcg cgcccggcag cctgcaccac atcctggacg
     2581 cccagaagat gctgtggaac cacagggaca ggaacctgcc tcctctggcc cctctgggcc
     2641 cccaccacca tcaccaccat tgagtctaga cacgtgatta attaaggatc ccccgacctc
     2701 gacctctggc taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg
     2761 tctctcactc ggaaggacat atgggagggc aaatcatttg gtcgagatcc ctcggagatc
     2821 tctagctaga ggatcgatcc ccgccccgga cgaactaaac ctgactacga catctctgcc
     2881 ccttcttcgc ggggcagtgc atgtaatccc ttcagttggt tggtacaact tgccaactgg
     2941 gccctgttcc acatgtgaca cgggggggga ccaaacacaa aggggttctc tgactgtagt
     3001 tgacatcctt ataaatggat gtgcacattt gccaacactg agtggctttc atcctggagc
     3061 agactttgca gtctgtggac tgcaacacaa cattgccttt atgtgtaact cttggctgaa
     3121 gctcttacac caatgctggg ggacatgtac ctcccagggg cccaggaaga ctacgggagg
     3181 ctacaccaac gtcaatcaga ggggcctgtg tagctaccga taagcggacc ctcaagaggg
     3241 cattagcaat agtgtttata aggccccctt gttaacccta aacgggtagc atatgcttcc
     3301 cgggtagtag tatatactat ccagactaac cctaattcaa tagcatatgt tacccaacgg
     3361 gaagcatatg ctatcgaatt agggttagta aaagggtcct aaggaacagc gatatctccc
     3421 accccatgag ctgtcacggt tttatttaca tggggtcagg attccacgag ggtagtgaac
     3481 cattttagtc acaagggcag tggctgaaga tcaaggagcg ggcagtgaac tctcctgaat
     3541 cttcgcctgc ttcttcattc tccttcgttt agctaataga ataactgctg agttgtgaac
     3601 agtaaggtgt atgtgaggtg ctcgaaaaca aggtttcagg tgacgccccc agaataaaat
     3661 ttggacgggg ggttcagtgg tggcattgtg ctatgacacc aatataaccc tcacaaaccc
     3721 cttgggcaat aaatactagt gtaggaatga aacattctga atatctttaa caatagaaat
     3781 ccatggggtg gggacaagcc gtaaagactg gatgtccatc tcacacgaat ttatggctat
     3841 gggcaacaca taatcctagt gcaatatgat actggggtta ttaagatgtg tcccaggcag
     3901 ggaccaagac aggtgaacca tgttgttaca ctctatttgt aacaagggga aagagagtgg
     3961 acgccgacag cagcggactc cactggttgt ctctaacacc cccgaaaatt aaacggggct
     4021 ccacgccaat ggggcccata aacaaagaca agtggccact cttttttttg aaattgtgga
     4081 gtgggggcac gcgtcagccc ccacacgccg ccctgcggtt ttggactgta aaataagggt
     4141 gtaataactt ggctgattgt aaccccgcta accactgcgg tcaaaccact tgcccacaaa
     4201 accactaatg gcaccccggg gaatacctgc ataagtaggt gggcgggcca agataggggc
     4261 gcgattgctg cgatctggag gacaaattac acacacttgc gcctgagcgc caagcacagg
     4321 gttgttggtc ctcatattca cgaggtcgct gagagcacgg tgggctaatg ttgccatggg
     4381 tagcatatac tacccaaata tctggatagc atatgctatc ctaatctata tctgggtagc
     4441 ataggctatc ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt
     4501 atatgctatc ctaatttata tctgggtagc ataggctatc ctaatctata tctgggtagc
     4561 atatgctatc ctaatctata tctgggtagt atatgctatc ctaatctgta tccgggtagc
     4621 atatgctatc ctaatagaga ttagggtagt atatgctatc ctaatttata tctgggtagc
     4681 atatactacc caaatatctg gatagcatat gctatcctaa tctatatctg ggtagcatat
     4741 gctatcctaa tctatatctg ggtagcatag gctatcctaa tctatatctg ggtagcatat
     4801 gctatcctaa tctatatctg ggtagtatat gctatcctaa tttatatctg ggtagcatag
     4861 gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagtatat
     4921 gctatcctaa tctgtatccg ggtagcatat gctatcctca tgataagctg tcaaacatga
     4981 gaattttctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     5041 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     5101 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     5161 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     5221 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     5281 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     5341 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     5401 tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc
     5461 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     5521 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     5581 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     5641 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     5701 gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac aacgttgcgc
     5761 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     5821 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     5881 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     5941 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     6001 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     6061 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     6121 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     6181 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     6241 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     6301 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     6361 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     6421 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     6481 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     6541 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6601 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6661 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6721 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6781 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6841 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     6901 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     6961 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     7021 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     7081 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     7141 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     7201 ggaaacagct atgaccatga ttacgccaag ctctagctag aggtcgacca attctcatgt
     7261 ttgacagctt atcatcgcag atccgggcaa cgttgttgcc attgctgcag gcgcagaact
     7321 ggtaggtatg gaagatctat acattgaatc aatattggca attagccata ttagtcattg
     7381 gttatatagc ataaatcaat attggctatt ggccattgca tacgttgtat ctatatcata
     7441 atatgtacat ttatattggc tcatgtccaa tatgaccgc
//
