LOCUS       sequence-262340-        7779 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 7779)
  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 7779)
  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: D48_Ld_353820
FEATURES             Location/Qualifiers
     source          1..7779
                     /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             2943..2960
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3012..3067
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3066..3085)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5303..5321)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5389..5493
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5494..5562,5563..6354)
                     /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(5712..5731)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6525..7113
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7014..7033
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7267..7284
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7401..7422
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7437..7442,7443..7460,7461..7467)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7475..7491
                     /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     7480..7502
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7499..7515
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7499..7515
                     /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 ccgtgaacct gctgcctctg atgggcgagg gcctgttcga cgtggaagtc gtgtgccgca
     1261 gcgccatgcc cagcgagccc accgtgtacg acagaagcgt gatcagcgcc ttccggttca
     1321 gcgtgaacgc cagcgccgac agcaccaaag aaaaagtgca tctgcccctg tacgtggaac
     1381 ggatcctgga cagcgactac gtgcggacct actacatcaa cagcaccgac aagtacagca
     1441 tcagcgacag caagggctgc gtgcacacca agggactgta cgagagcagc gacgaggtgc
     1501 tgctgagcga gagcctgatc accatgctgc ggagccccct gagcgccacc cccagcccta
     1561 gcatgatcag aggaatcgcc gccgacaact actccagcca gtacaacctg accatcccca
     1621 gcagctacgg cggcctgggc agccctgcca catatagagc cgccgtgctg acctccaccc
     1681 agcactggct ggtcaacctg atgcaggtcg acctcacccc cgtgtccatc gaagtgatga
     1741 ccaccgacag cgcctccaag atcaacaact gcctgttcac cttcagcttt ttcggcagca
     1801 atgtgtctgc cgtgcggggc tccctgccca gctcctgcaa gaacagcagc tccgagcacc
     1861 ccagagtgtg gaaaagcagc aacgtggccc tgcctatgat gcgggccctg gctggcgccg
     1921 ctaaggcctt cagcagcaga cccagcgtgt tcagctccag cagcgagaca agctcctcca
     1981 ccagcgctgc cgacaccgcc aactacacca tgcccgcctt ccccgacgac tttaccgcca
     2041 atttcctggt ggtgtccccc ctgaagaagt ccttctacca ggtccgagag agctacagcc
     2101 tgtacgccgg cttcagcctg accagcctgc agttccctct gaagggcatt gccggcagag
     2161 tgtacaagca cgagtggtac accaacagct ggaaccagat gagctactac cacaccgagt
     2221 tcgccgtgcc ctccggccag gaaatggccg acgaggccct gcggaagtac ttcttccctg
     2281 acctgaacac ctgtcggcgg gtcgtgatcg cctacgacct gatggcccgg tctagcagct
     2341 ctctgctgct gtacagcccc aacgtgcccc ccaccttcat cggcaaccag accgtgcgga
     2401 acatcccctg cggcgtgtgg gtggccgaga tccagggcgt cagagtgacc tggtactggg
     2461 ccaccaccga tctggtggat accgccagct tctggaccgt ggacagcatc gagagcggcg
     2521 cctccgccta caccaggctc gtgcggatga cagtgaccgg caaaggcggc gctccccccc
     2581 tgtttgccca ccacccattc tttccacagt cctacgcctt tcccgtggac gaccgggatg
     2641 tggcctgcac cgccatgatg cctagcgagg ccgacatggg ctgctacagc tacgccaaga
     2701 acgccgatta cacctacatc tacgagatca ccagcttcgt gccctatgtg cggcggaacg
     2761 actaccggct gcctgccgcc tgtgtgggcg tgaagatcct gggctccatc cctagcttcc
     2821 agtgcaacta caagggcatc accggaggcg cgcccggcag cctgcaccac atcctggacg
     2881 cccagaagat gctgtggaac cacagggaca ggaacctgcc tcctctggcc cctctgggcc
     2941 cccaccacca tcaccaccat tgagtctaga cacgtgatta attaaggatc ccccgacctc
     3001 gacctctggc taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg
     3061 tctctcactc ggaaggacat atgggagggc aaatcatttg gtcgagatcc ctcggagatc
     3121 tctagctaga ggatcgatcc ccgccccgga cgaactaaac ctgactacga catctctgcc
     3181 ccttcttcgc ggggcagtgc atgtaatccc ttcagttggt tggtacaact tgccaactgg
     3241 gccctgttcc acatgtgaca cgggggggga ccaaacacaa aggggttctc tgactgtagt
     3301 tgacatcctt ataaatggat gtgcacattt gccaacactg agtggctttc atcctggagc
     3361 agactttgca gtctgtggac tgcaacacaa cattgccttt atgtgtaact cttggctgaa
     3421 gctcttacac caatgctggg ggacatgtac ctcccagggg cccaggaaga ctacgggagg
     3481 ctacaccaac gtcaatcaga ggggcctgtg tagctaccga taagcggacc ctcaagaggg
     3541 cattagcaat agtgtttata aggccccctt gttaacccta aacgggtagc atatgcttcc
     3601 cgggtagtag tatatactat ccagactaac cctaattcaa tagcatatgt tacccaacgg
     3661 gaagcatatg ctatcgaatt agggttagta aaagggtcct aaggaacagc gatatctccc
     3721 accccatgag ctgtcacggt tttatttaca tggggtcagg attccacgag ggtagtgaac
     3781 cattttagtc acaagggcag tggctgaaga tcaaggagcg ggcagtgaac tctcctgaat
     3841 cttcgcctgc ttcttcattc tccttcgttt agctaataga ataactgctg agttgtgaac
     3901 agtaaggtgt atgtgaggtg ctcgaaaaca aggtttcagg tgacgccccc agaataaaat
     3961 ttggacgggg ggttcagtgg tggcattgtg ctatgacacc aatataaccc tcacaaaccc
     4021 cttgggcaat aaatactagt gtaggaatga aacattctga atatctttaa caatagaaat
     4081 ccatggggtg gggacaagcc gtaaagactg gatgtccatc tcacacgaat ttatggctat
     4141 gggcaacaca taatcctagt gcaatatgat actggggtta ttaagatgtg tcccaggcag
     4201 ggaccaagac aggtgaacca tgttgttaca ctctatttgt aacaagggga aagagagtgg
     4261 acgccgacag cagcggactc cactggttgt ctctaacacc cccgaaaatt aaacggggct
     4321 ccacgccaat ggggcccata aacaaagaca agtggccact cttttttttg aaattgtgga
     4381 gtgggggcac gcgtcagccc ccacacgccg ccctgcggtt ttggactgta aaataagggt
     4441 gtaataactt ggctgattgt aaccccgcta accactgcgg tcaaaccact tgcccacaaa
     4501 accactaatg gcaccccggg gaatacctgc ataagtaggt gggcgggcca agataggggc
     4561 gcgattgctg cgatctggag gacaaattac acacacttgc gcctgagcgc caagcacagg
     4621 gttgttggtc ctcatattca cgaggtcgct gagagcacgg tgggctaatg ttgccatggg
     4681 tagcatatac tacccaaata tctggatagc atatgctatc ctaatctata tctgggtagc
     4741 ataggctatc ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt
     4801 atatgctatc ctaatttata tctgggtagc ataggctatc ctaatctata tctgggtagc
     4861 atatgctatc ctaatctata tctgggtagt atatgctatc ctaatctgta tccgggtagc
     4921 atatgctatc ctaatagaga ttagggtagt atatgctatc ctaatttata tctgggtagc
     4981 atatactacc caaatatctg gatagcatat gctatcctaa tctatatctg ggtagcatat
     5041 gctatcctaa tctatatctg ggtagcatag gctatcctaa tctatatctg ggtagcatat
     5101 gctatcctaa tctatatctg ggtagtatat gctatcctaa tttatatctg ggtagcatag
     5161 gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagtatat
     5221 gctatcctaa tctgtatccg ggtagcatat gctatcctca tgataagctg tcaaacatga
     5281 gaattttctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     5341 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     5401 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     5461 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     5521 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     5581 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     5641 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     5701 tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc
     5761 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     5821 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     5881 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     5941 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     6001 gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac aacgttgcgc
     6061 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     6121 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     6181 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     6241 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     6301 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     6361 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     6421 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     6481 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     6541 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     6601 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     6661 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     6721 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     6781 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     6841 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6901 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6961 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     7021 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     7081 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     7141 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     7201 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     7261 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     7321 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     7381 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     7441 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     7501 ggaaacagct atgaccatga ttacgccaag ctctagctag aggtcgacca attctcatgt
     7561 ttgacagctt atcatcgcag atccgggcaa cgttgttgcc attgctgcag gcgcagaact
     7621 ggtaggtatg gaagatctat acattgaatc aatattggca attagccata ttagtcattg
     7681 gttatatagc ataaatcaat attggctatt ggccattgca tacgttgtat ctatatcata
     7741 atatgtacat ttatattggc tcatgtccaa tatgaccgc
//
