LOCUS       sequence-262325-        7389 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 7389)
  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 7389)
  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: D30_Ld_260440
FEATURES             Location/Qualifiers
     source          1..7389
                     /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             2553..2570
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2622..2677
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2676..2695)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4913..4931)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4999..5103
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5104..5172,5173..5964)
                     /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(5322..5341)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6135..6723
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6624..6643
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6877..6894
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7011..7032
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7047..7052,7053..7070,7071..7077)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7085..7101
                     /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     7090..7112
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7109..7125
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7109..7125
                     /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 cccagaacaa tggctggcgg agcatgtgca tcggccggct ggcctacaag tgcagcagcg
     1261 agtggtgccg gtccagcgtg atcaccccca caagcgtgtc cctggccagc agctgcgtgg
     1321 gcaagggcaa cgccctggtg ctgatcgctc tggtggaagt gggccccagc gtggtgtcca
     1381 gacggggcga gaacatcagc aagtacatgg ccgacgagct gcggcggcag atcgacagcg
     1441 gcgagtttca gagagccgtg cggctgaccc tggccgctcc cagattcgcc agcgacaaca
     1501 ccggcgacga ccccagaacc agcctgaccg agctgtttag agagaaggcc agcctgcggg
     1561 ccatcttcga cgtgcccaga aaccagaccc ggtacatggt ggtcaacgac gtggtgcccc
     1621 tgcccgccgt ggacggcctg ggcaatttca gctacaccac ctgggaagag gccgtggatg
     1681 ccgccctgcc tgtgtccaag gtggacgacg cctcccagct gccccggtac ttccccaacc
     1741 tgtacaccaa gctggccgcc ttcagcaacg agagggccgc ctctgccgcc ccacctagca
     1801 gcaagcagtg cgacgcactg gtggtggtcg tgggcacaat cgcccaccct ctggcctctg
     1861 tgcggagcgt gtggcagacc cctagagatg ctggcgccca gctgagcctg ggccacgaat
     1921 atgtgcgggg agccgtgacc agatacctga tgacctgccg gaagttcctg cggaacgtgc
     1981 acggcgtgta ctacctgcag tacgagagcc ggctgcccgc ttgggccttt acagccgagg
     2041 tgctgggcgg ccagagagag gaactgagca tcatcaccgc caccaacgcc accgccgaca
     2101 gagccccagc cctgggaaac acctccagct tcgccgctgc cttctgcacc agcgccaatg
     2161 ccacagcccc tagcgcccct ctgaggcctt gcgagcaggc ctgcagaatc tacggccgga
     2221 tcgcctccga cctgatcgcc ggcgacagag gaatcgcccg gacctccttc gagaagcaca
     2281 accagttcag cagcagctcc ttcacctaca actgctacta cgaagtgctg tccgcccagg
     2341 gcgaggccgc catcaatggc accttcgacg ccaagaccgt gtcccgggtg ttcgacgact
     2401 gcctgcacct gagcccagcc ggcagcagag cctacgcccg gtgcctgctg agaagaggcg
     2461 cgcccggcag cctgcaccac atcctggacg cccagaagat gctgtggaac cacagggaca
     2521 ggaacctgcc tcctctggcc cctctgggcc cccaccacca tcaccaccat tgagtctaga
     2581 cacgtgatta attaaggatc ccccgacctc gacctctggc taataaagga aatttatttt
     2641 cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc
     2701 aaatcatttg gtcgagatcc ctcggagatc tctagctaga ggatcgatcc ccgccccgga
     2761 cgaactaaac ctgactacga catctctgcc ccttcttcgc ggggcagtgc atgtaatccc
     2821 ttcagttggt tggtacaact tgccaactgg gccctgttcc acatgtgaca cgggggggga
     2881 ccaaacacaa aggggttctc tgactgtagt tgacatcctt ataaatggat gtgcacattt
     2941 gccaacactg agtggctttc atcctggagc agactttgca gtctgtggac tgcaacacaa
     3001 cattgccttt atgtgtaact cttggctgaa gctcttacac caatgctggg ggacatgtac
     3061 ctcccagggg cccaggaaga ctacgggagg ctacaccaac gtcaatcaga ggggcctgtg
     3121 tagctaccga taagcggacc ctcaagaggg cattagcaat agtgtttata aggccccctt
     3181 gttaacccta aacgggtagc atatgcttcc cgggtagtag tatatactat ccagactaac
     3241 cctaattcaa tagcatatgt tacccaacgg gaagcatatg ctatcgaatt agggttagta
     3301 aaagggtcct aaggaacagc gatatctccc accccatgag ctgtcacggt tttatttaca
     3361 tggggtcagg attccacgag ggtagtgaac cattttagtc acaagggcag tggctgaaga
     3421 tcaaggagcg ggcagtgaac tctcctgaat cttcgcctgc ttcttcattc tccttcgttt
     3481 agctaataga ataactgctg agttgtgaac agtaaggtgt atgtgaggtg ctcgaaaaca
     3541 aggtttcagg tgacgccccc agaataaaat ttggacgggg ggttcagtgg tggcattgtg
     3601 ctatgacacc aatataaccc tcacaaaccc cttgggcaat aaatactagt gtaggaatga
     3661 aacattctga atatctttaa caatagaaat ccatggggtg gggacaagcc gtaaagactg
     3721 gatgtccatc tcacacgaat ttatggctat gggcaacaca taatcctagt gcaatatgat
     3781 actggggtta ttaagatgtg tcccaggcag ggaccaagac aggtgaacca tgttgttaca
     3841 ctctatttgt aacaagggga aagagagtgg acgccgacag cagcggactc cactggttgt
     3901 ctctaacacc cccgaaaatt aaacggggct ccacgccaat ggggcccata aacaaagaca
     3961 agtggccact cttttttttg aaattgtgga gtgggggcac gcgtcagccc ccacacgccg
     4021 ccctgcggtt ttggactgta aaataagggt gtaataactt ggctgattgt aaccccgcta
     4081 accactgcgg tcaaaccact tgcccacaaa accactaatg gcaccccggg gaatacctgc
     4141 ataagtaggt gggcgggcca agataggggc gcgattgctg cgatctggag gacaaattac
     4201 acacacttgc gcctgagcgc caagcacagg gttgttggtc ctcatattca cgaggtcgct
     4261 gagagcacgg tgggctaatg ttgccatggg tagcatatac tacccaaata tctggatagc
     4321 atatgctatc ctaatctata tctgggtagc ataggctatc ctaatctata tctgggtagc
     4381 atatgctatc ctaatctata tctgggtagt atatgctatc ctaatttata tctgggtagc
     4441 ataggctatc ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt
     4501 atatgctatc ctaatctgta tccgggtagc atatgctatc ctaatagaga ttagggtagt
     4561 atatgctatc ctaatttata tctgggtagc atatactacc caaatatctg gatagcatat
     4621 gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagcatag
     4681 gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagtatat
     4741 gctatcctaa tttatatctg ggtagcatag gctatcctaa tctatatctg ggtagcatat
     4801 gctatcctaa tctatatctg ggtagtatat gctatcctaa tctgtatccg ggtagcatat
     4861 gctatcctca tgataagctg tcaaacatga gaattttctt gaagacgaaa gggcctcgtg
     4921 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     4981 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     5041 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     5101 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     5161 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     5221 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     5281 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     5341 tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     5401 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     5461 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     5521 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     5581 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     5641 atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     5701 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     5761 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     5821 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     5881 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     5941 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     6001 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     6061 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     6121 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6181 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6241 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     6301 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6361 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6421 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6481 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6541 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6601 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6661 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6721 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     6781 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     6841 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     6901 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     6961 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     7021 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     7081 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     7141 ctctagctag aggtcgacca attctcatgt ttgacagctt atcatcgcag atccgggcaa
     7201 cgttgttgcc attgctgcag gcgcagaact ggtaggtatg gaagatctat acattgaatc
     7261 aatattggca attagccata ttagtcattg gttatatagc ataaatcaat attggctatt
     7321 ggccattgca tacgttgtat ctatatcata atatgtacat ttatattggc tcatgtccaa
     7381 tatgaccgc
//
