LOCUS       sequence-274677-        6573 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 6573)
  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 6573)
  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: D88_Ld_320450
FEATURES             Location/Qualifiers
     source          1..6573
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     1086..1105
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1128..1137
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1731..1748
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1800..1855
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1854..1873)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4091..4109)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4177..4281
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4282..4350,4351..5142)
                     /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(4500..4519)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5313..5901
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5802..5821
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6055..6072
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6189..6210
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6225..6230,6231..6248,6249..6255)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6263..6279
                     /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     6268..6290
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6287..6303
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6287..6303
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agatcctcac tctcttccgc atcgctgtct gcgagggcca gctgttgggc tcgcggttga
      661 ggacaaactc ttcgcggtct ttccagtact cttggatcgg aaacccgtcg gcctccgaac
      721 ggtactccgc caccgaggga cctgagcgag tccgcatcga ccggatcgga aaacctctcg
      781 agaaaggcgt ctaaccagtc acagtcgcaa ggtaggctga gcaccgtggc gggcggcagc
      841 gggtggcggt cggggttgtt tctggcggag gtgctgctga tgatgtaatt aaagtaggcg
      901 gtcttgagac ggcggatggt cgaggtgagg tgtggcaggc ttgagatcca gctgttgggg
      961 tgagtactcc ctctcaaaag cgggcattac ttctgcgcta agattgtcag tttccaaaaa
     1021 cgaggaggat ttgatattca cctggcccga tctggccata cacttgagtg acaatgacat
     1081 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caagtttgcc accatggagt
     1141 ttcagaccca ggtactcatg tccctgctgc tctgcatgtc tggtgcggcc gcctctacct
     1201 ggaactcaac tcctgaaaag tatggcgagt ctgcagctgc gagcggagcc acagtgttgc
     1261 cgggtgtgga cctcgctcgc agggccgtct ccccaaatca ggaatttcca gcgacctggt
     1321 ctgaagacga acctctgtca ccatccaacc ctagagtttg ctggcgcgaa cactactcac
     1381 tggatgctca aaagccgtat taccaaaata ttaagacaat ggaagtgaca tgggatattc
     1441 ccgaaggctt tgtaaccaga tttccacgac tttatgcaag caacggctac catgtcgacg
     1501 cgacaggagc agtgtttcgg tcctcaaccg ccagtgagcc acccgcaaat ggaaccgtcg
     1561 ggacaacagt cgcaaaagag gccggtgtga aagcagtcag tctgaaacag cgactggccg
     1621 cctatggagc tggcggcgcg cccggcagcc tgcaccacat cctggacgcc cagaagatgc
     1681 tgtggaacca cagggacagg aacctgcctc ctctggcccc tctgggcccc caccaccatc
     1741 accaccattg agtctagaca cgtgattaat taaggatccc ccgacctcga cctctggcta
     1801 ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg
     1861 aaggacatat gggagggcaa atcatttggt cgagatccct cggagatctc tagctagagg
     1921 atcgatcccc gccccggacg aactaaacct gactacgaca tctctgcccc ttcttcgcgg
     1981 ggcagtgcat gtaatccctt cagttggttg gtacaacttg ccaactgggc cctgttccac
     2041 atgtgacacg gggggggacc aaacacaaag gggttctctg actgtagttg acatccttat
     2101 aaatggatgt gcacatttgc caacactgag tggctttcat cctggagcag actttgcagt
     2161 ctgtggactg caacacaaca ttgcctttat gtgtaactct tggctgaagc tcttacacca
     2221 atgctggggg acatgtacct cccaggggcc caggaagact acgggaggct acaccaacgt
     2281 caatcagagg ggcctgtgta gctaccgata agcggaccct caagagggca ttagcaatag
     2341 tgtttataag gcccccttgt taaccctaaa cgggtagcat atgcttcccg ggtagtagta
     2401 tatactatcc agactaaccc taattcaata gcatatgtta cccaacggga agcatatgct
     2461 atcgaattag ggttagtaaa agggtcctaa ggaacagcga tatctcccac cccatgagct
     2521 gtcacggttt tatttacatg gggtcaggat tccacgaggg tagtgaacca ttttagtcac
     2581 aagggcagtg gctgaagatc aaggagcggg cagtgaactc tcctgaatct tcgcctgctt
     2641 cttcattctc cttcgtttag ctaatagaat aactgctgag ttgtgaacag taaggtgtat
     2701 gtgaggtgct cgaaaacaag gtttcaggtg acgcccccag aataaaattt ggacgggggg
     2761 ttcagtggtg gcattgtgct atgacaccaa tataaccctc acaaacccct tgggcaataa
     2821 atactagtgt aggaatgaaa cattctgaat atctttaaca atagaaatcc atggggtggg
     2881 gacaagccgt aaagactgga tgtccatctc acacgaattt atggctatgg gcaacacata
     2941 atcctagtgc aatatgatac tggggttatt aagatgtgtc ccaggcaggg accaagacag
     3001 gtgaaccatg ttgttacact ctatttgtaa caaggggaaa gagagtggac gccgacagca
     3061 gcggactcca ctggttgtct ctaacacccc cgaaaattaa acggggctcc acgccaatgg
     3121 ggcccataaa caaagacaag tggccactct tttttttgaa attgtggagt gggggcacgc
     3181 gtcagccccc acacgccgcc ctgcggtttt ggactgtaaa ataagggtgt aataacttgg
     3241 ctgattgtaa ccccgctaac cactgcggtc aaaccacttg cccacaaaac cactaatggc
     3301 accccgggga atacctgcat aagtaggtgg gcgggccaag ataggggcgc gattgctgcg
     3361 atctggagga caaattacac acacttgcgc ctgagcgcca agcacagggt tgttggtcct
     3421 catattcacg aggtcgctga gagcacggtg ggctaatgtt gccatgggta gcatatacta
     3481 cccaaatatc tggatagcat atgctatcct aatctatatc tgggtagcat aggctatcct
     3541 aatctatatc tgggtagcat atgctatcct aatctatatc tgggtagtat atgctatcct
     3601 aatttatatc tgggtagcat aggctatcct aatctatatc tgggtagcat atgctatcct
     3661 aatctatatc tgggtagtat atgctatcct aatctgtatc cgggtagcat atgctatcct
     3721 aatagagatt agggtagtat atgctatcct aatttatatc tgggtagcat atactaccca
     3781 aatatctgga tagcatatgc tatcctaatc tatatctggg tagcatatgc tatcctaatc
     3841 tatatctggg tagcataggc tatcctaatc tatatctggg tagcatatgc tatcctaatc
     3901 tatatctggg tagtatatgc tatcctaatt tatatctggg tagcataggc tatcctaatc
     3961 tatatctggg tagcatatgc tatcctaatc tatatctggg tagtatatgc tatcctaatc
     4021 tgtatccggg tagcatatgc tatcctcatg ataagctgtc aaacatgaga attttcttga
     4081 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     4141 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4201 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4261 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     4321 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     4381 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     4441 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     4501 ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata
     4561 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     4621 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     4681 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     4741 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     4801 gacgagcgtg acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact
     4861 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     4921 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     4981 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     5041 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     5101 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     5161 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     5221 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     5281 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     5341 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     5401 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     5461 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     5521 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     5581 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     5641 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     5701 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     5761 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     5821 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     5881 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     5941 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     6001 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     6061 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     6121 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     6181 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     6241 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     6301 gaccatgatt acgccaagct ctagctagag gtcgaccaat tctcatgttt gacagcttat
     6361 catcgcagat ccgggcaacg ttgttgccat tgctgcaggc gcagaactgg taggtatgga
     6421 agatctatac attgaatcaa tattggcaat tagccatatt agtcattggt tatatagcat
     6481 aaatcaatat tggctattgg ccattgcata cgttgtatct atatcataat atgtacattt
     6541 atattggctc atgtccaata tgaccgccat gtt
//
