LOCUS       sequence-307590-        7353 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 7353)
  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 7353)
  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: D38_Ld_341960
FEATURES             Location/Qualifiers
     source          1..7353
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     201..225
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     706..725
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      748..757
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2131..2148
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2200..2255
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2254..2273)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4491..4509)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4577..4681
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4682..4750,4751..5542)
                     /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(4900..4919)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5713..6301
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6202..6221
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6455..6472
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6589..6610
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6625..6630,6631..6648,6649..6655)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6663..6679
                     /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     6668..6690
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6687..6703
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6687..6703
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        6974..7353
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta caccaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcctcac tctcttccgc
      241 atcgctgtct gcgagggcca gctgttgggc tcgcggttga ggacaaactc ttcgcggtct
      301 ttccagtact cttggatcgg aaacccgtcg gcctccgaac ggtactccgc caccgaggga
      361 cctgagcgag tccgcatcga ccggatcgga aaacctctcg agaaaggcgt ctaaccagtc
      421 acagtcgcaa ggtaggctga gcaccgtggc gggcggcagc gggtggcggt cggggttgtt
      481 tctggcggag gtgctgctga tgatgtaatt aaagtaggcg gtcttgagac ggcggatggt
      541 cgaggtgagg tgtggcaggc ttgagatcca gctgttgggg tgagtactcc ctctcaaaag
      601 cgggcattac ttctgcgcta agattgtcag tttccaaaaa cgaggaggat ttgatattca
      661 cctggcccga tctggccata cacttgagtg acaatgacat ccactttgcc tttctctcca
      721 caggtgtcca ctcccaggtc caagtttgcc accatggagt ttcagaccca ggtactcatg
      781 tccctgctgc tctgcatgtc tggtgcggcc gccacctacc cttacggcag atctagcgcc
      841 gtgaccgagc tgacccctgc cagcctgcac gccttcgtga acacccacaa gcccgtggtc
      901 atcctgttct acgccccttg gtgcggccac tgcaagcagt tccaccccga gtacgagaga
      961 ttcgccgaga gcgtgaaggg caccatcaga gtgggcgcca tcgacgccga caagaatgcc
     1021 gacatcggcc agcagttcgg cgtgcggggc ttccccacca tcaagtactg gaagtccggc
     1081 accaagagcg tgtccagcag ccaggaatac cagggccaga gaaccgccgc tgccctgcag
     1141 tctctggtgg tggaaggcat cagcagcagc aaagtgatga ccgtgaccac cgccgagcag
     1201 atgaagcagg ccgccaggga cgcccccaag aaaatgatcg gagtgctgct gagcgccaag
     1261 aacaaggtgc cccccatgtt ctccgtgatg ggcatgagcc ccaagctgaa agaactgccc
     1321 ctggtgttcg ccggcagcag ctcccctgag aagggcgtgg ccaaggccct gggagtgtcc
     1381 cagctgccca tgattggcgt gctgcggtac accccagccg agggggacgc cgaagagaag
     1441 ttcgagatgg tgccctatgc caagaaggcc attgcctacg agcctgtggc ccggttcttc
     1501 ctggactgcg ccgagaaaga gtgcagcgcc ggcaccgagg cccccatgaa ttccgtgtcc
     1561 ctggacgagg acgagagcca gcaccagacc cgggaagccg ccagacagtc tagcgtggcc
     1621 ctgcccgtgg aacccgtcga gtacacacct aaagtgatca gcctgttctg tgcccctcag
     1681 accagaaaga tcagaggcca gagccccctg tgcgtgttca gcctgagcga gcacgtgaac
     1741 ctggacaacg tgcaccagta cttcaagaac gaccccctgc tgttcttcga ggcctcccag
     1801 cacaaagagg aactgctcca gcagctgcag gccgactttg gcgtgcagat cgccagcacc
     1861 ggcgaggcca tgacaagcct ggccggctcc atcgtggtgc tgagacacag caagcccacc
     1921 accgtcagat acgccctgct ggaaaacatc gacagcgacg acgacctgca ggctgctctg
     1981 cagcggatcc tgaacggcga ggtgcacctg agatccgaga gagtgggagc cgctggcgcg
     2041 cccggcagcc tgcaccacat cctggacgcc cagaagatgc tgtggaacca cagggacagg
     2101 aacctgcctc ctctggcccc tctgggcccc caccaccatc accaccattg agtctagaca
     2161 cgtgattaat taaggatccc ccgacctcga cctctggcta ataaaggaaa tttattttca
     2221 ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg aaggacatat gggagggcaa
     2281 atcatttggt cgagatccct cggagatctc tagctagagg atcgatcccc gccccggacg
     2341 aactaaacct gactacgaca tctctgcccc ttcttcgcgg ggcagtgcat gtaatccctt
     2401 cagttggttg gtacaacttg ccaactgggc cctgttccac atgtgacacg gggggggacc
     2461 aaacacaaag gggttctctg actgtagttg acatccttat aaatggatgt gcacatttgc
     2521 caacactgag tggctttcat cctggagcag actttgcagt ctgtggactg caacacaaca
     2581 ttgcctttat gtgtaactct tggctgaagc tcttacacca atgctggggg acatgtacct
     2641 cccaggggcc caggaagact acgggaggct acaccaacgt caatcagagg ggcctgtgta
     2701 gctaccgata agcggaccct caagagggca ttagcaatag tgtttataag gcccccttgt
     2761 taaccctaaa cgggtagcat atgcttcccg ggtagtagta tatactatcc agactaaccc
     2821 taattcaata gcatatgtta cccaacggga agcatatgct atcgaattag ggttagtaaa
     2881 agggtcctaa ggaacagcga tatctcccac cccatgagct gtcacggttt tatttacatg
     2941 gggtcaggat tccacgaggg tagtgaacca ttttagtcac aagggcagtg gctgaagatc
     3001 aaggagcggg cagtgaactc tcctgaatct tcgcctgctt cttcattctc cttcgtttag
     3061 ctaatagaat aactgctgag ttgtgaacag taaggtgtat gtgaggtgct cgaaaacaag
     3121 gtttcaggtg acgcccccag aataaaattt ggacgggggg ttcagtggtg gcattgtgct
     3181 atgacaccaa tataaccctc acaaacccct tgggcaataa atactagtgt aggaatgaaa
     3241 cattctgaat atctttaaca atagaaatcc atggggtggg gacaagccgt aaagactgga
     3301 tgtccatctc acacgaattt atggctatgg gcaacacata atcctagtgc aatatgatac
     3361 tggggttatt aagatgtgtc ccaggcaggg accaagacag gtgaaccatg ttgttacact
     3421 ctatttgtaa caaggggaaa gagagtggac gccgacagca gcggactcca ctggttgtct
     3481 ctaacacccc cgaaaattaa acggggctcc acgccaatgg ggcccataaa caaagacaag
     3541 tggccactct tttttttgaa attgtggagt gggggcacgc gtcagccccc acacgccgcc
     3601 ctgcggtttt ggactgtaaa ataagggtgt aataacttgg ctgattgtaa ccccgctaac
     3661 cactgcggtc aaaccacttg cccacaaaac cactaatggc accccgggga atacctgcat
     3721 aagtaggtgg gcgggccaag ataggggcgc gattgctgcg atctggagga caaattacac
     3781 acacttgcgc ctgagcgcca agcacagggt tgttggtcct catattcacg aggtcgctga
     3841 gagcacggtg ggctaatgtt gccatgggta gcatatacta cccaaatatc tggatagcat
     3901 atgctatcct aatctatatc tgggtagcat aggctatcct aatctatatc tgggtagcat
     3961 atgctatcct aatctatatc tgggtagtat atgctatcct aatttatatc tgggtagcat
     4021 aggctatcct aatctatatc tgggtagcat atgctatcct aatctatatc tgggtagtat
     4081 atgctatcct aatctgtatc cgggtagcat atgctatcct aatagagatt agggtagtat
     4141 atgctatcct aatttatatc tgggtagcat atactaccca aatatctgga tagcatatgc
     4201 tatcctaatc tatatctggg tagcatatgc tatcctaatc tatatctggg tagcataggc
     4261 tatcctaatc tatatctggg tagcatatgc tatcctaatc tatatctggg tagtatatgc
     4321 tatcctaatt tatatctggg tagcataggc tatcctaatc tatatctggg tagcatatgc
     4381 tatcctaatc tatatctggg tagtatatgc tatcctaatc tgtatccggg tagcatatgc
     4441 tatcctcatg ataagctgtc aaacatgaga attttcttga agacgaaagg gcctcgtgat
     4501 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     4561 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     4621 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     4681 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     4741 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     4801 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     4861 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     4921 ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     4981 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     5041 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     5101 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     5161 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     5221 gcctgcagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     5281 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     5341 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     5401 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     5461 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     5521 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     5581 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     5641 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     5701 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     5761 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     5821 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     5881 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     5941 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     6001 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     6061 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     6121 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     6181 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     6241 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     6301 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     6361 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     6421 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     6481 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     6541 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
     6601 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
     6661 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct
     6721 ctagctagag gtcgaccaat tctcatgttt gacagcttat catcgcagat ccgggcaacg
     6781 ttgttgccat tgctgcaggc gcagaactgg taggtatgga agatctatac attgaatcaa
     6841 tattggcaat tagccatatt agtcattggt tatatagcat aaatcaatat tggctattgg
     6901 ccattgcata cgttgtatct atatcataat atgtacattt atattggctc atgtccaata
     6961 tgaccgccat gttgacattg attattgact agttattaat agtaatcaat tacggggtca
     7021 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
     7081 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
     7141 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     7201 ttggcagtac atcaagtgta tcatatgcca agtccgcccc ctattgacgt caatgacggt
     7261 aaatggcccg cctggcatta tgcccagtac atgaccttac gggactttcc tacttggcag
     7321 tacatctacg tattagtcat cgctattacc atg
//
