LOCUS       sequence-274884-        7383 bp    DNA     circular SYN 15-APR-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 7383)
  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 7383)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: D60_Ld_343410
FEATURES             Location/Qualifiers
     source          1..7383
                     /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             2161..2178
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2230..2285
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2284..2303)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4521..4539)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4607..4711
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4712..4780,4781..5572)
                     /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(4930..4949)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5743..6331
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6232..6251
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6485..6502
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6619..6640
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6655..6660,6661..6678,6679..6685)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6693..6709
                     /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     6698..6720
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6717..6733
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6717..6733
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7004..7383
                     /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 gcccatcagg tggccctgat cagcgacatc
      841 cactacgacc ccctgtacgg caccagcaga gccctgaagt gcaccagcga cagcagccct
      901 atgtacggca tgcagggctg cgacagctcc ctgcagctga tggccagaac cctggccgac
      961 gtgtccgccc agaaccccag cctggtgctg tacaccggcg actggcagcg gcacaagttc
     1021 gccaacagct ctctggcccc tgccgccatc ttcaaggaca tgagcgagcg gttccacaac
     1081 gtgaccgtgg acggctctct gggcgccgtg gccttttctg gcgccatggg caacaacgac
     1141 gtggtgcccg actactacta cagcctggaa gataccgtgt ccgaggaaga actggcccac
     1201 cgggtggccg ccatgagagg cgctgctctg ctgtctgatg ccgaggccag cgtgatgcgg
     1261 aactgcggct actacaccca catgatggac agcgtgcacg tgatcgtgct gcacaccctg
     1321 ctgtgggccc acgagctgca gccccccctg cccagcaatc tgagcgaccc ctgcaaccag
     1381 ttcagcttcc tgcggagcga gctggccaaa gtgcgggctg ccaacaagcg ggccatcatc
     1441 atgggccaca tcccccctgg cctgaacacc tacaacgtgc tgaagcgggg cttcggcagc
     1501 gccgctggcg acatgttctg gaaagagcag tacgaggcca cctacaacag catcatcaga
     1561 gagtacaagg acctgctggt ggtgcagctg ttcggccaca cacacatgtt caagctgctg
     1621 accatgcccc ggaacgacgt gctgggcatc gtggtgccca gcatcagccc catcttcggc
     1681 aaccacccca gctacctgat ggccaacttc agcgagacat gggagctgga agatgcccag
     1741 agccggtaca ccatcggcga gggcgtgttc cacagcggcc tgagcgccaa agaggtgctg
     1801 agcctgaacg ccggcctgca cagcgtgtca gacgtgcgcg ctgccatcac cctgctggcc
     1861 accaacgata ccatgtggga ccggtttctg accgtgttct gtggcggcga gaagcggagc
     1921 caggtgttcc cccaccggaa gtgcgacaag cagtgtcggt acatcatcgt gtgcagcatg
     1981 ctggaaaaca accacaccta catccagaga tgcgtggcca actactccct gctgcctggc
     2041 cccagccagg acaccagaac cacaggcgcg cccggcagcc tgcaccacat cctggacgcc
     2101 cagaagatgc tgtggaacca cagggacagg aacctgcctc ctctggcccc tctgggcccc
     2161 caccaccatc accaccattg agtctagaca cgtgattaat taaggatccc ccgacctcga
     2221 cctctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc
     2281 tctcactcgg aaggacatat gggagggcaa atcatttggt cgagatccct cggagatctc
     2341 tagctagagg atcgatcccc gccccggacg aactaaacct gactacgaca tctctgcccc
     2401 ttcttcgcgg ggcagtgcat gtaatccctt cagttggttg gtacaacttg ccaactgggc
     2461 cctgttccac atgtgacacg gggggggacc aaacacaaag gggttctctg actgtagttg
     2521 acatccttat aaatggatgt gcacatttgc caacactgag tggctttcat cctggagcag
     2581 actttgcagt ctgtggactg caacacaaca ttgcctttat gtgtaactct tggctgaagc
     2641 tcttacacca atgctggggg acatgtacct cccaggggcc caggaagact acgggaggct
     2701 acaccaacgt caatcagagg ggcctgtgta gctaccgata agcggaccct caagagggca
     2761 ttagcaatag tgtttataag gcccccttgt taaccctaaa cgggtagcat atgcttcccg
     2821 ggtagtagta tatactatcc agactaaccc taattcaata gcatatgtta cccaacggga
     2881 agcatatgct atcgaattag ggttagtaaa agggtcctaa ggaacagcga tatctcccac
     2941 cccatgagct gtcacggttt tatttacatg gggtcaggat tccacgaggg tagtgaacca
     3001 ttttagtcac aagggcagtg gctgaagatc aaggagcggg cagtgaactc tcctgaatct
     3061 tcgcctgctt cttcattctc cttcgtttag ctaatagaat aactgctgag ttgtgaacag
     3121 taaggtgtat gtgaggtgct cgaaaacaag gtttcaggtg acgcccccag aataaaattt
     3181 ggacgggggg ttcagtggtg gcattgtgct atgacaccaa tataaccctc acaaacccct
     3241 tgggcaataa atactagtgt aggaatgaaa cattctgaat atctttaaca atagaaatcc
     3301 atggggtggg gacaagccgt aaagactgga tgtccatctc acacgaattt atggctatgg
     3361 gcaacacata atcctagtgc aatatgatac tggggttatt aagatgtgtc ccaggcaggg
     3421 accaagacag gtgaaccatg ttgttacact ctatttgtaa caaggggaaa gagagtggac
     3481 gccgacagca gcggactcca ctggttgtct ctaacacccc cgaaaattaa acggggctcc
     3541 acgccaatgg ggcccataaa caaagacaag tggccactct tttttttgaa attgtggagt
     3601 gggggcacgc gtcagccccc acacgccgcc ctgcggtttt ggactgtaaa ataagggtgt
     3661 aataacttgg ctgattgtaa ccccgctaac cactgcggtc aaaccacttg cccacaaaac
     3721 cactaatggc accccgggga atacctgcat aagtaggtgg gcgggccaag ataggggcgc
     3781 gattgctgcg atctggagga caaattacac acacttgcgc ctgagcgcca agcacagggt
     3841 tgttggtcct catattcacg aggtcgctga gagcacggtg ggctaatgtt gccatgggta
     3901 gcatatacta cccaaatatc tggatagcat atgctatcct aatctatatc tgggtagcat
     3961 aggctatcct aatctatatc tgggtagcat atgctatcct aatctatatc tgggtagtat
     4021 atgctatcct aatttatatc tgggtagcat aggctatcct aatctatatc tgggtagcat
     4081 atgctatcct aatctatatc tgggtagtat atgctatcct aatctgtatc cgggtagcat
     4141 atgctatcct aatagagatt agggtagtat atgctatcct aatttatatc tgggtagcat
     4201 atactaccca aatatctgga tagcatatgc tatcctaatc tatatctggg tagcatatgc
     4261 tatcctaatc tatatctggg tagcataggc tatcctaatc tatatctggg tagcatatgc
     4321 tatcctaatc tatatctggg tagtatatgc tatcctaatt tatatctggg tagcataggc
     4381 tatcctaatc tatatctggg tagcatatgc tatcctaatc tatatctggg tagtatatgc
     4441 tatcctaatc tgtatccggg tagcatatgc tatcctcatg ataagctgtc aaacatgaga
     4501 attttcttga agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     4561 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     4621 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     4681 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     4741 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     4801 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     4861 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     4921 taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg
     4981 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     5041 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     5101 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     5161 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     5221 cataccaaac gacgagcgtg acaccacgat gcctgcagca atggcaacaa cgttgcgcaa
     5281 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     5341 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     5401 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     5461 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     5521 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     5581 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     5641 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     5701 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5761 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     5821 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5881 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     5941 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     6001 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     6061 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6121 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6181 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6241 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6301 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     6361 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     6421 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     6481 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     6541 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     6601 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
     6661 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
     6721 aaacagctat gaccatgatt acgccaagct ctagctagag gtcgaccaat tctcatgttt
     6781 gacagcttat catcgcagat ccgggcaacg ttgttgccat tgctgcaggc gcagaactgg
     6841 taggtatgga agatctatac attgaatcaa tattggcaat tagccatatt agtcattggt
     6901 tatatagcat aaatcaatat tggctattgg ccattgcata cgttgtatct atatcataat
     6961 atgtacattt atattggctc atgtccaata tgaccgccat gttgacattg attattgact
     7021 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     7081 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     7141 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     7201 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     7261 agtccgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     7321 atgaccttac gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     7381 atg
//
