LOCUS       sequence-274732-        7935 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 7935)
  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 7935)
  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: D90_Ld_350740
FEATURES             Location/Qualifiers
     source          1..7935
                     /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             2713..2730
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2782..2837
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2836..2855)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5073..5091)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5159..5263
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5264..5332,5333..6124)
                     /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(5482..5501)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6295..6883
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6784..6803
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7037..7054
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7171..7192
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7207..7212,7213..7230,7231..7237)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7245..7261
                     /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     7250..7272
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7269..7285
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7269..7285
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7556..7935
                     /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 gccagcacgc cagcccccgc tcctggagaa
      841 tccccgtcta agaccgctct caatgcagaa gtctctgtct ctaccgctcc cccgccaact
      901 acatctagta gtgcggtgcc aagcggcagc gctaccgacc aacacagagg ggccgatgca
      961 ccttatagaa gcctgtatcc acaagttaag ctccgggcca caacccagga aaggccatat
     1021 aaacgaagag gctttgttgc cttcgaagaa gcagagacag gggtagagca acggcctaga
     1081 ctcgaagatg gctccgccca agttattgct cacctgccca ccgcagcaga aactggcggg
     1141 gccgcaattc tgcctggtca tagaagaagt cacgctcaaa cttcacttgc tactcaagag
     1201 ggcggctgcg ggcagttgag cgatgcagct gaaggcaatg aggcccaaca atttctggac
     1261 gaagtacaaa ggcaggtcga cgtagctaga catgctagga accaggccat ccctttcccc
     1321 cagaaacctg aagcgaattc cccagctttt cgaaggataa agagacacgc caaaatgaag
     1381 attgaggtgc ccgaccccga ctatcccacc ttcgctcgca aggacaaagt catgcaactg
     1441 cccccacccc acgaacatcc atgggttcgg aagaacacac ctataggacc ctttattgta
     1501 cacggggatg gtcagattca tcagacaggc ggcactggcc aggtggattt cgatgattcc
     1561 tctgtgaatg aaaagctgcc tcagtcctac cgcgggcttc agcaccggtc cacccttcaa
     1621 cggcaattgc ctcaggacaa tggaagggtg cttcaagagg ccgttataaa acacagcttc
     1681 acacttacgg gcagaggcct gtttgctact cgacggatag caaaagggga aaccatcatg
     1741 attgttcaga gtacggcacg caacgtagga gtaaagggag agcttcaacg tttggaagag
     1801 atgtgcacag acatactgat cgcttgtaga gatggggacg cccgtgcttg cgactacttg
     1861 catgattgga tcctgactgg acagccaagt tccctgctcg agcactggcc agccgcatcc
     1921 acttcccgag tcatggacgc catcggcggt gcggatgtgc ttagagcgct tgagctgcat
     1981 ccgatccaca ttgcgcgcat ggcagccatt attgatctga atagtttcct ggttgagtcc
     2041 agcttttccg aaaggaaagg catggcatac tttcccgaag cagggttcct gaaccactcc
     2101 tgcgctccga atgccactta tgacattatg cctgaacatg ttttccggga gacagattat
     2161 tacctggacg aagcaaccag ggcgacggca gcggaatccg gagaagaaga ggacgtgcag
     2221 ggtactgtga tggccgcaga tgatgcaggt acatcttcaa cagctgcgcc atgtccggca
     2281 cgtgccgctc agcccccagc cgcagtggga tatattagct cctcacaggt tctggtctgt
     2341 gacggcgtac ccgaaggaca aagacacgtt gcgttcttcg gtcatgaaga ccgactggcc
     2401 gcctaccccg ggctgacgga agccgatgct ccggtctact tgttttgttg tagagccgag
     2461 aaggaaatag aggctggtga ggaaattgtg attagctacg tgcccccgca gtggtcattt
     2521 gataatcgac agtacgtgct gcatgatcgc tacaggtttt ggtgtaagtg tccgaaatgc
     2581 agccccgtgc ttgataagaa gtacgccagg gtgcctggcg cgcccggcag cctgcaccac
     2641 atcctggacg cccagaagat gctgtggaac cacagggaca ggaacctgcc tcctctggcc
     2701 cctctgggcc cccaccacca tcaccaccat tgagtctaga cacgtgatta attaaggatc
     2761 ccccgacctc gacctctggc taataaagga aatttatttt cattgcaata gtgtgttgga
     2821 attttttgtg tctctcactc ggaaggacat atgggagggc aaatcatttg gtcgagatcc
     2881 ctcggagatc tctagctaga ggatcgatcc ccgccccgga cgaactaaac ctgactacga
     2941 catctctgcc ccttcttcgc ggggcagtgc atgtaatccc ttcagttggt tggtacaact
     3001 tgccaactgg gccctgttcc acatgtgaca cgggggggga ccaaacacaa aggggttctc
     3061 tgactgtagt tgacatcctt ataaatggat gtgcacattt gccaacactg agtggctttc
     3121 atcctggagc agactttgca gtctgtggac tgcaacacaa cattgccttt atgtgtaact
     3181 cttggctgaa gctcttacac caatgctggg ggacatgtac ctcccagggg cccaggaaga
     3241 ctacgggagg ctacaccaac gtcaatcaga ggggcctgtg tagctaccga taagcggacc
     3301 ctcaagaggg cattagcaat agtgtttata aggccccctt gttaacccta aacgggtagc
     3361 atatgcttcc cgggtagtag tatatactat ccagactaac cctaattcaa tagcatatgt
     3421 tacccaacgg gaagcatatg ctatcgaatt agggttagta aaagggtcct aaggaacagc
     3481 gatatctccc accccatgag ctgtcacggt tttatttaca tggggtcagg attccacgag
     3541 ggtagtgaac cattttagtc acaagggcag tggctgaaga tcaaggagcg ggcagtgaac
     3601 tctcctgaat cttcgcctgc ttcttcattc tccttcgttt agctaataga ataactgctg
     3661 agttgtgaac agtaaggtgt atgtgaggtg ctcgaaaaca aggtttcagg tgacgccccc
     3721 agaataaaat ttggacgggg ggttcagtgg tggcattgtg ctatgacacc aatataaccc
     3781 tcacaaaccc cttgggcaat aaatactagt gtaggaatga aacattctga atatctttaa
     3841 caatagaaat ccatggggtg gggacaagcc gtaaagactg gatgtccatc tcacacgaat
     3901 ttatggctat gggcaacaca taatcctagt gcaatatgat actggggtta ttaagatgtg
     3961 tcccaggcag ggaccaagac aggtgaacca tgttgttaca ctctatttgt aacaagggga
     4021 aagagagtgg acgccgacag cagcggactc cactggttgt ctctaacacc cccgaaaatt
     4081 aaacggggct ccacgccaat ggggcccata aacaaagaca agtggccact cttttttttg
     4141 aaattgtgga gtgggggcac gcgtcagccc ccacacgccg ccctgcggtt ttggactgta
     4201 aaataagggt gtaataactt ggctgattgt aaccccgcta accactgcgg tcaaaccact
     4261 tgcccacaaa accactaatg gcaccccggg gaatacctgc ataagtaggt gggcgggcca
     4321 agataggggc gcgattgctg cgatctggag gacaaattac acacacttgc gcctgagcgc
     4381 caagcacagg gttgttggtc ctcatattca cgaggtcgct gagagcacgg tgggctaatg
     4441 ttgccatggg tagcatatac tacccaaata tctggatagc atatgctatc ctaatctata
     4501 tctgggtagc ataggctatc ctaatctata tctgggtagc atatgctatc ctaatctata
     4561 tctgggtagt atatgctatc ctaatttata tctgggtagc ataggctatc ctaatctata
     4621 tctgggtagc atatgctatc ctaatctata tctgggtagt atatgctatc ctaatctgta
     4681 tccgggtagc atatgctatc ctaatagaga ttagggtagt atatgctatc ctaatttata
     4741 tctgggtagc atatactacc caaatatctg gatagcatat gctatcctaa tctatatctg
     4801 ggtagcatat gctatcctaa tctatatctg ggtagcatag gctatcctaa tctatatctg
     4861 ggtagcatat gctatcctaa tctatatctg ggtagtatat gctatcctaa tttatatctg
     4921 ggtagcatag gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg
     4981 ggtagtatat gctatcctaa tctgtatccg ggtagcatat gctatcctca tgataagctg
     5041 tcaaacatga gaattttctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt
     5101 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     5161 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     5221 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     5281 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     5341 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     5401 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     5461 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
     5521 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     5581 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     5641 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     5701 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     5761 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac
     5821 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     5881 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     5941 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     6001 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     6061 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     6121 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     6181 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     6241 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     6301 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     6361 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     6421 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     6481 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     6541 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     6601 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     6661 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6721 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6781 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6841 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6901 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     6961 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     7021 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     7081 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     7141 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
     7201 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
     7261 atttcacaca ggaaacagct atgaccatga ttacgccaag ctctagctag aggtcgacca
     7321 attctcatgt ttgacagctt atcatcgcag atccgggcaa cgttgttgcc attgctgcag
     7381 gcgcagaact ggtaggtatg gaagatctat acattgaatc aatattggca attagccata
     7441 ttagtcattg gttatatagc ataaatcaat attggctatt ggccattgca tacgttgtat
     7501 ctatatcata atatgtacat ttatattggc tcatgtccaa tatgaccgcc atgttgacat
     7561 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
     7621 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     7681 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     7741 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     7801 tatcatatgc caagtccgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     7861 tatgcccagt acatgacctt acgggacttt cctacttggc agtacatcta cgtattagtc
     7921 atcgctatta ccatg
//
