LOCUS       sequence-274722-        8145 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 8145)
  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 8145)
  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: D50_Ld_100670
FEATURES             Location/Qualifiers
     source          1..8145
                     /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             3303..3320
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3372..3427
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3426..3445)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5663..5681)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5749..5853
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5854..5922,5923..6714)
                     /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(6072..6091)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6885..7473
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7374..7393
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7627..7644
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7761..7782
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7797..7802,7803..7820,7821..7827)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7835..7851
                     /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     7840..7862
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7859..7875
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7859..7875
                     /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 gccaccgctg
     1201 ccgtggccac aacaacagcc agagcctacg accacgccgg catcacagtg gctggcgccc
     1261 tgctcgtggg ccagaacctg cagggaaagg caggcgccag ccggatcctg aaccccttcg
     1321 ccatctgcgc cgacttcgac accgccgacg tggaagatac caccctgctg atcggcggag
     1381 ccagctactt cttcagcttc aaccggtaca gcacctacct gggcttttgg tacggccagg
     1441 gcagcatgaa cctgaacagc ggccccatcg acaaagtgcg gctgaccggc gtgttcggct
     1501 gcgtgaccct gcggcccaac agcagcaatg gcctggtcac cagcaccgtg tactacgtgc
     1561 agaacgacgg catgctgtac tgggtgtcca acagcgtggt gtacctgacc caggtcaaac
     1621 acggcatcag cttcgtggac gtgaccgtgc acgacaacaa tgtgtatctg ctgagcaccc
     1681 agaacggcat ctaccagtgc ggcatcggcg ctggcggagc tgtcgtgggc agcgcctgta
     1741 cccagatcac cctgacaggc agcaccgagt tccaccagct gatccccatc agcagcgact
     1801 tccggggctt cgccgtgtcc agctccggca tcttcatcac ccccaccagc gacctgtatt
     1861 ggttcaacct gagcggcgcc tttatcgcca agagcgtggg cgtgacattc gtggatacca
     1921 agttcacctc cagccgggat accgccaacc ggggcacatc tgtgctgatg gccgccagca
     1981 cctccgccgt gtacaccgtg accacctccg acgccaccat cagctacgcc ctggtgtccg
     2041 gcaaagaagt gaagtcctgc aaccccgccc tgaacaacgt ggacagcgat accagcccca
     2101 ccttctgcgg aatcgcccgg atctaccccc tgtccaccga catggtgtat atgaccacag
     2161 gcggcgctag cgtcgtgcgg gccatcatcg tgtccaatac caccatctcc gacaccatca
     2221 cccggacccc cttccctgtg tacttcctgg acaacagctc catcatgccc ctgatcctgg
     2281 atggcatgaa ctacgagctg gtgtctaact ccaacatccc ctttccctac gtggccatca
     2341 atcacagcac ccccaaggtg gacgactcca cctgggacac aatcttctcc gtggatgtgt
     2401 caaaccggtt ctttagcaca gtgtccagag ccgccgtgat cggcacccct ttcatgagca
     2461 gcctgcacgg actgcaggcc tactacaacc ggaccaacca catcctgttc ggcgacccca
     2521 acgtgctgcc catgtgcaac ctgaccaaga tgcagatgat cgagcgggcc gtggctgccg
     2581 atgccagagc tgccctgcag tacccctaca tctacaccag caaggcccag aacttcaccg
     2641 tgaacgccca ccccaatctg acactgctga agctgctgat gccctaccct ttcggcgaga
     2701 ttctgaacga gagcggcttc ttcgagaaca ccaccacccc cgcctccctg gccaatgtgc
     2761 acttcaacac cacaatgctg gccgctgtgc ggaacgccta cacccccgat ttcgtgtacg
     2821 actgcatctt cgccggcaac gccttcccct tccacattct gaccgccgct cagcagcagc
     2881 tcgtgcggtg gatcatctat acagccatcc aggaacagct ggccaaatgc gccgagaaca
     2941 gccccagcta caccggcagc ggcagctcct ccagcgacag ccacgacgat atggtgcccg
     3001 gctgtgtgcc cagagtgggc atcgataacc tgacagagct ggtcatgccc ggcatgcctt
     3061 acagcaacta caacatcacc gtgttcatcc ccgagtccct gcactacaac ttcagcatct
     3121 cccggtgcct ggacggcacc gactggacca acgtgacaga ttacctgcag aatgccacca
     3181 caaccagaac ccggaagtgt ggcacaggcg cgcccggcag cctgcaccac atcctggacg
     3241 cccagaagat gctgtggaac cacagggaca ggaacctgcc tcctctggcc cctctgggcc
     3301 cccaccacca tcaccaccat tgagtctaga cacgtgatta attaaggatc ccccgacctc
     3361 gacctctggc taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg
     3421 tctctcactc ggaaggacat atgggagggc aaatcatttg gtcgagatcc ctcggagatc
     3481 tctagctaga ggatcgatcc ccgccccgga cgaactaaac ctgactacga catctctgcc
     3541 ccttcttcgc ggggcagtgc atgtaatccc ttcagttggt tggtacaact tgccaactgg
     3601 gccctgttcc acatgtgaca cgggggggga ccaaacacaa aggggttctc tgactgtagt
     3661 tgacatcctt ataaatggat gtgcacattt gccaacactg agtggctttc atcctggagc
     3721 agactttgca gtctgtggac tgcaacacaa cattgccttt atgtgtaact cttggctgaa
     3781 gctcttacac caatgctggg ggacatgtac ctcccagggg cccaggaaga ctacgggagg
     3841 ctacaccaac gtcaatcaga ggggcctgtg tagctaccga taagcggacc ctcaagaggg
     3901 cattagcaat agtgtttata aggccccctt gttaacccta aacgggtagc atatgcttcc
     3961 cgggtagtag tatatactat ccagactaac cctaattcaa tagcatatgt tacccaacgg
     4021 gaagcatatg ctatcgaatt agggttagta aaagggtcct aaggaacagc gatatctccc
     4081 accccatgag ctgtcacggt tttatttaca tggggtcagg attccacgag ggtagtgaac
     4141 cattttagtc acaagggcag tggctgaaga tcaaggagcg ggcagtgaac tctcctgaat
     4201 cttcgcctgc ttcttcattc tccttcgttt agctaataga ataactgctg agttgtgaac
     4261 agtaaggtgt atgtgaggtg ctcgaaaaca aggtttcagg tgacgccccc agaataaaat
     4321 ttggacgggg ggttcagtgg tggcattgtg ctatgacacc aatataaccc tcacaaaccc
     4381 cttgggcaat aaatactagt gtaggaatga aacattctga atatctttaa caatagaaat
     4441 ccatggggtg gggacaagcc gtaaagactg gatgtccatc tcacacgaat ttatggctat
     4501 gggcaacaca taatcctagt gcaatatgat actggggtta ttaagatgtg tcccaggcag
     4561 ggaccaagac aggtgaacca tgttgttaca ctctatttgt aacaagggga aagagagtgg
     4621 acgccgacag cagcggactc cactggttgt ctctaacacc cccgaaaatt aaacggggct
     4681 ccacgccaat ggggcccata aacaaagaca agtggccact cttttttttg aaattgtgga
     4741 gtgggggcac gcgtcagccc ccacacgccg ccctgcggtt ttggactgta aaataagggt
     4801 gtaataactt ggctgattgt aaccccgcta accactgcgg tcaaaccact tgcccacaaa
     4861 accactaatg gcaccccggg gaatacctgc ataagtaggt gggcgggcca agataggggc
     4921 gcgattgctg cgatctggag gacaaattac acacacttgc gcctgagcgc caagcacagg
     4981 gttgttggtc ctcatattca cgaggtcgct gagagcacgg tgggctaatg ttgccatggg
     5041 tagcatatac tacccaaata tctggatagc atatgctatc ctaatctata tctgggtagc
     5101 ataggctatc ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt
     5161 atatgctatc ctaatttata tctgggtagc ataggctatc ctaatctata tctgggtagc
     5221 atatgctatc ctaatctata tctgggtagt atatgctatc ctaatctgta tccgggtagc
     5281 atatgctatc ctaatagaga ttagggtagt atatgctatc ctaatttata tctgggtagc
     5341 atatactacc caaatatctg gatagcatat gctatcctaa tctatatctg ggtagcatat
     5401 gctatcctaa tctatatctg ggtagcatag gctatcctaa tctatatctg ggtagcatat
     5461 gctatcctaa tctatatctg ggtagtatat gctatcctaa tttatatctg ggtagcatag
     5521 gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagtatat
     5581 gctatcctaa tctgtatccg ggtagcatat gctatcctca tgataagctg tcaaacatga
     5641 gaattttctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     5701 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     5761 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     5821 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     5881 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     5941 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     6001 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     6061 tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc
     6121 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     6181 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     6241 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     6301 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     6361 gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac aacgttgcgc
     6421 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     6481 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     6541 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     6601 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     6661 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     6721 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     6781 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     6841 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     6901 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     6961 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     7021 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     7081 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     7141 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     7201 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     7261 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     7321 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     7381 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     7441 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     7501 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     7561 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     7621 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     7681 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     7741 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     7801 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     7861 ggaaacagct atgaccatga ttacgccaag ctctagctag aggtcgacca attctcatgt
     7921 ttgacagctt atcatcgcag atccgggcaa cgttgttgcc attgctgcag gcgcagaact
     7981 ggtaggtatg gaagatctat acattgaatc aatattggca attagccata ttagtcattg
     8041 gttatatagc ataaatcaat attggctatt ggccattgca tacgttgtat ctatatcata
     8101 atatgtacat ttatattggc tcatgtccaa tatgaccgcc atgtt
//
