LOCUS       sequence-262301-        8082 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 8082)
  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 8082)
  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: D6_Ld_100150
FEATURES             Location/Qualifiers
     source          1..8082
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        7..386
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        387..590
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     541..561
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     587..611
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     1092..1111
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1134..1143
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3246..3263
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3315..3370
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3369..3388)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5606..5624)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5692..5796
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5797..5865,5866..6657)
                     /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(6015..6034)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6828..7416
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7317..7336
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7570..7587
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7704..7725
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7740..7745,7746..7763,7764..7770)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7778..7794
                     /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     7783..7805
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7802..7818
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7802..7818
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 catgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
       61 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      121 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      181 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      241 tacatcaagt gtatcatatg ccaagtccgc cccctattga cgtcaatgac ggtaaatggc
      301 ccgcctggca ttatgcccag tacatgacct tacgggactt tcctacttgg cagtacatct
      361 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacacc aatgggcgtg
      421 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      481 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taataacccc gccccgttga
      541 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga
      601 accgtcagat cctcactctc ttccgcatcg ctgtctgcga gggccagctg ttgggctcgc
      661 ggttgaggac aaactcttcg cggtctttcc agtactcttg gatcggaaac ccgtcggcct
      721 ccgaacggta ctccgccacc gagggacctg agcgagtccg catcgaccgg atcggaaaac
      781 ctctcgagaa aggcgtctaa ccagtcacag tcgcaaggta ggctgagcac cgtggcgggc
      841 ggcagcgggt ggcggtcggg gttgtttctg gcggaggtgc tgctgatgat gtaattaaag
      901 taggcggtct tgagacggcg gatggtcgag gtgaggtgtg gcaggcttga gatccagctg
      961 ttggggtgag tactccctct caaaagcggg cattacttct gcgctaagat tgtcagtttc
     1021 caaaaacgag gaggatttga tattcacctg gcccgatctg gccatacact tgagtgacaa
     1081 tgacatccac tttgcctttc tctccacagg tgtccactcc caggtccaag tttgccacca
     1141 tggagtttca gacccaggta ctcatgtccc tgctgctctg catgtctggt gcggccgcca
     1201 ccaaccctct gcagagcaga cacatgcagc tgacccagaa cgccgtgaag aagtccgagg
     1261 ccaacctggt caactgccgg gaagtgaacg ccgacctgaa gtccggcgga ggcgtgaacg
     1321 ccgcccaggc cattgccgag atgcggcggc agcgcgagga actgatgaac atcgtggccc
     1381 tggaacgcga gcgggtggtg tccgctagaa gcctgctgca ggtctgcgag gacgagctgg
     1441 ccagcgacct gagcgtgctg ttcggcgtgg ccgaccacaa cttcaccgcc cggatcagaa
     1501 gcctggaaga gaagcggaag cacctggaag gcctgcacag catgctgaac accaccccct
     1561 tcggcgccgt ggaactgcgg cggtctagcg agattagagc cctgcaggcc gccctgctgc
     1621 acgagatgcg ggccagcaag aagaaagccg agaacggcgt ggccaacggc gaggcctgca
     1681 ccaagaccag cgacaagtac agcgtggtgt tcgacatcgg cagcaccggc aaccgggtgc
     1741 acgtgtacaa gtaccgggtg gcccctgcca cccatacagc cgccgctgcc ggctccgagc
     1801 tgagcgacat cgacctggtg gaagaactgt tcgagctgaa ccacaaggcc ctgagcgagc
     1861 tggacaaccc cgtgcaggat gcccctgagg ccctgtggga gctgttcatg aaggccaagt
     1921 acttcgtgcc cgccgagctg cacgcctgca ccgccgtgga attcaaggcc acagccggcc
     1981 tgcggatgct gggcatggaa aaggccaccg agatcctgga cggcatccgg gccagatacc
     2041 ggaacgagac attctggctg cggggcaacg cccccgtgcg gatcctggat gcctgcgaag
     2101 agggccccat ggcctggctg accgtgaact acctgctggg cgtgttcagc cggggcacca
     2161 aggccactgc cagcaccgtg gccgtgatcg atctgggcgg aggcagcacc cagatcgtgt
     2221 tcgagcctgg cgagagcgcc ttccacggca tgcggaccga cctgagatac agcgccaccc
     2281 tgggcagcag aagcgtgtcc gcctaccagc acagctacga gggctatggc ctgcacgccg
     2341 ccacaaaaga gctgctgttc catattcagg gcaagagcca ggaaaagcct ggcggcggaa
     2401 ccaccaccag caccgccacc accaccacaa ccactacccc cgccaacagc ggcgataagg
     2461 ccctgcccgt gtggaacgtg ctgggcaatc tgggcgccga cggcagcagc gagagggacg
     2521 gaatcgtgac caaacgggcc cctcccatgc cccctccacc tctgcctgat gccgaggccg
     2581 tggaagcctt cccttgcttc gccgtgggct acgaggaccc cctgggcgtg aagaacatca
     2641 agaagaacaa cgccggcgag cccgccatgc cccccaattt ccaggcctgc gccaacctgt
     2701 tccgggaccg gctgctgaag cctgtgggcc tgacatgcga ggccgccaac tgtggaatcg
     2761 ccggcgtgat gcagcccccc ctgaccaatt tcaccggcga gatctacgtg ttcagcttca
     2821 tcttcgacct gctggccctg gccaacagca gcctggtgcc tgctggcgcc gctgtgtccc
     2881 gcgagaagtt cgaagtgaag ctgcccgacc tggccacaat cgccgagggc cactgtgccg
     2941 ccttcagcct gaccagaatt gccgaggcca ccgccaaaga gggcctgggc agcctgaagc
     3001 ccgagtacga gtgcatgtac tacagctacg tgtacgccct gctgagatac ggctacgagg
     3061 tgcccgagga ccgggtgctg cacgtggtca agaagatccg gggctacgag acagcctggt
     3121 ccctgggcgc cagcctgctg agcctgacag gcgcgcccgg cagcctgcac cacatcctgg
     3181 acgcccagaa gatgctgtgg aaccacaggg acaggaacct gcctcctctg gcccctctgg
     3241 gcccccacca ccatcaccac cattgagtct agacacgtga ttaattaagg atcccccgac
     3301 ctcgacctct ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt
     3361 gtgtctctca ctcggaagga catatgggag ggcaaatcat ttggtcgaga tccctcggag
     3421 atctctagct agaggatcga tccccgcccc ggacgaacta aacctgacta cgacatctct
     3481 gccccttctt cgcggggcag tgcatgtaat cccttcagtt ggttggtaca acttgccaac
     3541 tgggccctgt tccacatgtg acacgggggg ggaccaaaca caaaggggtt ctctgactgt
     3601 agttgacatc cttataaatg gatgtgcaca tttgccaaca ctgagtggct ttcatcctgg
     3661 agcagacttt gcagtctgtg gactgcaaca caacattgcc tttatgtgta actcttggct
     3721 gaagctctta caccaatgct gggggacatg tacctcccag gggcccagga agactacggg
     3781 aggctacacc aacgtcaatc agaggggcct gtgtagctac cgataagcgg accctcaaga
     3841 gggcattagc aatagtgttt ataaggcccc cttgttaacc ctaaacgggt agcatatgct
     3901 tcccgggtag tagtatatac tatccagact aaccctaatt caatagcata tgttacccaa
     3961 cgggaagcat atgctatcga attagggtta gtaaaagggt cctaaggaac agcgatatct
     4021 cccaccccat gagctgtcac ggttttattt acatggggtc aggattccac gagggtagtg
     4081 aaccatttta gtcacaaggg cagtggctga agatcaagga gcgggcagtg aactctcctg
     4141 aatcttcgcc tgcttcttca ttctccttcg tttagctaat agaataactg ctgagttgtg
     4201 aacagtaagg tgtatgtgag gtgctcgaaa acaaggtttc aggtgacgcc cccagaataa
     4261 aatttggacg gggggttcag tggtggcatt gtgctatgac accaatataa ccctcacaaa
     4321 ccccttgggc aataaatact agtgtaggaa tgaaacattc tgaatatctt taacaataga
     4381 aatccatggg gtggggacaa gccgtaaaga ctggatgtcc atctcacacg aatttatggc
     4441 tatgggcaac acataatcct agtgcaatat gatactgggg ttattaagat gtgtcccagg
     4501 cagggaccaa gacaggtgaa ccatgttgtt acactctatt tgtaacaagg ggaaagagag
     4561 tggacgccga cagcagcgga ctccactggt tgtctctaac acccccgaaa attaaacggg
     4621 gctccacgcc aatggggccc ataaacaaag acaagtggcc actctttttt ttgaaattgt
     4681 ggagtggggg cacgcgtcag cccccacacg ccgccctgcg gttttggact gtaaaataag
     4741 ggtgtaataa cttggctgat tgtaaccccg ctaaccactg cggtcaaacc acttgcccac
     4801 aaaaccacta atggcacccc ggggaatacc tgcataagta ggtgggcggg ccaagatagg
     4861 ggcgcgattg ctgcgatctg gaggacaaat tacacacact tgcgcctgag cgccaagcac
     4921 agggttgttg gtcctcatat tcacgaggtc gctgagagca cggtgggcta atgttgccat
     4981 gggtagcata tactacccaa atatctggat agcatatgct atcctaatct atatctgggt
     5041 agcataggct atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt
     5101 agtatatgct atcctaattt atatctgggt agcataggct atcctaatct atatctgggt
     5161 agcatatgct atcctaatct atatctgggt agtatatgct atcctaatct gtatccgggt
     5221 agcatatgct atcctaatag agattagggt agtatatgct atcctaattt atatctgggt
     5281 agcatatact acccaaatat ctggatagca tatgctatcc taatctatat ctgggtagca
     5341 tatgctatcc taatctatat ctgggtagca taggctatcc taatctatat ctgggtagca
     5401 tatgctatcc taatctatat ctgggtagta tatgctatcc taatttatat ctgggtagca
     5461 taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagta
     5521 tatgctatcc taatctgtat ccgggtagca tatgctatcc tcatgataag ctgtcaaaca
     5581 tgagaatttt cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     5641 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     5701 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     5761 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     5821 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     5881 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     5941 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     6001 acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa
     6061 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     6121 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     6181 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     6241 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     6301 gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg
     6361 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     6421 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     6481 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     6541 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     6601 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     6661 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     6721 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     6781 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     6841 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     6901 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     6961 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     7021 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     7081 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     7141 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     7201 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     7261 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     7321 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     7381 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     7441 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     7501 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     7561 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     7621 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     7681 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     7741 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     7801 acaggaaaca gctatgacca tgattacgcc aagctctagc tagaggtcga ccaattctca
     7861 tgtttgacag cttatcatcg cagatccggg caacgttgtt gccattgctg caggcgcaga
     7921 actggtaggt atggaagatc tatacattga atcaatattg gcaattagcc atattagtca
     7981 ttggttatat agcataaatc aatattggct attggccatt gcatacgttg tatctatatc
     8041 ataatatgta catttatatt ggctcatgtc caatatgacc gc
//
