LOCUS       sequence-262337-        8502 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 8502)
  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 8502)
  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: D44_Ld_180090
FEATURES             Location/Qualifiers
     source          1..8502
                     /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             3666..3683
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3735..3790
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3789..3808)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6026..6044)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6112..6216
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6217..6285,6286..7077)
                     /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(6435..6454)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7248..7836
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7737..7756
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7990..8007
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8124..8145
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8160..8165,8166..8183,8184..8190)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8198..8214
                     /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     8203..8225
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8222..8238
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8222..8238
                     /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 ccgtgaccgt ggtcacaaca gaaggcggcc tggccctgga agccatcggc taccccaacg
     1261 acgtcgtgcg gctgaccgcc aagcccggcg acaagtggac ctaccagccc gacggcatca
     1321 tcctgcccga cgccgtggct ctgcaggcca ccaaaggcgc ctccgacgcc cagtggtcca
     1381 tcccccacgg caagtgcatg ctgaccatca gcatggcccc tccccacgtg cgcttcgagt
     1441 tcagctgtca cggcagcagc ctggtggaag tgaccgcctt cgccgatgcc atggctgccc
     1501 ccgaagtgaa cttcaccttc cctgccgccc agactatgta cggactggcc gagcacgccg
     1561 ccgacctgcc tctgagaggc ggcaacgtgt acgagatgta caacaccgac accttccagt
     1621 acagcgtgaa cagcaccgag gccctgtacg gcgtgatccc cttcatcatg gcctacgccc
     1681 ccaagtctac ctgcggcgtg ctgttcctga accccagcga gacaaacgtg ggcgtgtccg
     1741 ccgatagcgc cgctcccagc tgccagtgga agcctgaagt gggcgccatc gacatctttt
     1801 ttctgcccgg acccaccccc accaaggtcc agcagcagca tgccgccctg accggcgcca
     1861 cagtgatgcc cccttacttc agcctgggcc tgcaccagtg ccggtggaac tacctgaaca
     1921 ccaaggactg cctgagcgtg gacgagggct tcgacaccca caacatgccc tacgacgtgc
     1981 tgtggctgga catcgagcac accgataaga agaagtactt tacctgggac ccctacacct
     2041 ttcccgaccc caaggccctg acagacgccc tggccagcaa gggcagaaag ctggtcaccg
     2101 tgcgggaccc tcacgtgaag cgggatgagg gctactacgt gcacaacgag gcccagaaag
     2161 gcggctacta tgtgaaggac gccagcggcg aggactacgt gggcaagtgt tggcccggca
     2221 gctccagctg gcccgacttc ttcaacaccc ggaccagagt gtggtacagc cagttcttcc
     2281 acgacgacag ataccctggc ggcagccggg acatccacac ctgggtggac atgaacgagc
     2341 ccagcgtgtt cggcggcgag cggggcacaa tgcctaagac cgccgtgcac agcctggaca
     2401 acggccacac cgtggaacac agattcgtgc ataacgccta cagcttctat agcgtgcagg
     2461 cagtgcacaa gggcatgctc gaggctggcg gccctaacac cgcccctgag cggcccttta
     2521 tcctgaccag aagcttcttc agcggctccc agagatacgc cgccatgtgg accggcgata
     2581 acatggccag atgggaccac ctggaaaaca gcatccccga gctgctgagc ctgagcatct
     2641 ccaactaccc tttctgcggc tgcgatatcg gcggattctt cttcgacccc gaagaggaac
     2701 tgttcgtgcg gtggatgcag gccggggtgt tcgtcccttt ctaccgggcc cacagccatc
     2761 tggacaccaa gcggagagag ccctggacct tcagcgccga agcccagtct ctcgtgcgca
     2821 gcgccctcgc cctgcgctac gccatgctgc cttacctgta cacaaccttc taccacgccc
     2881 acaccgaggg caacaccatc atgcggcccc tgttctacga gttcccccgg cagagcgagc
     2941 tgagagaggt gcagaacacc tacctgttcg gccccagcat cctcgtccgg cccgtcgtga
     3001 agcctagcgt gaccgaagtc acagtgcccc tgcccaaaga ggccctctgg tacaactact
     3061 tctccggcga gctggccgtg ggccagcaca ccatgcccgt ggacaaggac accatcccca
     3121 tgtttctgag gggcggacac atcgtgccca tgaagctgag actgcggcgg agcagcttcg
     3181 ccgccagact ggaccctttc accctgtttg tggccctgaa tgcccagggc aactcctacg
     3241 gcgatctgta catcgacgac ggcaccacct acgactacaa aaagggcgcc ttcgtgcacc
     3301 gggccttcag ctacagcaac caggtgctgc agaactccgc ctaccctaac agccccggca
     3361 ccagcggcct gtacaacgcc gtgaatgccg tggaaagagt ggtcatctac ggctatgtgg
     3421 gaaaggtgtc caaggccgtg ctgaataccc gggtggacga aaccgaggtg gccacccccc
     3481 tggaattcga gcagctgggc cagacagtgg tcgtccggaa gccttctgtg ctggtggccg
     3541 ccgattggat catcagcttc gagaacgaag gcgcgcccgg cagcctgcac cacatcctgg
     3601 acgcccagaa gatgctgtgg aaccacaggg acaggaacct gcctcctctg gcccctctgg
     3661 gcccccacca ccatcaccac cattgagtct agacacgtga ttaattaagg atcccccgac
     3721 ctcgacctct ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt
     3781 gtgtctctca ctcggaagga catatgggag ggcaaatcat ttggtcgaga tccctcggag
     3841 atctctagct agaggatcga tccccgcccc ggacgaacta aacctgacta cgacatctct
     3901 gccccttctt cgcggggcag tgcatgtaat cccttcagtt ggttggtaca acttgccaac
     3961 tgggccctgt tccacatgtg acacgggggg ggaccaaaca caaaggggtt ctctgactgt
     4021 agttgacatc cttataaatg gatgtgcaca tttgccaaca ctgagtggct ttcatcctgg
     4081 agcagacttt gcagtctgtg gactgcaaca caacattgcc tttatgtgta actcttggct
     4141 gaagctctta caccaatgct gggggacatg tacctcccag gggcccagga agactacggg
     4201 aggctacacc aacgtcaatc agaggggcct gtgtagctac cgataagcgg accctcaaga
     4261 gggcattagc aatagtgttt ataaggcccc cttgttaacc ctaaacgggt agcatatgct
     4321 tcccgggtag tagtatatac tatccagact aaccctaatt caatagcata tgttacccaa
     4381 cgggaagcat atgctatcga attagggtta gtaaaagggt cctaaggaac agcgatatct
     4441 cccaccccat gagctgtcac ggttttattt acatggggtc aggattccac gagggtagtg
     4501 aaccatttta gtcacaaggg cagtggctga agatcaagga gcgggcagtg aactctcctg
     4561 aatcttcgcc tgcttcttca ttctccttcg tttagctaat agaataactg ctgagttgtg
     4621 aacagtaagg tgtatgtgag gtgctcgaaa acaaggtttc aggtgacgcc cccagaataa
     4681 aatttggacg gggggttcag tggtggcatt gtgctatgac accaatataa ccctcacaaa
     4741 ccccttgggc aataaatact agtgtaggaa tgaaacattc tgaatatctt taacaataga
     4801 aatccatggg gtggggacaa gccgtaaaga ctggatgtcc atctcacacg aatttatggc
     4861 tatgggcaac acataatcct agtgcaatat gatactgggg ttattaagat gtgtcccagg
     4921 cagggaccaa gacaggtgaa ccatgttgtt acactctatt tgtaacaagg ggaaagagag
     4981 tggacgccga cagcagcgga ctccactggt tgtctctaac acccccgaaa attaaacggg
     5041 gctccacgcc aatggggccc ataaacaaag acaagtggcc actctttttt ttgaaattgt
     5101 ggagtggggg cacgcgtcag cccccacacg ccgccctgcg gttttggact gtaaaataag
     5161 ggtgtaataa cttggctgat tgtaaccccg ctaaccactg cggtcaaacc acttgcccac
     5221 aaaaccacta atggcacccc ggggaatacc tgcataagta ggtgggcggg ccaagatagg
     5281 ggcgcgattg ctgcgatctg gaggacaaat tacacacact tgcgcctgag cgccaagcac
     5341 agggttgttg gtcctcatat tcacgaggtc gctgagagca cggtgggcta atgttgccat
     5401 gggtagcata tactacccaa atatctggat agcatatgct atcctaatct atatctgggt
     5461 agcataggct atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt
     5521 agtatatgct atcctaattt atatctgggt agcataggct atcctaatct atatctgggt
     5581 agcatatgct atcctaatct atatctgggt agtatatgct atcctaatct gtatccgggt
     5641 agcatatgct atcctaatag agattagggt agtatatgct atcctaattt atatctgggt
     5701 agcatatact acccaaatat ctggatagca tatgctatcc taatctatat ctgggtagca
     5761 tatgctatcc taatctatat ctgggtagca taggctatcc taatctatat ctgggtagca
     5821 tatgctatcc taatctatat ctgggtagta tatgctatcc taatttatat ctgggtagca
     5881 taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagta
     5941 tatgctatcc taatctgtat ccgggtagca tatgctatcc tcatgataag ctgtcaaaca
     6001 tgagaatttt cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     6061 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     6121 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     6181 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     6241 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     6301 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     6361 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     6421 acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa
     6481 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     6541 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     6601 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     6661 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     6721 gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg
     6781 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     6841 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     6901 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     6961 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     7021 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     7081 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     7141 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     7201 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     7261 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     7321 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     7381 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     7441 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     7501 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     7561 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     7621 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     7681 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     7741 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     7801 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     7861 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     7921 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     7981 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     8041 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     8101 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     8161 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     8221 acaggaaaca gctatgacca tgattacgcc aagctctagc tagaggtcga ccaattctca
     8281 tgtttgacag cttatcatcg cagatccggg caacgttgtt gccattgctg caggcgcaga
     8341 actggtaggt atggaagatc tatacattga atcaatattg gcaattagcc atattagtca
     8401 ttggttatat agcataaatc aatattggct attggccatt gcatacgttg tatctatatc
     8461 ataatatgta catttatatt ggctcatgtc caatatgacc gc
//
