LOCUS       sequence-262303-        7752 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 7752)
  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 7752)
  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: D8_Ld_340400
FEATURES             Location/Qualifiers
     source          1..7752
                     /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             2916..2933
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2985..3040
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3039..3058)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5276..5294)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5362..5466
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5467..5535,5536..6327)
                     /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(5685..5704)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6498..7086
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6987..7006
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7240..7257
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7374..7395
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7410..7415,7416..7433,7434..7440)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7448..7464
                     /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     7453..7475
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7472..7488
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7472..7488
                     /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 ccctgtctgc cgtggatcct gatgccggcg ctgacgccaa cggcagatct agccctgccc
     1261 ctgcctctgc cgtggccggc gagcacgtga ccgtgttcat gtgcgccggc agcagcgact
     1321 tcagcggcgt gcacgagctg ggcggcctga tgaaggccat ggaagagaca caggtgctga
     1381 gcctgaagta ctgccagacc gcccacctgg gctacatcag acacgccgat ggacacgccg
     1441 tgcggagcct ggtggtcacc accggcatca actacatcag cagcaccctg tgcaccagaa
     1501 gcgtgctgaa gctgcggcgg cagcggggct acacctacga cgccatggtg ttcatcggca
     1561 ccagcggctt tagccccatg gtcggaggct gggaccccac caacacaagc agctacgccg
     1621 ccagcaaggc cagcgagcag gaagccatga tgaagggcga agccgccaca gccggcgtgg
     1681 gagctgctcc tcctgccctg ccactgaaaa ccctgcagca ggtccgagag gaacaggccg
     1741 aggaagccca gctgagacag gccgctgtgg acggcggcca gctgtttacc ctgcctaccg
     1801 ccagccagct gaacttcgag aacgcccgga ccggcgaaca ggtggactgg gacggctgca
     1861 gccctagagt gcccagcgcc atcacacccc tgagcctggg cagcgtgtgt gtgaccagca
     1921 ccgcctacct gatggaaacc ggctgctgca ccgagcgcgt gcggcacagc cagtgcagca
     1981 gaccccactg caccagcttc cgcagcgagc tgggctccgt gaccaaggtg ttcttcacca
     2041 gcgacgccct ggcccggaag atcgaggccg cctctcacca caaggcctgg cccgccatgc
     2101 ccgagatcgt gaagaaaggc cagatgcggt tctgggccac caacgaggcc gtggaacctg
     2161 tgggcgaggg cgccgataga cacagagccg cccctagcgg ccccagcttc gtgacatgtg
     2221 ccgaggccac cggcaacacc atcaacacag gcgccgagcg ggactacctg tgcagagagc
     2281 acaccaccca ggccctggac atggcccagt acagactggc cgagctgcag gacggcaagg
     2341 ccgctcctca ggaacccccc ctgaccaccc agaatgtggc ctgcgtgcgg gctatggaat
     2401 ccatgggctt tctgcacagc gtggccgagg acagcaccgc cagagacatc cctgtggccg
     2461 tcgtgcgggg agccgccaac tacgacatgt accccatgaa gaaagtgtac gtgccccctg
     2521 ccgtgtggcg gagcgctctg gtggctggca gactgaaggt gtcccccaga gtggccgctg
     2581 ccctgaaggc cgaagaggcc gctgctgcca atggcgaggg ggccctgaga gagagcgcca
     2641 ttgccgccta cacctggcac cagaatgtgc acttcatgag cgaggccgag caccaggcct
     2701 tcgtgaacgc cagcttccac tacgccgtgg aaaccgccac cttcgtgctg agcaactact
     2761 tcttcggcgg cagatgcttc gccagcgcca gcggagccag acccgccaga agcaccgaag
     2821 gcgcgcccgg cagcctgcac cacatcctgg acgcccagaa gatgctgtgg aaccacaggg
     2881 acaggaacct gcctcctctg gcccctctgg gcccccacca ccatcaccac cattgagtct
     2941 agacacgtga ttaattaagg atcccccgac ctcgacctct ggctaataaa ggaaatttat
     3001 tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag
     3061 ggcaaatcat ttggtcgaga tccctcggag atctctagct agaggatcga tccccgcccc
     3121 ggacgaacta aacctgacta cgacatctct gccccttctt cgcggggcag tgcatgtaat
     3181 cccttcagtt ggttggtaca acttgccaac tgggccctgt tccacatgtg acacgggggg
     3241 ggaccaaaca caaaggggtt ctctgactgt agttgacatc cttataaatg gatgtgcaca
     3301 tttgccaaca ctgagtggct ttcatcctgg agcagacttt gcagtctgtg gactgcaaca
     3361 caacattgcc tttatgtgta actcttggct gaagctctta caccaatgct gggggacatg
     3421 tacctcccag gggcccagga agactacggg aggctacacc aacgtcaatc agaggggcct
     3481 gtgtagctac cgataagcgg accctcaaga gggcattagc aatagtgttt ataaggcccc
     3541 cttgttaacc ctaaacgggt agcatatgct tcccgggtag tagtatatac tatccagact
     3601 aaccctaatt caatagcata tgttacccaa cgggaagcat atgctatcga attagggtta
     3661 gtaaaagggt cctaaggaac agcgatatct cccaccccat gagctgtcac ggttttattt
     3721 acatggggtc aggattccac gagggtagtg aaccatttta gtcacaaggg cagtggctga
     3781 agatcaagga gcgggcagtg aactctcctg aatcttcgcc tgcttcttca ttctccttcg
     3841 tttagctaat agaataactg ctgagttgtg aacagtaagg tgtatgtgag gtgctcgaaa
     3901 acaaggtttc aggtgacgcc cccagaataa aatttggacg gggggttcag tggtggcatt
     3961 gtgctatgac accaatataa ccctcacaaa ccccttgggc aataaatact agtgtaggaa
     4021 tgaaacattc tgaatatctt taacaataga aatccatggg gtggggacaa gccgtaaaga
     4081 ctggatgtcc atctcacacg aatttatggc tatgggcaac acataatcct agtgcaatat
     4141 gatactgggg ttattaagat gtgtcccagg cagggaccaa gacaggtgaa ccatgttgtt
     4201 acactctatt tgtaacaagg ggaaagagag tggacgccga cagcagcgga ctccactggt
     4261 tgtctctaac acccccgaaa attaaacggg gctccacgcc aatggggccc ataaacaaag
     4321 acaagtggcc actctttttt ttgaaattgt ggagtggggg cacgcgtcag cccccacacg
     4381 ccgccctgcg gttttggact gtaaaataag ggtgtaataa cttggctgat tgtaaccccg
     4441 ctaaccactg cggtcaaacc acttgcccac aaaaccacta atggcacccc ggggaatacc
     4501 tgcataagta ggtgggcggg ccaagatagg ggcgcgattg ctgcgatctg gaggacaaat
     4561 tacacacact tgcgcctgag cgccaagcac agggttgttg gtcctcatat tcacgaggtc
     4621 gctgagagca cggtgggcta atgttgccat gggtagcata tactacccaa atatctggat
     4681 agcatatgct atcctaatct atatctgggt agcataggct atcctaatct atatctgggt
     4741 agcatatgct atcctaatct atatctgggt agtatatgct atcctaattt atatctgggt
     4801 agcataggct atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt
     4861 agtatatgct atcctaatct gtatccgggt agcatatgct atcctaatag agattagggt
     4921 agtatatgct atcctaattt atatctgggt agcatatact acccaaatat ctggatagca
     4981 tatgctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagca
     5041 taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagta
     5101 tatgctatcc taatttatat ctgggtagca taggctatcc taatctatat ctgggtagca
     5161 tatgctatcc taatctatat ctgggtagta tatgctatcc taatctgtat ccgggtagca
     5221 tatgctatcc tcatgataag ctgtcaaaca tgagaatttt cttgaagacg aaagggcctc
     5281 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     5341 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     5401 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     5461 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     5521 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     5581 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     5641 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     5701 ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     5761 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     5821 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     5881 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     5941 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     6001 acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     6061 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     6121 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     6181 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     6241 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     6301 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     6361 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     6421 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     6481 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     6541 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     6601 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     6661 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     6721 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     6781 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     6841 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     6901 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     6961 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     7021 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     7081 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     7141 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     7201 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     7261 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     7321 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     7381 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     7441 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     7501 aagctctagc tagaggtcga ccaattctca tgtttgacag cttatcatcg cagatccggg
     7561 caacgttgtt gccattgctg caggcgcaga actggtaggt atggaagatc tatacattga
     7621 atcaatattg gcaattagcc atattagtca ttggttatat agcataaatc aatattggct
     7681 attggccatt gcatacgttg tatctatatc ataatatgta catttatatt ggctcatgtc
     7741 caatatgacc gc
//
