LOCUS       sequence-262327-        8232 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 8232)
  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 8232)
  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: D32_Ld_354780
FEATURES             Location/Qualifiers
     source          1..8232
                     /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             3396..3413
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3465..3520
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3519..3538)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5756..5774)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5842..5946
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5947..6015,6016..6807)
                     /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(6165..6184)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6978..7566
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7467..7486
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7720..7737
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7854..7875
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7890..7895,7896..7913,7914..7920)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7928..7944
                     /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     7933..7955
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7952..7968
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7952..7968
                     /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 cccacgtgat cggagtggat ctgggcagcg agtacatcaa ggtggccgga ccccacggcg
     1261 acaagggcgt ggacatcgtg ctgaacgagc agagcagaag aaagaccgac aacttcatcg
     1321 gcttccggcg gagcgacctg tacatcggcg acaccgccaa gagcctggcc gccagattcc
     1381 ctctgtgtac cgccagcgcc gtgaaccagc tcgtgcggat ccggaaggac agcagcctgc
     1441 tgccattctt ccgggacctg cagtacgagt accacgtgtc cttcaacaac cacggcagcg
     1501 ccaccgtgtc catctgcgat accaaggacc ccttcaccgc cgaggaactg tacagcatgg
     1561 tgctgagcta ctgcaagacc acagccgtga aggacgatgt ggtggacccc aagggggtgg
     1621 tggtcaccat ccccttcaga accagccctg ccgagcggag agccatcctg gacgccgcta
     1681 gactgagcgg cctgagcgtg ctgggcctga tgcacagcac caccgccgct gccttctact
     1741 acggcgtgcg gcaccggggc ttcggcaaca acaccctgag agtggtggtg ttcgacctgg
     1801 gctccaccca caccgaagtg ggcgtgtacg agttcctgcc tgccgcccag aaggcccccc
     1861 tgagcagcgc ctttggcaca ctgagaaccc tgggcgtggt ggaagatcgg agcctgggcg
     1921 gcagagcctt cgatctgtgc gtggccagag tgatcgaggc cgaggccaga gccaagctga
     1981 gcatcggccc tgtgctggga ggcaccacca gcgcccagct gaagtcccag ttctccctgc
     2041 tgagagccgc caacaaagtg cgggaaaccc tgtccgtgaa cagcaacacc ccctacaccg
     2101 tggaagggat cgcccccgac cgggactacc acagcagcat gaccagggcc accttcgaag
     2161 ccgagtgcgc cagcctgttc cagagagtga aggatctggc cacctccgtg accaaggtcg
     2221 tgaacatcag cttcaaggac ctgaacagct tcgagatgat gggcggcctg tccagaaccc
     2281 ccaagatcat tgccgacctg agcgaggcca tcggccggga cgtggaccgg accatgaaca
     2341 tggacgaggc tgccgccatt ggagccggct actacgccgc caaactgtcc cccctgtacc
     2401 gggccaagtc cctgaagctg gacgagacaa tcccctacgg ggtggacttc gaggtggaac
     2461 cccctctgtc caaagagaag cccgccgtca gacggcccct gttcggcgcc gatggcttcc
     2521 tgctgggcga cagcgtgtcc ctgaccttca accggaccga ggacttcagc ctgaacttct
     2581 acatcgatga cgatgccgcc aggcccttcg tgcgcgtgac cgtgacaggc gtgaagcagg
     2641 ccctgaccga catgaaggcc ctgtccccag ccgtggaaca tgccaacaac agccacatgg
     2701 tccgactgca gattgtgctg aatgagacag gcctggtgca ggtcgagcac gccgaggtgg
     2761 tcgtcagata cgccgaagaa gtgacccaga aagtcaccga gaacgtgacc gataaggcca
     2821 gcggcgagcg gaagtccgtg gaaaagtctg tgaaagtgat caagatgcgg atcagagccg
     2881 ctgacgtgac agccaccctg gcctggaaga accccaccgg cctgaccagc gaggaaatgg
     2941 aaaccagcca gacaaagctg aaaaccatct ggcaggccga acacgtgaaa cacctgaggg
     3001 ccactgccaa gaacaacctg gaagcctaca tcttctgggc caagaatgag ggcgtgtccg
     3061 acaacgccgc cctgacagct gctgctggcg ctgccgctgt gcagcgcgtg atcgatgaag
     3121 ccaccgccat gcaggaatgg ctggaagagg gcgagggcgc ctccgataat tgcgccgcca
     3181 gccagtatga gagcagactg gaaaacctgc ggggcatggt ggccgagctg accaagcagc
     3241 ccgaagaggc caccaagaca gtgaccaccg actccgccga cgacaacgag ggcgacctgg
     3301 gcgcgcccgg cagcctgcac cacatcctgg acgcccagaa gatgctgtgg aaccacaggg
     3361 acaggaacct gcctcctctg gcccctctgg gcccccacca ccatcaccac cattgagtct
     3421 agacacgtga ttaattaagg atcccccgac ctcgacctct ggctaataaa ggaaatttat
     3481 tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag
     3541 ggcaaatcat ttggtcgaga tccctcggag atctctagct agaggatcga tccccgcccc
     3601 ggacgaacta aacctgacta cgacatctct gccccttctt cgcggggcag tgcatgtaat
     3661 cccttcagtt ggttggtaca acttgccaac tgggccctgt tccacatgtg acacgggggg
     3721 ggaccaaaca caaaggggtt ctctgactgt agttgacatc cttataaatg gatgtgcaca
     3781 tttgccaaca ctgagtggct ttcatcctgg agcagacttt gcagtctgtg gactgcaaca
     3841 caacattgcc tttatgtgta actcttggct gaagctctta caccaatgct gggggacatg
     3901 tacctcccag gggcccagga agactacggg aggctacacc aacgtcaatc agaggggcct
     3961 gtgtagctac cgataagcgg accctcaaga gggcattagc aatagtgttt ataaggcccc
     4021 cttgttaacc ctaaacgggt agcatatgct tcccgggtag tagtatatac tatccagact
     4081 aaccctaatt caatagcata tgttacccaa cgggaagcat atgctatcga attagggtta
     4141 gtaaaagggt cctaaggaac agcgatatct cccaccccat gagctgtcac ggttttattt
     4201 acatggggtc aggattccac gagggtagtg aaccatttta gtcacaaggg cagtggctga
     4261 agatcaagga gcgggcagtg aactctcctg aatcttcgcc tgcttcttca ttctccttcg
     4321 tttagctaat agaataactg ctgagttgtg aacagtaagg tgtatgtgag gtgctcgaaa
     4381 acaaggtttc aggtgacgcc cccagaataa aatttggacg gggggttcag tggtggcatt
     4441 gtgctatgac accaatataa ccctcacaaa ccccttgggc aataaatact agtgtaggaa
     4501 tgaaacattc tgaatatctt taacaataga aatccatggg gtggggacaa gccgtaaaga
     4561 ctggatgtcc atctcacacg aatttatggc tatgggcaac acataatcct agtgcaatat
     4621 gatactgggg ttattaagat gtgtcccagg cagggaccaa gacaggtgaa ccatgttgtt
     4681 acactctatt tgtaacaagg ggaaagagag tggacgccga cagcagcgga ctccactggt
     4741 tgtctctaac acccccgaaa attaaacggg gctccacgcc aatggggccc ataaacaaag
     4801 acaagtggcc actctttttt ttgaaattgt ggagtggggg cacgcgtcag cccccacacg
     4861 ccgccctgcg gttttggact gtaaaataag ggtgtaataa cttggctgat tgtaaccccg
     4921 ctaaccactg cggtcaaacc acttgcccac aaaaccacta atggcacccc ggggaatacc
     4981 tgcataagta ggtgggcggg ccaagatagg ggcgcgattg ctgcgatctg gaggacaaat
     5041 tacacacact tgcgcctgag cgccaagcac agggttgttg gtcctcatat tcacgaggtc
     5101 gctgagagca cggtgggcta atgttgccat gggtagcata tactacccaa atatctggat
     5161 agcatatgct atcctaatct atatctgggt agcataggct atcctaatct atatctgggt
     5221 agcatatgct atcctaatct atatctgggt agtatatgct atcctaattt atatctgggt
     5281 agcataggct atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt
     5341 agtatatgct atcctaatct gtatccgggt agcatatgct atcctaatag agattagggt
     5401 agtatatgct atcctaattt atatctgggt agcatatact acccaaatat ctggatagca
     5461 tatgctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagca
     5521 taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagta
     5581 tatgctatcc taatttatat ctgggtagca taggctatcc taatctatat ctgggtagca
     5641 tatgctatcc taatctatat ctgggtagta tatgctatcc taatctgtat ccgggtagca
     5701 tatgctatcc tcatgataag ctgtcaaaca tgagaatttt cttgaagacg aaagggcctc
     5761 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     5821 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     5881 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     5941 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     6001 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     6061 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     6121 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     6181 ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     6241 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     6301 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     6361 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     6421 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     6481 acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     6541 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     6601 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     6661 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     6721 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     6781 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     6841 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     6901 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     6961 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     7021 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     7081 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     7141 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     7201 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     7261 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     7321 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     7381 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     7441 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     7501 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     7561 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     7621 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     7681 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     7741 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     7801 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     7861 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     7921 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     7981 aagctctagc tagaggtcga ccaattctca tgtttgacag cttatcatcg cagatccggg
     8041 caacgttgtt gccattgctg caggcgcaga actggtaggt atggaagatc tatacattga
     8101 atcaatattg gcaattagcc atattagtca ttggttatat agcataaatc aatattggct
     8161 attggccatt gcatacgttg tatctatatc ataatatgta catttatatt ggctcatgtc
     8221 caatatgacc gc
//
