LOCUS       sequence-262364-        7452 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 7452)
  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 7452)
  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: D72_Ld_231020
FEATURES             Location/Qualifiers
     source          1..7452
                     /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             2616..2633
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2685..2740
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2739..2758)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4976..4994)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5062..5166
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5167..5235,5236..6027)
                     /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(5385..5404)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6198..6786
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6687..6706
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6940..6957
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7074..7095
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7110..7115,7116..7133,7134..7140)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7148..7164
                     /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     7153..7175
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7172..7188
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7172..7188
                     /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 gcggccgccg
     1201 aagaagctga gatacaagcc aatgcatggt tgtccgcccc cgcatctgta tggagtgtta
     1261 tcgtctccga ccccagaact cacctcgcaa tgattcaggc tgttactgca gacttcgtgt
     1321 cagcgattag ggcggcagat agtaaggcag tggtgtttgt gacatccctg aagcctgtta
     1381 ctccccagaa agcctgggct tcaggccctg cctatggcgc ttacgctcag ttcacggcca
     1441 tgacccggct gagtaaagag gaagtgcttt gggcccttag aagttcagat ttgcgcacca
     1501 tcaataagat tttccagagt gtagggttcg acgcagctgt ggaagccctc gctgccgacg
     1561 gtccacccag tatgcaagct gttgggcctg tgctgctccc agtgagtgga tcaaccttct
     1621 tctggcagct cctgttggga acgctgatga ctcagaatgc aatcagcgta gatgccttca
     1681 tgtatcccct tcagatggac atccaggatg cagtggactc cacactgcga aactacttga
     1741 ttgaggctac acccctcacc aacgaggtga atgtatcctc atggtacgta tcctataacg
     1801 tgtgggcctt tcctgaggca gaggtcaccc cctctacccc tagtgacttg cgggccttcg
     1861 ccatgaagtc acgtatgcct ctctttaata gatatatgaa tacggtgatg gatacctatc
     1921 caacactgaa ggcttattca gacctgctcc cgaccatggt gcacgcgagg acgccacctc
     1981 tgggtaaccc attcgaaggc aatggtgcca atgaccctga caggatcttc gaggttttgg
     2041 ccaacgagac cggagattac cttcttgtgt tcagcggaga cgcggatcgt tggggacaag
     2101 tctgggaaaa gtcaagagac gctgtctcca aatccatcga ggaagctatc gaagacaggc
     2161 tgcacaggcg aacattcctc aatagagctt tcgtcacatc cgctaaaacc cttcccaacg
     2221 actctgtgaa cgtcggcggg ctgcaggttc tgtgccgcgt gctccagggc ataacaatga
     2281 tatccaacca cccttggagc cctgaagagg tgtccccact gttgctccag gccaattaca
     2341 gcgccaccca ggctctctac atctctggcg gtggacctgt tgcagagtac acgaagcgac
     2401 ccccagccac ggcttcccaa agcatcctgc ttgctgcttc atataaagcg agccctggct
     2461 ccattgcgac aacttccatc tcttatgact acagggtccg gctgaataag tatatcatag
     2521 gcgcgcccgg cagcctgcac cacatcctgg acgcccagaa gatgctgtgg aaccacaggg
     2581 acaggaacct gcctcctctg gcccctctgg gcccccacca ccatcaccac cattgagtct
     2641 agacacgtga ttaattaagg atcccccgac ctcgacctct ggctaataaa ggaaatttat
     2701 tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag
     2761 ggcaaatcat ttggtcgaga tccctcggag atctctagct agaggatcga tccccgcccc
     2821 ggacgaacta aacctgacta cgacatctct gccccttctt cgcggggcag tgcatgtaat
     2881 cccttcagtt ggttggtaca acttgccaac tgggccctgt tccacatgtg acacgggggg
     2941 ggaccaaaca caaaggggtt ctctgactgt agttgacatc cttataaatg gatgtgcaca
     3001 tttgccaaca ctgagtggct ttcatcctgg agcagacttt gcagtctgtg gactgcaaca
     3061 caacattgcc tttatgtgta actcttggct gaagctctta caccaatgct gggggacatg
     3121 tacctcccag gggcccagga agactacggg aggctacacc aacgtcaatc agaggggcct
     3181 gtgtagctac cgataagcgg accctcaaga gggcattagc aatagtgttt ataaggcccc
     3241 cttgttaacc ctaaacgggt agcatatgct tcccgggtag tagtatatac tatccagact
     3301 aaccctaatt caatagcata tgttacccaa cgggaagcat atgctatcga attagggtta
     3361 gtaaaagggt cctaaggaac agcgatatct cccaccccat gagctgtcac ggttttattt
     3421 acatggggtc aggattccac gagggtagtg aaccatttta gtcacaaggg cagtggctga
     3481 agatcaagga gcgggcagtg aactctcctg aatcttcgcc tgcttcttca ttctccttcg
     3541 tttagctaat agaataactg ctgagttgtg aacagtaagg tgtatgtgag gtgctcgaaa
     3601 acaaggtttc aggtgacgcc cccagaataa aatttggacg gggggttcag tggtggcatt
     3661 gtgctatgac accaatataa ccctcacaaa ccccttgggc aataaatact agtgtaggaa
     3721 tgaaacattc tgaatatctt taacaataga aatccatggg gtggggacaa gccgtaaaga
     3781 ctggatgtcc atctcacacg aatttatggc tatgggcaac acataatcct agtgcaatat
     3841 gatactgggg ttattaagat gtgtcccagg cagggaccaa gacaggtgaa ccatgttgtt
     3901 acactctatt tgtaacaagg ggaaagagag tggacgccga cagcagcgga ctccactggt
     3961 tgtctctaac acccccgaaa attaaacggg gctccacgcc aatggggccc ataaacaaag
     4021 acaagtggcc actctttttt ttgaaattgt ggagtggggg cacgcgtcag cccccacacg
     4081 ccgccctgcg gttttggact gtaaaataag ggtgtaataa cttggctgat tgtaaccccg
     4141 ctaaccactg cggtcaaacc acttgcccac aaaaccacta atggcacccc ggggaatacc
     4201 tgcataagta ggtgggcggg ccaagatagg ggcgcgattg ctgcgatctg gaggacaaat
     4261 tacacacact tgcgcctgag cgccaagcac agggttgttg gtcctcatat tcacgaggtc
     4321 gctgagagca cggtgggcta atgttgccat gggtagcata tactacccaa atatctggat
     4381 agcatatgct atcctaatct atatctgggt agcataggct atcctaatct atatctgggt
     4441 agcatatgct atcctaatct atatctgggt agtatatgct atcctaattt atatctgggt
     4501 agcataggct atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt
     4561 agtatatgct atcctaatct gtatccgggt agcatatgct atcctaatag agattagggt
     4621 agtatatgct atcctaattt atatctgggt agcatatact acccaaatat ctggatagca
     4681 tatgctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagca
     4741 taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagta
     4801 tatgctatcc taatttatat ctgggtagca taggctatcc taatctatat ctgggtagca
     4861 tatgctatcc taatctatat ctgggtagta tatgctatcc taatctgtat ccgggtagca
     4921 tatgctatcc tcatgataag ctgtcaaaca tgagaatttt cttgaagacg aaagggcctc
     4981 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     5041 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     5101 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     5161 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     5221 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     5281 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     5341 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     5401 ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     5461 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     5521 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     5581 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     5641 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     5701 acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     5761 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     5821 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     5881 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     5941 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     6001 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     6061 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     6121 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     6181 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     6241 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     6301 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     6361 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     6421 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     6481 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     6541 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     6601 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     6661 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     6721 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     6781 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     6841 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     6901 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     6961 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     7021 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     7081 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     7141 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     7201 aagctctagc tagaggtcga ccaattctca tgtttgacag cttatcatcg cagatccggg
     7261 caacgttgtt gccattgctg caggcgcaga actggtaggt atggaagatc tatacattga
     7321 atcaatattg gcaattagcc atattagtca ttggttatat agcataaatc aatattggct
     7381 attggccatt gcatacgttg tatctatatc ataatatgta catttatatt ggctcatgtc
     7441 caatatgacc gc
//
