LOCUS       sequence-274720-        7248 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 7248)
  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 7248)
  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: D42_Ld_242310
FEATURES             Location/Qualifiers
     source          1..7248
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     1086..1105
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1128..1137
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2406..2423
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2475..2530
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2529..2548)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4766..4784)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4852..4956
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4957..5025,5026..5817)
                     /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(5175..5194)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5988..6576
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6477..6496
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6730..6747
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6864..6885
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6900..6905,6906..6923,6924..6930)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6938..6954
                     /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     6943..6965
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6962..6978
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6962..6978
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agatcctcac tctcttccgc atcgctgtct gcgagggcca gctgttgggc tcgcggttga
      661 ggacaaactc ttcgcggtct ttccagtact cttggatcgg aaacccgtcg gcctccgaac
      721 ggtactccgc caccgaggga cctgagcgag tccgcatcga ccggatcgga aaacctctcg
      781 agaaaggcgt ctaaccagtc acagtcgcaa ggtaggctga gcaccgtggc gggcggcagc
      841 gggtggcggt cggggttgtt tctggcggag gtgctgctga tgatgtaatt aaagtaggcg
      901 gtcttgagac ggcggatggt cgaggtgagg tgtggcaggc ttgagatcca gctgttgggg
      961 tgagtactcc ctctcaaaag cgggcattac ttctgcgcta agattgtcag tttccaaaaa
     1021 cgaggaggat ttgatattca cctggcccga tctggccata cacttgagtg acaatgacat
     1081 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caagtttgcc accatggagt
     1141 ttcagaccca ggtactcatg tccctgctgc tctgcatgtc tggtgcggcc gccaccatct
     1201 atgtgatccc tgccgccgac gccgtggccc agttctgtag cgaacccccc ctgtgcgccc
     1261 ggaaggaccg gaccatgaag cagcagctgc tctgcaccat cagaagcgcc cctctgaagc
     1321 ccgccagccc cgtgtacaga cagtacgtgg tgtcccggct gaagagatac gtggacggca
     1381 gcatccggtc ccagagcgtg gcccccagcg tgatcgagag agtgcggtac aagctggccc
     1441 acatgcacga gcccatgatc ctgcacttcg ccggcgacaa cggcgtgggc aagaccagac
     1501 tggccgagct gatcagcctg gccatcggcc agaagtgcgg cgacgccacc tgtagcatcg
     1561 gcgataccac cctggtgctg agcggcacca gctacgacgg catgacagtg gccgagttcc
     1621 ggaatgccgt ggtgcccgtg gtcgtgcgcc acgcccagag ataccccgac aatggcgtcg
     1681 tgatcttcaa cgagctgacc tccctggaac ccagcaaagt gcgggtgctg tcccccctgc
     1741 tgggcagagg cacaagcttc cccgagaacc ccggcgtgtc cattgcccca ctgctggtca
     1801 tcctgaccac cgacttcggc agagagggcc ggacccgggg caagagcctg ttcgagatgc
     1861 gggccttcat caccgacgag ttcgccgacc tgtacagcaa agaggccgcc tcccacgtcc
     1921 ggaccttccc attcctgccc atctccctga gcacagccgg cgatatcgtg cgagtggtcg
     1981 tcagagagat cggctgcagc gccccacagc ccctgtgtct ggccattagc gacagcgccg
     2041 tcgtgtggct ggtggaacgg accaagatcc tgctgcccgc cgagaacggc agagccgtgg
     2101 ctttcgagac aaagctgcag ctggaagctc tgctggaaca ggtcatggac aactgcaccc
     2161 acgagagcgg catcattgcc accgatgagc tgtacctgga tgtggacgcc acatgcccct
     2221 accggcggtg caccgtgttt ctggaaggcg acggcacact ggccatgacc tgtcagggca
     2281 ccggcaccca cagcagagtg cctcccggag gcgcgcccgg cagcctgcac cacatcctgg
     2341 acgcccagaa gatgctgtgg aaccacaggg acaggaacct gcctcctctg gcccctctgg
     2401 gcccccacca ccatcaccac cattgagtct agacacgtga ttaattaagg atcccccgac
     2461 ctcgacctct ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt
     2521 gtgtctctca ctcggaagga catatgggag ggcaaatcat ttggtcgaga tccctcggag
     2581 atctctagct agaggatcga tccccgcccc ggacgaacta aacctgacta cgacatctct
     2641 gccccttctt cgcggggcag tgcatgtaat cccttcagtt ggttggtaca acttgccaac
     2701 tgggccctgt tccacatgtg acacgggggg ggaccaaaca caaaggggtt ctctgactgt
     2761 agttgacatc cttataaatg gatgtgcaca tttgccaaca ctgagtggct ttcatcctgg
     2821 agcagacttt gcagtctgtg gactgcaaca caacattgcc tttatgtgta actcttggct
     2881 gaagctctta caccaatgct gggggacatg tacctcccag gggcccagga agactacggg
     2941 aggctacacc aacgtcaatc agaggggcct gtgtagctac cgataagcgg accctcaaga
     3001 gggcattagc aatagtgttt ataaggcccc cttgttaacc ctaaacgggt agcatatgct
     3061 tcccgggtag tagtatatac tatccagact aaccctaatt caatagcata tgttacccaa
     3121 cgggaagcat atgctatcga attagggtta gtaaaagggt cctaaggaac agcgatatct
     3181 cccaccccat gagctgtcac ggttttattt acatggggtc aggattccac gagggtagtg
     3241 aaccatttta gtcacaaggg cagtggctga agatcaagga gcgggcagtg aactctcctg
     3301 aatcttcgcc tgcttcttca ttctccttcg tttagctaat agaataactg ctgagttgtg
     3361 aacagtaagg tgtatgtgag gtgctcgaaa acaaggtttc aggtgacgcc cccagaataa
     3421 aatttggacg gggggttcag tggtggcatt gtgctatgac accaatataa ccctcacaaa
     3481 ccccttgggc aataaatact agtgtaggaa tgaaacattc tgaatatctt taacaataga
     3541 aatccatggg gtggggacaa gccgtaaaga ctggatgtcc atctcacacg aatttatggc
     3601 tatgggcaac acataatcct agtgcaatat gatactgggg ttattaagat gtgtcccagg
     3661 cagggaccaa gacaggtgaa ccatgttgtt acactctatt tgtaacaagg ggaaagagag
     3721 tggacgccga cagcagcgga ctccactggt tgtctctaac acccccgaaa attaaacggg
     3781 gctccacgcc aatggggccc ataaacaaag acaagtggcc actctttttt ttgaaattgt
     3841 ggagtggggg cacgcgtcag cccccacacg ccgccctgcg gttttggact gtaaaataag
     3901 ggtgtaataa cttggctgat tgtaaccccg ctaaccactg cggtcaaacc acttgcccac
     3961 aaaaccacta atggcacccc ggggaatacc tgcataagta ggtgggcggg ccaagatagg
     4021 ggcgcgattg ctgcgatctg gaggacaaat tacacacact tgcgcctgag cgccaagcac
     4081 agggttgttg gtcctcatat tcacgaggtc gctgagagca cggtgggcta atgttgccat
     4141 gggtagcata tactacccaa atatctggat agcatatgct atcctaatct atatctgggt
     4201 agcataggct atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt
     4261 agtatatgct atcctaattt atatctgggt agcataggct atcctaatct atatctgggt
     4321 agcatatgct atcctaatct atatctgggt agtatatgct atcctaatct gtatccgggt
     4381 agcatatgct atcctaatag agattagggt agtatatgct atcctaattt atatctgggt
     4441 agcatatact acccaaatat ctggatagca tatgctatcc taatctatat ctgggtagca
     4501 tatgctatcc taatctatat ctgggtagca taggctatcc taatctatat ctgggtagca
     4561 tatgctatcc taatctatat ctgggtagta tatgctatcc taatttatat ctgggtagca
     4621 taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagta
     4681 tatgctatcc taatctgtat ccgggtagca tatgctatcc tcatgataag ctgtcaaaca
     4741 tgagaatttt cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     4801 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     4861 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4921 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     4981 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     5041 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     5101 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     5161 acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa
     5221 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     5281 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     5341 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     5401 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     5461 gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg
     5521 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     5581 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     5641 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     5701 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     5761 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     5821 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     5881 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5941 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     6001 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     6061 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     6121 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     6181 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     6241 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     6301 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     6361 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     6421 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     6481 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     6541 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     6601 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     6661 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     6721 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     6781 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     6841 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     6901 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     6961 acaggaaaca gctatgacca tgattacgcc aagctctagc tagaggtcga ccaattctca
     7021 tgtttgacag cttatcatcg cagatccggg caacgttgtt gccattgctg caggcgcaga
     7081 actggtaggt atggaagatc tatacattga atcaatattg gcaattagcc atattagtca
     7141 ttggttatat agcataaatc aatattggct attggccatt gcatacgttg tatctatatc
     7201 ataatatgta catttatatt ggctcatgtc caatatgacc gccatgtt
//
