LOCUS       sequence-274541-       10119 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 10119)
  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 10119)
  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: D25_Ld_190160
FEATURES             Location/Qualifiers
     source          1..10119
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     201..225
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     706..725
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      748..757
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4897..4914
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4966..5021
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5020..5039)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(7257..7275)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7343..7447
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7448..7516,7517..8308)
                     /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(7666..7685)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8479..9067
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8968..8987
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9221..9238
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9355..9376
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9391..9396,9397..9414,9415..9421)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9429..9445
                     /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     9434..9456
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9453..9469
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9453..9469
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        9740..10119
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta caccaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcctcac tctcttccgc
      241 atcgctgtct gcgagggcca gctgttgggc tcgcggttga ggacaaactc ttcgcggtct
      301 ttccagtact cttggatcgg aaacccgtcg gcctccgaac ggtactccgc caccgaggga
      361 cctgagcgag tccgcatcga ccggatcgga aaacctctcg agaaaggcgt ctaaccagtc
      421 acagtcgcaa ggtaggctga gcaccgtggc gggcggcagc gggtggcggt cggggttgtt
      481 tctggcggag gtgctgctga tgatgtaatt aaagtaggcg gtcttgagac ggcggatggt
      541 cgaggtgagg tgtggcaggc ttgagatcca gctgttgggg tgagtactcc ctctcaaaag
      601 cgggcattac ttctgcgcta agattgtcag tttccaaaaa cgaggaggat ttgatattca
      661 cctggcccga tctggccata cacttgagtg acaatgacat ccactttgcc tttctctcca
      721 caggtgtcca ctcccaggtc caagtttgcc accatggagt ttcagaccca ggtactcatg
      781 tccctgctgc tctgcatgtc tggtgcggcc gccaccgctc tgacagcccc ttttggcggc
      841 tgtgaagccg gcaccagcgg ctacgccatt cacagccctg gcgagaacga caagggcgcc
      901 cagtacctga tggacaacgg aatcgccttc gccatcctgc ctgccaccgt gaccaacggc
      961 gtggtcacct gggagaatgg cgaggtggcc gcctggatgc cccccgattt tcaggcccct
     1021 agcgacgccc actactgcct gatcctgcgg aagaacggcg cttggagcgt ggtgtccatg
     1081 cccttcacct ggaaccacct gtgcatcgac ggcagagagg acggcgagac aaccaccacc
     1141 gccacccaga tgcccaacac caccacaacc tctaccacca ctaccacccc tgcccctacc
     1201 gccaaccccc acctgagcag ctccagcttc cagagcgagg gcagcgccag cagcagcttc
     1261 gccagcccca tcacaacaac cacactggcc cccagcccta ccacctccac aacagccgcc
     1321 cctccaccta gcgtgcccac cgtgtctagc gtgatgcaga ccgtgcgctc cgtggtgttc
     1381 gccgccgtgc ctacagtgaa cgtgatggac ttcaccagca acgtggtgta cctgccagcc
     1441 acagccaccc tggtggaatc catgcaggcc ctgcaggccg ccagcatcgt gatcgccgtc
     1501 agatttggcg ccaccggccc tgtgtccgcc ctgctgacag atgccaccgc cgcctccgtg
     1561 tccttccacg tgccccacga gctgtacaac gtgtccgtcg tgaccagcca cgccctgagc
     1621 ctggtcaccc tgaacaccgc cgatgtgctg gtcgtgggca gccagctgag aatcaccccc
     1681 gtgctgatcg ctctgcagca gatgagcatg ggcgtggtgc tgagcgacga gagcatcctg
     1741 gaaggcaccg acatcctggc cacagtccgc tttgccggcg ctggcaccgc actgctgtac
     1801 accaccgacg tgtccaaagt gatcacaagc accctgagct tcagcatggt caacaacggc
     1861 gccgtgggct cccggtgcaa gcccagcaca agcagcatcc cttacttcat caccagatac
     1921 gacgtgcggt tcctggtgcc cggcaactac accgccagag tgcacgccgg cagaaagtcc
     1981 gtgtgcgcca cctacaacgg caagtctgtg gtggtggccc ccctggccat ccagggcatt
     2041 gccgccacaa ccactgccgc tagctttccc gccgctacag ccatcccaat ccggctgcag
     2101 ggcggcacaa ctaccggaat tgccagaggc gcctacgccg cctttgccag cgcctccagc
     2161 gattgcacct ccgagggcgc caagatcagc ctgctgcccg tgaacagctt caacagctcc
     2221 tccgacagct ctgccgtgcc aggcgctctg ctggtggcca gctctgaggt gccaaccggc
     2281 agctacgtgt gcatgcggcg gaccagcacc aataccaagt tcgccgtgcg gtatcacggc
     2341 aagcccgtga ccgtgggagc cgcctctatg cctcctgccg ctgctgccac tgcccctgtg
     2401 ctgaccatct tcaacggcag ctgcggctcc ctgaacctgc tggaatttgt gcggcctccc
     2461 gccagcgagg acgatatcgc cgcctacctg agcgccgctt ccaatgtgtc cgaggcctac
     2521 cgggtgttca gattcgtggc cgtgaaagcc ggcgcaactg ccgccgataa ggccgacgcc
     2581 tgccggcggt ttaagcagga agccaccagg gacgccgcca atggcgccat gcagttcagc
     2641 ttcccattct atctgcagca gaccaatggc gtgctgtgcg ccgacgagga atacgtgcgg
     2701 atcgattaca ccgtgatcca gcccaccaac tggctgatgt ccatggccgg cgtgtccgac
     2761 ttcgcctccc ggatgctggt gctgtccagc ggccctgagt acgacgctgt gcctctgagc
     2821 gtgcggagca gcaggctggc ctctgtgggc tctggcacag ccgtcaggat cagccaggac
     2881 ggctcctgcg ccttcggcta cacagcccgg atccagagct acaacaccca gcagtacatt
     2941 gccgtcccca ctgccagcgt gggcgtgtac acactgtgtg ccggcctgta caatgagaca
     3001 ggcacctcca tgttcgtgtc caccggctcc caggtggacc tggcctccta tgtgcggggc
     3061 agacggaagt ggatgcacga ggccggcatc gagagaaccg agtggacccc ctgtggctcc
     3121 gcctccacat gtggcctgca cacacagcag gcctacatcg aagtgtgcaa cggggctgtg
     3181 cggagaaacg tgtggcccca gagactggac ggcgcctttg tgggaagcga cagcgccatg
     3241 gtcctgatgg ccggaaatct gtacaagacc caggccattg tgctgatgaa cgacgtgtac
     3301 gtgcccgtgg ccgctgcctt cgctcactgt cagcagttct ccctgctgca gctgagcacc
     3361 gacgacggcg ctacctatgg cccagccgcc gtcgtgctgg gcaaccaccc tcagatcgcc
     3421 ctgtccccag tgactgatgt ctccggcgtg atcgtgggct tcgtgccctg gaacgccatg
     3481 tgcagaggcg tgcacacagg cgctgatgtg atggtggctc ggaccgccgc tggcatctcc
     3541 gtgatcgaca tcagcggcct gctgtttacc aagtctgagg gctactacgc cgtgtgtgtg
     3601 cggaacgggg tggcctgggc tagagtgacc gacacctaca tcaaggtggt catcggcggc
     3661 gtgacagtgt accaggccga ggccatcaac accggcgacg ctctggccaa ccagaagtcc
     3721 atccacctgt accagacaca gaccgccaca tggggcgtgg ccggcacatt cagcccttcc
     3781 gtgtctctgc tgctgaagct cgtgtggatg gacagcacat gcaccatgga tgccgcctat
     3841 accgctgccc tgcggtacat cagccctagc agcgcccatg ccacagtgga agccagcctg
     3901 attgctgccg ccccagctgg cgtggcatcc cccaccctgt acccctgcta catggccaac
     3961 accttcggca ataccgccgc acccatgcct ttcgaggaac acagaggccg gtccatcaca
     4021 gtgacccagc tgaccctgtc ctccctgttc tacctgccct atctggaact gtctgtgtct
     4081 gcccccgtgg acacccggat tctgctgacc gagccccagg acccccctat cacccgggtg
     4141 tccctgagag tgacaagctc ccagtctgat gtgccccagt gtggcgccgc agccacatac
     4201 gctacagtgc tgacaatcga gactgacgac ctgggcaggc ggtggatgac cgtgcctgcc
     4261 tgggtgccac agacactggg cggcagcggc gccacaaatg tgtatctgaa gatgtgcgca
     4321 accgcctcct ctagcgccaa tcccgagttt gtggccgtcg aagcctacct cgtgctgtct
     4381 gccaacgcca ccaacctcgc cggcaacccc atgagcctgc ggctgacctt cctgcctctc
     4441 aacagcagca ccgatctgct ggactacaag gccgctcagt atgccctcgt gaagtacgcc
     4501 gcaaccgagc tgaaggtgcc cgagacagtg gtgctcgtgg ctagcctggg aaacaacgcc
     4561 ttcgagctgt tcatcgatga cgccctccag accggcgcag cccctcagag cacactgacc
     4621 cctactcagc agctgtatca gaccctggct aaggggacca atctgcctct gttcctgaac
     4681 ggccgggata acgccacagc ttggtttgcc ctggcaaagg ctgaggccgt gcacctcgtg
     4741 gaccagacct ccgtggacga ttaccccgcc ctggccctgc cagtgtctgg atctgccgga
     4801 ggcgcgcccg gcagcctgca ccacatcctg gacgcccaga agatgctgtg gaaccacagg
     4861 gacaggaacc tgcctcctct ggcccctctg ggcccccacc accatcacca ccattgagtc
     4921 tagacacgtg attaattaag gatcccccga cctcgacctc tggctaataa aggaaattta
     4981 ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga
     5041 gggcaaatca tttggtcgag atccctcgga gatctctagc tagaggatcg atccccgccc
     5101 cggacgaact aaacctgact acgacatctc tgccccttct tcgcggggca gtgcatgtaa
     5161 tcccttcagt tggttggtac aacttgccaa ctgggccctg ttccacatgt gacacggggg
     5221 gggaccaaac acaaaggggt tctctgactg tagttgacat ccttataaat ggatgtgcac
     5281 atttgccaac actgagtggc tttcatcctg gagcagactt tgcagtctgt ggactgcaac
     5341 acaacattgc ctttatgtgt aactcttggc tgaagctctt acaccaatgc tgggggacat
     5401 gtacctccca ggggcccagg aagactacgg gaggctacac caacgtcaat cagaggggcc
     5461 tgtgtagcta ccgataagcg gaccctcaag agggcattag caatagtgtt tataaggccc
     5521 ccttgttaac cctaaacggg tagcatatgc ttcccgggta gtagtatata ctatccagac
     5581 taaccctaat tcaatagcat atgttaccca acgggaagca tatgctatcg aattagggtt
     5641 agtaaaaggg tcctaaggaa cagcgatatc tcccacccca tgagctgtca cggttttatt
     5701 tacatggggt caggattcca cgagggtagt gaaccatttt agtcacaagg gcagtggctg
     5761 aagatcaagg agcgggcagt gaactctcct gaatcttcgc ctgcttcttc attctccttc
     5821 gtttagctaa tagaataact gctgagttgt gaacagtaag gtgtatgtga ggtgctcgaa
     5881 aacaaggttt caggtgacgc ccccagaata aaatttggac ggggggttca gtggtggcat
     5941 tgtgctatga caccaatata accctcacaa accccttggg caataaatac tagtgtagga
     6001 atgaaacatt ctgaatatct ttaacaatag aaatccatgg ggtggggaca agccgtaaag
     6061 actggatgtc catctcacac gaatttatgg ctatgggcaa cacataatcc tagtgcaata
     6121 tgatactggg gttattaaga tgtgtcccag gcagggacca agacaggtga accatgttgt
     6181 tacactctat ttgtaacaag gggaaagaga gtggacgccg acagcagcgg actccactgg
     6241 ttgtctctaa cacccccgaa aattaaacgg ggctccacgc caatggggcc cataaacaaa
     6301 gacaagtggc cactcttttt tttgaaattg tggagtgggg gcacgcgtca gcccccacac
     6361 gccgccctgc ggttttggac tgtaaaataa gggtgtaata acttggctga ttgtaacccc
     6421 gctaaccact gcggtcaaac cacttgccca caaaaccact aatggcaccc cggggaatac
     6481 ctgcataagt aggtgggcgg gccaagatag gggcgcgatt gctgcgatct ggaggacaaa
     6541 ttacacacac ttgcgcctga gcgccaagca cagggttgtt ggtcctcata ttcacgaggt
     6601 cgctgagagc acggtgggct aatgttgcca tgggtagcat atactaccca aatatctgga
     6661 tagcatatgc tatcctaatc tatatctggg tagcataggc tatcctaatc tatatctggg
     6721 tagcatatgc tatcctaatc tatatctggg tagtatatgc tatcctaatt tatatctggg
     6781 tagcataggc tatcctaatc tatatctggg tagcatatgc tatcctaatc tatatctggg
     6841 tagtatatgc tatcctaatc tgtatccggg tagcatatgc tatcctaata gagattaggg
     6901 tagtatatgc tatcctaatt tatatctggg tagcatatac tacccaaata tctggatagc
     6961 atatgctatc ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagc
     7021 ataggctatc ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt
     7081 atatgctatc ctaatttata tctgggtagc ataggctatc ctaatctata tctgggtagc
     7141 atatgctatc ctaatctata tctgggtagt atatgctatc ctaatctgta tccgggtagc
     7201 atatgctatc ctcatgataa gctgtcaaac atgagaattt tcttgaagac gaaagggcct
     7261 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     7321 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     7381 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     7441 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     7501 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     7561 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     7621 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     7681 attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     7741 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     7801 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     7861 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     7921 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     7981 cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     8041 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     8101 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     8161 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     8221 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     8281 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     8341 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     8401 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     8461 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     8521 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     8581 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     8641 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     8701 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     8761 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     8821 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     8881 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     8941 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     9001 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     9061 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     9121 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     9181 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     9241 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     9301 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     9361 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     9421 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
     9481 caagctctag ctagaggtcg accaattctc atgtttgaca gcttatcatc gcagatccgg
     9541 gcaacgttgt tgccattgct gcaggcgcag aactggtagg tatggaagat ctatacattg
     9601 aatcaatatt ggcaattagc catattagtc attggttata tagcataaat caatattggc
     9661 tattggccat tgcatacgtt gtatctatat cataatatgt acatttatat tggctcatgt
     9721 ccaatatgac cgccatgttg acattgatta ttgactagtt attaatagta atcaattacg
     9781 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
     9841 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     9901 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     9961 gcccacttgg cagtacatca agtgtatcat atgccaagtc cgccccctat tgacgtcaat
    10021 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttacggga ctttcctact
    10081 tggcagtaca tctacgtatt agtcatcgct attaccatg
//
