LOCUS       sequence-262335-        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        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             2412..2429
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2481..2536
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2535..2554)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4772..4790)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4858..4962
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4963..5031,5032..5823)
                     /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(5181..5200)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5994..6582
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6483..6502
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6736..6753
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6870..6891
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6906..6911,6912..6929,6930..6936)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6944..6960
                     /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     6949..6971
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6968..6984
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6968..6984
                     /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 ccatctatgt gatccctgcc gccgacgccg tggcccagtt ctgtagcgaa ccccccctgt
     1261 gcgcccggaa ggaccggacc atgaagcagc agctgctctg caccatcaga agcgcccctc
     1321 tgaagcccgc cagccccgtg tacagacagt acgtggtgtc ccggctgaag agatacgtgg
     1381 acggcagcat ccggtcccag agcgtggccc ccagcgtgat cgagagagtg cggtacaagc
     1441 tggcccacat gcacgagccc atgatcctgc acttcgccgg cgacaacggc gtgggcaaga
     1501 ccagactggc cgagctgatc agcctggcca tcggccagaa gtgcggcgac gccacctgta
     1561 gcatcggcga taccaccctg gtgctgagcg gcaccagcta cgacggcatg acagtggccg
     1621 agttccggaa tgccgtggtg cccgtggtcg tgcgccacgc ccagagatac cccgacaatg
     1681 gcgtcgtgat cttcaacgag ctgacctccc tggaacccag caaagtgcgg gtgctgtccc
     1741 ccctgctggg cagaggcaca agcttccccg agaaccccgg cgtgtccatt gccccactgc
     1801 tggtcatcct gaccaccgac ttcggcagag agggccggac ccggggcaag agcctgttcg
     1861 agatgcgggc cttcatcacc gacgagttcg ccgacctgta cagcaaagag gccgcctccc
     1921 acgtccggac cttcccattc ctgcccatct ccctgagcac agccggcgat atcgtgcgag
     1981 tggtcgtcag agagatcggc tgcagcgccc cacagcccct gtgtctggcc attagcgaca
     2041 gcgccgtcgt gtggctggtg gaacggacca agatcctgct gcccgccgag aacggcagag
     2101 ccgtggcttt cgagacaaag ctgcagctgg aagctctgct ggaacaggtc atggacaact
     2161 gcacccacga gagcggcatc attgccaccg atgagctgta cctggatgtg gacgccacat
     2221 gcccctaccg gcggtgcacc gtgtttctgg aaggcgacgg cacactggcc atgacctgtc
     2281 agggcaccgg cacccacagc agagtgcctc ccggaggcgc gcccggcagc ctgcaccaca
     2341 tcctggacgc ccagaagatg ctgtggaacc acagggacag gaacctgcct cctctggccc
     2401 ctctgggccc ccaccaccat caccaccatt gagtctagac acgtgattaa ttaaggatcc
     2461 cccgacctcg acctctggct aataaaggaa atttattttc attgcaatag tgtgttggaa
     2521 ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttgg tcgagatccc
     2581 tcggagatct ctagctagag gatcgatccc cgccccggac gaactaaacc tgactacgac
     2641 atctctgccc cttcttcgcg gggcagtgca tgtaatccct tcagttggtt ggtacaactt
     2701 gccaactggg ccctgttcca catgtgacac ggggggggac caaacacaaa ggggttctct
     2761 gactgtagtt gacatcctta taaatggatg tgcacatttg ccaacactga gtggctttca
     2821 tcctggagca gactttgcag tctgtggact gcaacacaac attgccttta tgtgtaactc
     2881 ttggctgaag ctcttacacc aatgctgggg gacatgtacc tcccaggggc ccaggaagac
     2941 tacgggaggc tacaccaacg tcaatcagag gggcctgtgt agctaccgat aagcggaccc
     3001 tcaagagggc attagcaata gtgtttataa ggcccccttg ttaaccctaa acgggtagca
     3061 tatgcttccc gggtagtagt atatactatc cagactaacc ctaattcaat agcatatgtt
     3121 acccaacggg aagcatatgc tatcgaatta gggttagtaa aagggtccta aggaacagcg
     3181 atatctccca ccccatgagc tgtcacggtt ttatttacat ggggtcagga ttccacgagg
     3241 gtagtgaacc attttagtca caagggcagt ggctgaagat caaggagcgg gcagtgaact
     3301 ctcctgaatc ttcgcctgct tcttcattct ccttcgttta gctaatagaa taactgctga
     3361 gttgtgaaca gtaaggtgta tgtgaggtgc tcgaaaacaa ggtttcaggt gacgccccca
     3421 gaataaaatt tggacggggg gttcagtggt ggcattgtgc tatgacacca atataaccct
     3481 cacaaacccc ttgggcaata aatactagtg taggaatgaa acattctgaa tatctttaac
     3541 aatagaaatc catggggtgg ggacaagccg taaagactgg atgtccatct cacacgaatt
     3601 tatggctatg ggcaacacat aatcctagtg caatatgata ctggggttat taagatgtgt
     3661 cccaggcagg gaccaagaca ggtgaaccat gttgttacac tctatttgta acaaggggaa
     3721 agagagtgga cgccgacagc agcggactcc actggttgtc tctaacaccc ccgaaaatta
     3781 aacggggctc cacgccaatg gggcccataa acaaagacaa gtggccactc ttttttttga
     3841 aattgtggag tgggggcacg cgtcagcccc cacacgccgc cctgcggttt tggactgtaa
     3901 aataagggtg taataacttg gctgattgta accccgctaa ccactgcggt caaaccactt
     3961 gcccacaaaa ccactaatgg caccccgggg aatacctgca taagtaggtg ggcgggccaa
     4021 gataggggcg cgattgctgc gatctggagg acaaattaca cacacttgcg cctgagcgcc
     4081 aagcacaggg ttgttggtcc tcatattcac gaggtcgctg agagcacggt gggctaatgt
     4141 tgccatgggt agcatatact acccaaatat ctggatagca tatgctatcc taatctatat
     4201 ctgggtagca taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat
     4261 ctgggtagta tatgctatcc taatttatat ctgggtagca taggctatcc taatctatat
     4321 ctgggtagca tatgctatcc taatctatat ctgggtagta tatgctatcc taatctgtat
     4381 ccgggtagca tatgctatcc taatagagat tagggtagta tatgctatcc taatttatat
     4441 ctgggtagca tatactaccc aaatatctgg atagcatatg ctatcctaat ctatatctgg
     4501 gtagcatatg ctatcctaat ctatatctgg gtagcatagg ctatcctaat ctatatctgg
     4561 gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat ttatatctgg
     4621 gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg
     4681 gtagtatatg ctatcctaat ctgtatccgg gtagcatatg ctatcctcat gataagctgt
     4741 caaacatgag aattttcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt
     4801 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     4861 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     4921 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     4981 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     5041 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     5101 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     5161 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa
     5221 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     5281 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     5341 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     5401 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     5461 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca
     5521 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     5581 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     5641 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     5701 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     5761 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     5821 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     5881 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     5941 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     6001 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     6061 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     6121 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     6181 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     6241 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     6301 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     6361 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     6421 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     6481 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6541 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     6601 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     6661 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     6721 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     6781 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     6841 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     6901 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     6961 tttcacacag gaaacagcta tgaccatgat tacgccaagc tctagctaga ggtcgaccaa
     7021 ttctcatgtt tgacagctta tcatcgcaga tccgggcaac gttgttgcca ttgctgcagg
     7081 cgcagaactg gtaggtatgg aagatctata cattgaatca atattggcaa ttagccatat
     7141 tagtcattgg ttatatagca taaatcaata ttggctattg gccattgcat acgttgtatc
     7201 tatatcataa tatgtacatt tatattggct catgtccaat atgaccgc
//
