LOCUS       sequence-274514-        6834 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 6834)
  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 6834)
  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: D65_Ld_051210
FEATURES             Location/Qualifiers
     source          1..6834
                     /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             1992..2009
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2061..2116
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2115..2134)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4352..4370)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4438..4542
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4543..4611,4612..5403)
                     /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(4761..4780)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5574..6162
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6063..6082
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6316..6333
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6450..6471
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6486..6491,6492..6509,6510..6516)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6524..6540
                     /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     6529..6551
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6548..6564
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6548..6564
                     /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 gccgatacca
     1201 ttaccgtaaa cgcagcaaca caagcctggt tgaagatgtg gattaaatca attcccaact
     1261 tgcagatcat ttggctgaac cctaatatct gctccagagc cggcattgag tgcgtgtctg
     1321 cgacaaacag tataaacatc cgactcgacg gggtcacgag cacaggcttt aactttatag
     1381 gaacgcttcc agaagttgac tcttctattg atggtaatga gctgcagatt acgtccattt
     1441 ccgtcagggg taaagcgcgt tttacgggca caattccagc atcctggagt agaatcacgc
     1501 ggcttacgcg cctggatttc agtaggaccc ggatgagcgg gcgcattcct gatgagctgg
     1561 ggtccctggc tacactggta agcgtggatt tctccaatag ctacttttgt tatgggctcc
     1621 caaactggaa tgctagcagt atgcctatgc tcagccaggc tatctttaca aacaataaca
     1681 tgcggggacc gttcgcaagc tcttggtcca ccttcccagc atcactcaag ctggacatca
     1741 ccgggaacaa gctttgcgga tgtatgcctt catcatggga cagcaagtcc aacctcgttg
     1801 cggcggctaa ggctatggac gcaggaactg tatccaactg tttcaggtct tgtaacagcg
     1861 catctttgag ctactgcccg gcacccccta ccggcggcgc gcccggcagc ctgcaccaca
     1921 tcctggacgc ccagaagatg ctgtggaacc acagggacag gaacctgcct cctctggccc
     1981 ctctgggccc ccaccaccat caccaccatt gagtctagac acgtgattaa ttaaggatcc
     2041 cccgacctcg acctctggct aataaaggaa atttattttc attgcaatag tgtgttggaa
     2101 ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttgg tcgagatccc
     2161 tcggagatct ctagctagag gatcgatccc cgccccggac gaactaaacc tgactacgac
     2221 atctctgccc cttcttcgcg gggcagtgca tgtaatccct tcagttggtt ggtacaactt
     2281 gccaactggg ccctgttcca catgtgacac ggggggggac caaacacaaa ggggttctct
     2341 gactgtagtt gacatcctta taaatggatg tgcacatttg ccaacactga gtggctttca
     2401 tcctggagca gactttgcag tctgtggact gcaacacaac attgccttta tgtgtaactc
     2461 ttggctgaag ctcttacacc aatgctgggg gacatgtacc tcccaggggc ccaggaagac
     2521 tacgggaggc tacaccaacg tcaatcagag gggcctgtgt agctaccgat aagcggaccc
     2581 tcaagagggc attagcaata gtgtttataa ggcccccttg ttaaccctaa acgggtagca
     2641 tatgcttccc gggtagtagt atatactatc cagactaacc ctaattcaat agcatatgtt
     2701 acccaacggg aagcatatgc tatcgaatta gggttagtaa aagggtccta aggaacagcg
     2761 atatctccca ccccatgagc tgtcacggtt ttatttacat ggggtcagga ttccacgagg
     2821 gtagtgaacc attttagtca caagggcagt ggctgaagat caaggagcgg gcagtgaact
     2881 ctcctgaatc ttcgcctgct tcttcattct ccttcgttta gctaatagaa taactgctga
     2941 gttgtgaaca gtaaggtgta tgtgaggtgc tcgaaaacaa ggtttcaggt gacgccccca
     3001 gaataaaatt tggacggggg gttcagtggt ggcattgtgc tatgacacca atataaccct
     3061 cacaaacccc ttgggcaata aatactagtg taggaatgaa acattctgaa tatctttaac
     3121 aatagaaatc catggggtgg ggacaagccg taaagactgg atgtccatct cacacgaatt
     3181 tatggctatg ggcaacacat aatcctagtg caatatgata ctggggttat taagatgtgt
     3241 cccaggcagg gaccaagaca ggtgaaccat gttgttacac tctatttgta acaaggggaa
     3301 agagagtgga cgccgacagc agcggactcc actggttgtc tctaacaccc ccgaaaatta
     3361 aacggggctc cacgccaatg gggcccataa acaaagacaa gtggccactc ttttttttga
     3421 aattgtggag tgggggcacg cgtcagcccc cacacgccgc cctgcggttt tggactgtaa
     3481 aataagggtg taataacttg gctgattgta accccgctaa ccactgcggt caaaccactt
     3541 gcccacaaaa ccactaatgg caccccgggg aatacctgca taagtaggtg ggcgggccaa
     3601 gataggggcg cgattgctgc gatctggagg acaaattaca cacacttgcg cctgagcgcc
     3661 aagcacaggg ttgttggtcc tcatattcac gaggtcgctg agagcacggt gggctaatgt
     3721 tgccatgggt agcatatact acccaaatat ctggatagca tatgctatcc taatctatat
     3781 ctgggtagca taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat
     3841 ctgggtagta tatgctatcc taatttatat ctgggtagca taggctatcc taatctatat
     3901 ctgggtagca tatgctatcc taatctatat ctgggtagta tatgctatcc taatctgtat
     3961 ccgggtagca tatgctatcc taatagagat tagggtagta tatgctatcc taatttatat
     4021 ctgggtagca tatactaccc aaatatctgg atagcatatg ctatcctaat ctatatctgg
     4081 gtagcatatg ctatcctaat ctatatctgg gtagcatagg ctatcctaat ctatatctgg
     4141 gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat ttatatctgg
     4201 gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg
     4261 gtagtatatg ctatcctaat ctgtatccgg gtagcatatg ctatcctcat gataagctgt
     4321 caaacatgag aattttcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt
     4381 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     4441 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     4501 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     4561 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     4621 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     4681 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     4741 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa
     4801 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     4861 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     4921 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     4981 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     5041 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca
     5101 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     5161 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     5221 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     5281 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     5341 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     5401 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     5461 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     5521 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     5581 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     5641 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     5701 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     5761 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     5821 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     5881 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     5941 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     6001 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     6061 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6121 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     6181 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     6241 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     6301 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     6361 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     6421 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     6481 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     6541 tttcacacag gaaacagcta tgaccatgat tacgccaagc tctagctaga ggtcgaccaa
     6601 ttctcatgtt tgacagctta tcatcgcaga tccgggcaac gttgttgcca ttgctgcagg
     6661 cgcagaactg gtaggtatgg aagatctata cattgaatca atattggcaa ttagccatat
     6721 tagtcattgg ttatatagca taaatcaata ttggctattg gccattgcat acgttgtatc
     6781 tatatcataa tatgtacatt tatattggct catgtccaat atgaccgcca tgtt
//
