LOCUS       sequence-274469-        6744 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 6744)
  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 6744)
  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: D10_Ld_292250
FEATURES             Location/Qualifiers
     source          1..6744
                     /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             1902..1919
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1971..2026
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2025..2044)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4262..4280)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4348..4452
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4453..4521,4522..5313)
                     /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(4671..4690)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5484..6072
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5973..5992
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6226..6243
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6360..6381
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6396..6401,6402..6419,6420..6426)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6434..6450
                     /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     6439..6461
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6458..6474
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6458..6474
                     /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 gccaccttct
     1201 tcgcctacgt ggaccagccc agccactggg tgtacgaccc tgtgcggctg cagcagatcg
     1261 cccagcagtc tatcgccacc gcccacgccg ccaatggcgg caaggccaca gccaagcagg
     1321 tcacagacga gacaatccgg ctgatcctgg aaaactaccc caagaccacc cggtacaccg
     1381 gccactggct gtggaacaat gccggcggag ccatgggcag catgaccgtg ctgcactgca
     1441 gcttcagcga gtacctgatc atcttcggca gcagcgtggg caccgagggc cacaccggca
     1501 gatacttttg ggccgacgac ttcttcaaca tcctcgtggg cgagcagtgg gccgctctgc
     1561 ccggcgtgga agagaaagag gtgtacagac ccggcgacca gcacatcctg cccagaggca
     1621 gcgccaagca gtaccggatg cccgaggaat gctgggccct ggaatacgcc agaggcaaca
     1681 tcgtgtccat gctgttcttc ggcttcgccg acatgttcag cagcaccctg gacgtggtca
     1741 ccatctacca gaccgtgatg gaaagcgccg gcaacatgat ccggaacacc ctgatgggca
     1801 agattggcgc gcccggcagc ctgcaccaca tcctggacgc ccagaagatg ctgtggaacc
     1861 acagggacag gaacctgcct cctctggccc ctctgggccc ccaccaccat caccaccatt
     1921 gagtctagac acgtgattaa ttaaggatcc cccgacctcg acctctggct aataaaggaa
     1981 atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata
     2041 tgggagggca aatcatttgg tcgagatccc tcggagatct ctagctagag gatcgatccc
     2101 cgccccggac gaactaaacc tgactacgac atctctgccc cttcttcgcg gggcagtgca
     2161 tgtaatccct tcagttggtt ggtacaactt gccaactggg ccctgttcca catgtgacac
     2221 ggggggggac caaacacaaa ggggttctct gactgtagtt gacatcctta taaatggatg
     2281 tgcacatttg ccaacactga gtggctttca tcctggagca gactttgcag tctgtggact
     2341 gcaacacaac attgccttta tgtgtaactc ttggctgaag ctcttacacc aatgctgggg
     2401 gacatgtacc tcccaggggc ccaggaagac tacgggaggc tacaccaacg tcaatcagag
     2461 gggcctgtgt agctaccgat aagcggaccc tcaagagggc attagcaata gtgtttataa
     2521 ggcccccttg ttaaccctaa acgggtagca tatgcttccc gggtagtagt atatactatc
     2581 cagactaacc ctaattcaat agcatatgtt acccaacggg aagcatatgc tatcgaatta
     2641 gggttagtaa aagggtccta aggaacagcg atatctccca ccccatgagc tgtcacggtt
     2701 ttatttacat ggggtcagga ttccacgagg gtagtgaacc attttagtca caagggcagt
     2761 ggctgaagat caaggagcgg gcagtgaact ctcctgaatc ttcgcctgct tcttcattct
     2821 ccttcgttta gctaatagaa taactgctga gttgtgaaca gtaaggtgta tgtgaggtgc
     2881 tcgaaaacaa ggtttcaggt gacgccccca gaataaaatt tggacggggg gttcagtggt
     2941 ggcattgtgc tatgacacca atataaccct cacaaacccc ttgggcaata aatactagtg
     3001 taggaatgaa acattctgaa tatctttaac aatagaaatc catggggtgg ggacaagccg
     3061 taaagactgg atgtccatct cacacgaatt tatggctatg ggcaacacat aatcctagtg
     3121 caatatgata ctggggttat taagatgtgt cccaggcagg gaccaagaca ggtgaaccat
     3181 gttgttacac tctatttgta acaaggggaa agagagtgga cgccgacagc agcggactcc
     3241 actggttgtc tctaacaccc ccgaaaatta aacggggctc cacgccaatg gggcccataa
     3301 acaaagacaa gtggccactc ttttttttga aattgtggag tgggggcacg cgtcagcccc
     3361 cacacgccgc cctgcggttt tggactgtaa aataagggtg taataacttg gctgattgta
     3421 accccgctaa ccactgcggt caaaccactt gcccacaaaa ccactaatgg caccccgggg
     3481 aatacctgca taagtaggtg ggcgggccaa gataggggcg cgattgctgc gatctggagg
     3541 acaaattaca cacacttgcg cctgagcgcc aagcacaggg ttgttggtcc tcatattcac
     3601 gaggtcgctg agagcacggt gggctaatgt tgccatgggt agcatatact acccaaatat
     3661 ctggatagca tatgctatcc taatctatat ctgggtagca taggctatcc taatctatat
     3721 ctgggtagca tatgctatcc taatctatat ctgggtagta tatgctatcc taatttatat
     3781 ctgggtagca taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat
     3841 ctgggtagta tatgctatcc taatctgtat ccgggtagca tatgctatcc taatagagat
     3901 tagggtagta tatgctatcc taatttatat ctgggtagca tatactaccc aaatatctgg
     3961 atagcatatg ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg
     4021 gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg
     4081 gtagtatatg ctatcctaat ttatatctgg gtagcatagg ctatcctaat ctatatctgg
     4141 gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat ctgtatccgg
     4201 gtagcatatg ctatcctcat gataagctgt caaacatgag aattttcttg aagacgaaag
     4261 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     4321 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     4381 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     4441 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     4501 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     4561 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     4621 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     4681 gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     4741 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     4801 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     4861 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     4921 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     4981 gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     5041 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     5101 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     5161 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     5221 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     5281 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     5341 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     5401 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     5461 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     5521 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     5581 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     5641 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     5701 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     5761 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     5821 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     5881 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     5941 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6001 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     6061 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     6121 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     6181 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     6241 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     6301 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     6361 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     6421 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     6481 tacgccaagc tctagctaga ggtcgaccaa ttctcatgtt tgacagctta tcatcgcaga
     6541 tccgggcaac gttgttgcca ttgctgcagg cgcagaactg gtaggtatgg aagatctata
     6601 cattgaatca atattggcaa ttagccatat tagtcattgg ttatatagca taaatcaata
     6661 ttggctattg gccattgcat acgttgtatc tatatcataa tatgtacatt tatattggct
     6721 catgtccaat atgaccgcca tgtt
//
