LOCUS       sequence-274540-        6900 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 6900)
  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 6900)
  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: D27_Ld_240060
FEATURES             Location/Qualifiers
     source          1..6900
                     /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             1678..1695
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1747..1802
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1801..1820)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4038..4056)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4124..4228
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4229..4297,4298..5089)
                     /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(4447..4466)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5260..5848
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5749..5768
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6002..6019
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6136..6157
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6172..6177,6178..6195,6196..6202)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6210..6226
                     /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     6215..6237
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6234..6250
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6234..6250
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        6521..6900
                     /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 gccaccgaca gcagcttcaa cttcgaggcc
      841 caggtcatcc tggaaaacgc caccgtgaac agcgacttcg tggtcaccac cggcttcacc
      901 gagctgatca ccacagccct gcagaacgtg gccaaccccc ccagccctct gagaaccgtg
      961 gtgcaggccg tgccccccca cctggtcatc atgctgagca tgtggtacac cggcgagccc
     1021 gagagcgtgc agaccgccat cagcaacatc aacaagacct acaccaagtg ggccagcggc
     1081 gacaccctgg ccgtgtctgg cgccctgagc cacacctgta gcaccaacat cggccagcag
     1141 cccaccttca tcaattacac caacagctgc agcagctggg acgccctgct gccccctagc
     1201 agcaatcaga cctgctggcg gaacctgagc ctgaccttct acaccgccgt gaccagccag
     1261 agcgacccca gagccagcct gcagtccagc ctgtgcagct tcctgcccac cgactgcggc
     1321 ctgatcacat acgacgacct gacccggaca caggtcacaa tcagcgacag cgtcgtgacc
     1381 gtgaatgtga tgcccttcac cgtgatgagc gagaacagag aggccacact ggccaccctg
     1441 gtcacctacg cccagtacgc cagcgtgctg gtggaacaca acatcgtgta catcctggcc
     1501 aacggcgtcc aggtgttctt tcagggcgac ccccagtcct tcagcatcaa cggcaccttc
     1561 gagcagtgcg tgaagaagat gggcgcgccc ggcagcctgc accacatcct ggacgcccag
     1621 aagatgctgt ggaaccacag ggacaggaac ctgcctcctc tggcccctct gggcccccac
     1681 caccatcacc accattgagt ctagacacgt gattaattaa ggatcccccg acctcgacct
     1741 ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct
     1801 cactcggaag gacatatggg agggcaaatc atttggtcga gatccctcgg agatctctag
     1861 ctagaggatc gatccccgcc ccggacgaac taaacctgac tacgacatct ctgccccttc
     1921 ttcgcggggc agtgcatgta atcccttcag ttggttggta caacttgcca actgggccct
     1981 gttccacatg tgacacgggg ggggaccaaa cacaaagggg ttctctgact gtagttgaca
     2041 tccttataaa tggatgtgca catttgccaa cactgagtgg ctttcatcct ggagcagact
     2101 ttgcagtctg tggactgcaa cacaacattg cctttatgtg taactcttgg ctgaagctct
     2161 tacaccaatg ctgggggaca tgtacctccc aggggcccag gaagactacg ggaggctaca
     2221 ccaacgtcaa tcagaggggc ctgtgtagct accgataagc ggaccctcaa gagggcatta
     2281 gcaatagtgt ttataaggcc cccttgttaa ccctaaacgg gtagcatatg cttcccgggt
     2341 agtagtatat actatccaga ctaaccctaa ttcaatagca tatgttaccc aacgggaagc
     2401 atatgctatc gaattagggt tagtaaaagg gtcctaagga acagcgatat ctcccacccc
     2461 atgagctgtc acggttttat ttacatgggg tcaggattcc acgagggtag tgaaccattt
     2521 tagtcacaag ggcagtggct gaagatcaag gagcgggcag tgaactctcc tgaatcttcg
     2581 cctgcttctt cattctcctt cgtttagcta atagaataac tgctgagttg tgaacagtaa
     2641 ggtgtatgtg aggtgctcga aaacaaggtt tcaggtgacg cccccagaat aaaatttgga
     2701 cggggggttc agtggtggca ttgtgctatg acaccaatat aaccctcaca aaccccttgg
     2761 gcaataaata ctagtgtagg aatgaaacat tctgaatatc tttaacaata gaaatccatg
     2821 gggtggggac aagccgtaaa gactggatgt ccatctcaca cgaatttatg gctatgggca
     2881 acacataatc ctagtgcaat atgatactgg ggttattaag atgtgtccca ggcagggacc
     2941 aagacaggtg aaccatgttg ttacactcta tttgtaacaa ggggaaagag agtggacgcc
     3001 gacagcagcg gactccactg gttgtctcta acacccccga aaattaaacg gggctccacg
     3061 ccaatggggc ccataaacaa agacaagtgg ccactctttt ttttgaaatt gtggagtggg
     3121 ggcacgcgtc agcccccaca cgccgccctg cggttttgga ctgtaaaata agggtgtaat
     3181 aacttggctg attgtaaccc cgctaaccac tgcggtcaaa ccacttgccc acaaaaccac
     3241 taatggcacc ccggggaata cctgcataag taggtgggcg ggccaagata ggggcgcgat
     3301 tgctgcgatc tggaggacaa attacacaca cttgcgcctg agcgccaagc acagggttgt
     3361 tggtcctcat attcacgagg tcgctgagag cacggtgggc taatgttgcc atgggtagca
     3421 tatactaccc aaatatctgg atagcatatg ctatcctaat ctatatctgg gtagcatagg
     3481 ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagtatatg
     3541 ctatcctaat ttatatctgg gtagcatagg ctatcctaat ctatatctgg gtagcatatg
     3601 ctatcctaat ctatatctgg gtagtatatg ctatcctaat ctgtatccgg gtagcatatg
     3661 ctatcctaat agagattagg gtagtatatg ctatcctaat ttatatctgg gtagcatata
     3721 ctacccaaat atctggatag catatgctat cctaatctat atctgggtag catatgctat
     3781 cctaatctat atctgggtag cataggctat cctaatctat atctgggtag catatgctat
     3841 cctaatctat atctgggtag tatatgctat cctaatttat atctgggtag cataggctat
     3901 cctaatctat atctgggtag catatgctat cctaatctat atctgggtag tatatgctat
     3961 cctaatctgt atccgggtag catatgctat cctcatgata agctgtcaaa catgagaatt
     4021 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     4081 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     4141 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4201 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     4261 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     4321 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     4381 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     4441 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
     4501 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     4561 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     4621 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     4681 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     4741 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact
     4801 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     4861 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     4921 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     4981 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     5041 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     5101 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     5161 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     5221 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     5281 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     5341 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     5401 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     5461 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     5521 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     5581 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     5641 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     5701 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     5761 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     5821 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     5881 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     5941 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     6001 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     6061 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     6121 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     6181 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     6241 cagctatgac catgattacg ccaagctcta gctagaggtc gaccaattct catgtttgac
     6301 agcttatcat cgcagatccg ggcaacgttg ttgccattgc tgcaggcgca gaactggtag
     6361 gtatggaaga tctatacatt gaatcaatat tggcaattag ccatattagt cattggttat
     6421 atagcataaa tcaatattgg ctattggcca ttgcatacgt tgtatctata tcataatatg
     6481 tacatttata ttggctcatg tccaatatga ccgccatgtt gacattgatt attgactagt
     6541 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     6601 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     6661 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     6721 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     6781 ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     6841 accttacggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
//
