LOCUS       sequence-274530-        6480 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 6480)
  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 6480)
  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: D15_Ld_061240
FEATURES             Location/Qualifiers
     source          1..6480
                     /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             1258..1275
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1327..1382
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1381..1400)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3618..3636)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3704..3808
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3809..3877,3878..4669)
                     /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(4027..4046)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4840..5428
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5329..5348
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5582..5599
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5716..5737
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5752..5757,5758..5775,5776..5782)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5790..5806
                     /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     5795..5817
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5814..5830
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5814..5830
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        6101..6480
                     /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 gccaccgccg aagcctctag agtggtggaa
      841 ctgacccggg acaccttcga cgcctacgtg ggcggacccg acacctacac ctttgtgctg
      901 tactgcgtga cctggtcccg gcagtgcaag accgccatcc agctgtggga ccggctgagc
      961 atcagccaga gccagaaaga actgcgggac gtgttcaccg ccgcctatgt ggacggcgac
     1021 aagtaccccg acctcgtgaa gcggatgaac gtgaagggct tccccaccat gctgttctac
     1081 acccccctgc acaccgaggg cctcgagtac aacggcccca gagagagcta cctgctggac
     1141 agcttcgtgt tccagttctc tggcgcgccc ggcagcctgc accacatcct ggacgcccag
     1201 aagatgctgt ggaaccacag ggacaggaac ctgcctcctc tggcccctct gggcccccac
     1261 caccatcacc accattgagt ctagacacgt gattaattaa ggatcccccg acctcgacct
     1321 ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct
     1381 cactcggaag gacatatggg agggcaaatc atttggtcga gatccctcgg agatctctag
     1441 ctagaggatc gatccccgcc ccggacgaac taaacctgac tacgacatct ctgccccttc
     1501 ttcgcggggc agtgcatgta atcccttcag ttggttggta caacttgcca actgggccct
     1561 gttccacatg tgacacgggg ggggaccaaa cacaaagggg ttctctgact gtagttgaca
     1621 tccttataaa tggatgtgca catttgccaa cactgagtgg ctttcatcct ggagcagact
     1681 ttgcagtctg tggactgcaa cacaacattg cctttatgtg taactcttgg ctgaagctct
     1741 tacaccaatg ctgggggaca tgtacctccc aggggcccag gaagactacg ggaggctaca
     1801 ccaacgtcaa tcagaggggc ctgtgtagct accgataagc ggaccctcaa gagggcatta
     1861 gcaatagtgt ttataaggcc cccttgttaa ccctaaacgg gtagcatatg cttcccgggt
     1921 agtagtatat actatccaga ctaaccctaa ttcaatagca tatgttaccc aacgggaagc
     1981 atatgctatc gaattagggt tagtaaaagg gtcctaagga acagcgatat ctcccacccc
     2041 atgagctgtc acggttttat ttacatgggg tcaggattcc acgagggtag tgaaccattt
     2101 tagtcacaag ggcagtggct gaagatcaag gagcgggcag tgaactctcc tgaatcttcg
     2161 cctgcttctt cattctcctt cgtttagcta atagaataac tgctgagttg tgaacagtaa
     2221 ggtgtatgtg aggtgctcga aaacaaggtt tcaggtgacg cccccagaat aaaatttgga
     2281 cggggggttc agtggtggca ttgtgctatg acaccaatat aaccctcaca aaccccttgg
     2341 gcaataaata ctagtgtagg aatgaaacat tctgaatatc tttaacaata gaaatccatg
     2401 gggtggggac aagccgtaaa gactggatgt ccatctcaca cgaatttatg gctatgggca
     2461 acacataatc ctagtgcaat atgatactgg ggttattaag atgtgtccca ggcagggacc
     2521 aagacaggtg aaccatgttg ttacactcta tttgtaacaa ggggaaagag agtggacgcc
     2581 gacagcagcg gactccactg gttgtctcta acacccccga aaattaaacg gggctccacg
     2641 ccaatggggc ccataaacaa agacaagtgg ccactctttt ttttgaaatt gtggagtggg
     2701 ggcacgcgtc agcccccaca cgccgccctg cggttttgga ctgtaaaata agggtgtaat
     2761 aacttggctg attgtaaccc cgctaaccac tgcggtcaaa ccacttgccc acaaaaccac
     2821 taatggcacc ccggggaata cctgcataag taggtgggcg ggccaagata ggggcgcgat
     2881 tgctgcgatc tggaggacaa attacacaca cttgcgcctg agcgccaagc acagggttgt
     2941 tggtcctcat attcacgagg tcgctgagag cacggtgggc taatgttgcc atgggtagca
     3001 tatactaccc aaatatctgg atagcatatg ctatcctaat ctatatctgg gtagcatagg
     3061 ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagtatatg
     3121 ctatcctaat ttatatctgg gtagcatagg ctatcctaat ctatatctgg gtagcatatg
     3181 ctatcctaat ctatatctgg gtagtatatg ctatcctaat ctgtatccgg gtagcatatg
     3241 ctatcctaat agagattagg gtagtatatg ctatcctaat ttatatctgg gtagcatata
     3301 ctacccaaat atctggatag catatgctat cctaatctat atctgggtag catatgctat
     3361 cctaatctat atctgggtag cataggctat cctaatctat atctgggtag catatgctat
     3421 cctaatctat atctgggtag tatatgctat cctaatttat atctgggtag cataggctat
     3481 cctaatctat atctgggtag catatgctat cctaatctat atctgggtag tatatgctat
     3541 cctaatctgt atccgggtag catatgctat cctcatgata agctgtcaaa catgagaatt
     3601 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     3661 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     3721 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     3781 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     3841 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     3901 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     3961 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     4021 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
     4081 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     4141 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     4201 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     4261 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     4321 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact
     4381 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     4441 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     4501 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     4561 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     4621 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     4681 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     4741 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     4801 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     4861 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     4921 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     4981 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     5041 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     5101 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     5161 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     5221 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     5281 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     5341 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     5401 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     5461 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     5521 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     5581 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     5641 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     5701 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     5761 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     5821 cagctatgac catgattacg ccaagctcta gctagaggtc gaccaattct catgtttgac
     5881 agcttatcat cgcagatccg ggcaacgttg ttgccattgc tgcaggcgca gaactggtag
     5941 gtatggaaga tctatacatt gaatcaatat tggcaattag ccatattagt cattggttat
     6001 atagcataaa tcaatattgg ctattggcca ttgcatacgt tgtatctata tcataatatg
     6061 tacatttata ttggctcatg tccaatatga ccgccatgtt gacattgatt attgactagt
     6121 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     6181 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     6241 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     6301 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     6361 ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     6421 accttacggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
//
