LOCUS       sequence-274526-        7140 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 7140)
  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 7140)
  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: D11_Ld_211610
FEATURES             Location/Qualifiers
     source          1..7140
                     /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             1918..1935
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1987..2042
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2041..2060)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4278..4296)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4364..4468
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4469..4537,4538..5329)
                     /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(4687..4706)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5500..6088
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5989..6008
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6242..6259
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6376..6397
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6412..6417,6418..6435,6436..6442)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6450..6466
                     /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     6455..6477
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6474..6490
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6474..6490
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        6761..7140
                     /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 gccaccgctc ctggccctag cagcttcaga
      841 tgcctgcggc ctttcagcgc caccggcacc ttcaccaacg tggccaaggc ccaccccacc
      901 cagaactgcg actacttcta cttcagcgtg gaagatgcca aggccgagct gaagtgcgcc
      961 agcccttctg ccgccaccac cgccgtgtcc gacgagatga gctgcaacgt gaccgtgcgg
     1021 atgagcatgc ggagagccag cgacgtggtg ccccggctgc agagacaggt cggaaccgac
     1081 aaggactact tcacccacga ggccagcacc gacagcacca ccgccagccc caccagagtg
     1141 ggcaatacca gagcctccgc cgtgcccttc ctgaccgagg acgaggatgt gtggcgctac
     1201 gccgtgtccc tgaaggccaa gggcgacagc agcatccact acaagggctt caacggcggc
     1261 aacggctaca gcctgtgctg tgccgccctg gaagaggccg actgcgcctg gatgaccccc
     1321 cagaagggcg acaaggacct ggactgcgac gaggacgtgg acgtccgctt tgcccctcag
     1381 cggggcagag tgctgagagg ctgccccctg cctttcccag cccctagatc tgccggcgac
     1441 ggcgtgttcg gagcctctgc tggcgccaga gtgctgctga ccgacctgga tgagggcgag
     1501 ggctctggcg tgttccacgc cgtgatcacc aagcccctgc acagattcgt ggaaggcccc
     1561 tgggaagtga tggtgcagat gtggcggcgg agacagcaca tgcctagagt gggccgcgag
     1621 gacagcctga gcgtgcccgc cgatcacagc atcgaggccg aggtgctggg ccggatcatg
     1681 gtgcccttca ccctgaacct gaccgagctg cagaagcagg gcagaatcac ccagatgccc
     1741 agcatggccc tgacagtgga agatgtggcc gacgtggccc aggccgctgg cgagaaagct
     1801 gccgatgaag ccggcgacct gggcgcgccc ggcagcctgc accacatcct ggacgcccag
     1861 aagatgctgt ggaaccacag ggacaggaac ctgcctcctc tggcccctct gggcccccac
     1921 caccatcacc accattgagt ctagacacgt gattaattaa ggatcccccg acctcgacct
     1981 ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct
     2041 cactcggaag gacatatggg agggcaaatc atttggtcga gatccctcgg agatctctag
     2101 ctagaggatc gatccccgcc ccggacgaac taaacctgac tacgacatct ctgccccttc
     2161 ttcgcggggc agtgcatgta atcccttcag ttggttggta caacttgcca actgggccct
     2221 gttccacatg tgacacgggg ggggaccaaa cacaaagggg ttctctgact gtagttgaca
     2281 tccttataaa tggatgtgca catttgccaa cactgagtgg ctttcatcct ggagcagact
     2341 ttgcagtctg tggactgcaa cacaacattg cctttatgtg taactcttgg ctgaagctct
     2401 tacaccaatg ctgggggaca tgtacctccc aggggcccag gaagactacg ggaggctaca
     2461 ccaacgtcaa tcagaggggc ctgtgtagct accgataagc ggaccctcaa gagggcatta
     2521 gcaatagtgt ttataaggcc cccttgttaa ccctaaacgg gtagcatatg cttcccgggt
     2581 agtagtatat actatccaga ctaaccctaa ttcaatagca tatgttaccc aacgggaagc
     2641 atatgctatc gaattagggt tagtaaaagg gtcctaagga acagcgatat ctcccacccc
     2701 atgagctgtc acggttttat ttacatgggg tcaggattcc acgagggtag tgaaccattt
     2761 tagtcacaag ggcagtggct gaagatcaag gagcgggcag tgaactctcc tgaatcttcg
     2821 cctgcttctt cattctcctt cgtttagcta atagaataac tgctgagttg tgaacagtaa
     2881 ggtgtatgtg aggtgctcga aaacaaggtt tcaggtgacg cccccagaat aaaatttgga
     2941 cggggggttc agtggtggca ttgtgctatg acaccaatat aaccctcaca aaccccttgg
     3001 gcaataaata ctagtgtagg aatgaaacat tctgaatatc tttaacaata gaaatccatg
     3061 gggtggggac aagccgtaaa gactggatgt ccatctcaca cgaatttatg gctatgggca
     3121 acacataatc ctagtgcaat atgatactgg ggttattaag atgtgtccca ggcagggacc
     3181 aagacaggtg aaccatgttg ttacactcta tttgtaacaa ggggaaagag agtggacgcc
     3241 gacagcagcg gactccactg gttgtctcta acacccccga aaattaaacg gggctccacg
     3301 ccaatggggc ccataaacaa agacaagtgg ccactctttt ttttgaaatt gtggagtggg
     3361 ggcacgcgtc agcccccaca cgccgccctg cggttttgga ctgtaaaata agggtgtaat
     3421 aacttggctg attgtaaccc cgctaaccac tgcggtcaaa ccacttgccc acaaaaccac
     3481 taatggcacc ccggggaata cctgcataag taggtgggcg ggccaagata ggggcgcgat
     3541 tgctgcgatc tggaggacaa attacacaca cttgcgcctg agcgccaagc acagggttgt
     3601 tggtcctcat attcacgagg tcgctgagag cacggtgggc taatgttgcc atgggtagca
     3661 tatactaccc aaatatctgg atagcatatg ctatcctaat ctatatctgg gtagcatagg
     3721 ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagtatatg
     3781 ctatcctaat ttatatctgg gtagcatagg ctatcctaat ctatatctgg gtagcatatg
     3841 ctatcctaat ctatatctgg gtagtatatg ctatcctaat ctgtatccgg gtagcatatg
     3901 ctatcctaat agagattagg gtagtatatg ctatcctaat ttatatctgg gtagcatata
     3961 ctacccaaat atctggatag catatgctat cctaatctat atctgggtag catatgctat
     4021 cctaatctat atctgggtag cataggctat cctaatctat atctgggtag catatgctat
     4081 cctaatctat atctgggtag tatatgctat cctaatttat atctgggtag cataggctat
     4141 cctaatctat atctgggtag catatgctat cctaatctat atctgggtag tatatgctat
     4201 cctaatctgt atccgggtag catatgctat cctcatgata agctgtcaaa catgagaatt
     4261 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     4321 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     4381 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4441 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     4501 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     4561 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     4621 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     4681 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
     4741 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     4801 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     4861 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     4921 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     4981 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact
     5041 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     5101 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     5161 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     5221 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     5281 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     5341 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     5401 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     5461 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     5521 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     5581 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     5641 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     5701 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     5761 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     5821 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     5881 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     5941 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     6001 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     6061 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     6121 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     6181 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     6241 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     6301 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     6361 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     6421 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     6481 cagctatgac catgattacg ccaagctcta gctagaggtc gaccaattct catgtttgac
     6541 agcttatcat cgcagatccg ggcaacgttg ttgccattgc tgcaggcgca gaactggtag
     6601 gtatggaaga tctatacatt gaatcaatat tggcaattag ccatattagt cattggttat
     6661 atagcataaa tcaatattgg ctattggcca ttgcatacgt tgtatctata tcataatatg
     6721 tacatttata ttggctcatg tccaatatga ccgccatgtt gacattgatt attgactagt
     6781 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     6841 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     6901 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     6961 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     7021 ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     7081 accttacggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
//
