LOCUS       sequence-274738-        9060 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 9060)
  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 9060)
  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: D80_Ld_220290
FEATURES             Location/Qualifiers
     source          1..9060
                     /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             3838..3855
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3907..3962
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3961..3980)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6198..6216)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6284..6388
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6389..6457,6458..7249)
                     /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(6607..6626)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7420..8008
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7909..7928
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8162..8179
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8296..8317
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8332..8337,8338..8355,8356..8362)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8370..8386
                     /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     8375..8397
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8394..8410
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8394..8410
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        8681..9060
                     /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 gcccggcctt gtacgaacgc cgacattatc
      841 cacttcttct ctccttgtga caagcgaacg ggaaagctga ccgtgttgag ctaccttagc
      901 tcagatgcgg attgcgagcg cgggaacctg tttgttcctc ctcccaggac aacagcatgc
      961 tatgactgca ccgatggata tcggctggat cctctcacac ttgaatgcgc cgtgtgtcca
     1021 gccggtactt attctagtta tacgactagg tacgtgacct tcgatccctt gcctgaggac
     1081 atgcgcactt actgttcccc agaaccatgt gcatcatggt ccgccgtgga agacacctat
     1141 ggcactgttg tcagcagcgg ggtccagccc actgagaatc gccctgttac ccaatggggc
     1201 accatcgacg aaagtgttga gtccacgctg gatttcgttg tcgatgttat agaatccagc
     1261 ggctccctgg agttcgatta ctccatctca agtgaggaga tgtacgacgg actggtgctc
     1321 cgactcaatt acagcatcgt gcgaaatcct gataccgagg tgggaaaata tcggttcttc
     1381 gcaaccggag tgcatagaga ctggtaccac gctagcctga gactccccta tgggaccact
     1441 caaatacaat tcgattacat gaaggatagt actgggcccc cgtatggcga agagctcact
     1501 ggtgtgaatg acagagcact gttgcgcaat atcgtagtta aaggtacaag gaagaccgtc
     1561 gcagaaatgc agtgcgtttc ttgtccaccc ggccactgga cggcaagaga ggggagcggt
     1621 ggtgtcgggg ccagtcaatg cgtcaggtgt ccaagaaaca catacagggg cgaaggggac
     1681 acccagtgtc tgcagtgtcc acccaacaca tcaagcccag aaggggctaa taattgtact
     1741 cgcatgagag cttgtaccgg cggcgattac atcgcacaat acggttcatg ccgtagcgac
     1801 aataccagat tgagatcatg ggtgaaatac gaaggcgctg attgtgcgga agtggaaggt
     1861 tcacgcccta cagctacccg ggtggaatgc gctacctgtc tgcccggaat gcggcatgct
     1921 gatagtaggg ctcgcggcgc ttgcgtttca tgtcctgccg gggaatattt gctcggagac
     1981 aggtgtgagg tgtgcccctt ggggacagcc gctataccca agcttgtgta cagttccggg
     2041 ttcgatggtc tgggcggcga cgtgatggcc ttgccgggga tctctctgag cagaatttac
     2101 tgcactgggt ccgcctctcg ctgtcgccgg gaagagggta aggggttcga actcgtgggc
     2161 cttccatttg cagatgagga caacactctg ataggcatcc gccaggagca cctcgcctct
     2221 tccacatact ctctggctac tctgagctac acctttcacg ccgaagagga cggctgggcg
     2281 aatattacct ttgcttatgt cgacgctgaa ggaaagtatg tccaacgcat ggactggcgc
     2341 accactctga atgtgcgagt aattgtggat gataacgcag agtatcccgt gagggctttt
     2401 gcgtctcaca ctggccgtgt ggattccgtg ttctacagcc tgcttgtccc ttattatgtg
     2461 gataccgcac tgcctgcttc cggatctgat agcaaattga gaggcatgcg tcatcggctg
     2521 tcttggatcg tggaagaatt ggccttcaca tctaatccaa tcagtgttgt gatagtcgga
     2581 ctggaagtct tcggcgacgt ctcaggtggg gtgcagacct gccagccatg cagggccggg
     2641 ttcagttgtg ctagtagggc ggctaagatg acaccttgct cccccggtac tgtgcagccg
     2701 gcactcaggg ctcatcgctg cgagccttgt atagggaaca catacgcccc tggctatggg
     2761 tttactacct gcctgcagtg cccggggaac actgtcgcca atgcagacca taccggatgc
     2821 tccttgggct gtgtatttga gcgcaacggc atggtctata acttcagtga gcttcgcggc
     2881 gtggtactca acgccagtgt cggctacatg cctgccacta acgcttcagt cagcaacatg
     2941 actgtggaag aggccgatga agctggtata ccacggtttt acttttctcc ttgcgatcca
     3001 ctgcccgtta gagacgcgcg cgcgacgagc gaatccgact ctgcgcttac ccctatcacg
     3061 agagatagac tgtgcgtccc gtctgccaga ggcaggaaca gctccagctc tgcctacgtt
     3121 tgtcagcgca cctctgcaac aaatgggcat cattttggag acgccgtggt tcatatggaa
     3181 gtaggaagac acctctatct ggtgacgcaa ttgggctccc cgtatatttt ctacggtgtt
     3241 gatacatccg cagacgattg ggtgtatgac gctaagtcca attacatggc attcattgag
     3301 ctggaatgta gcacatctgg tggcgagcct aaatttggta cactgcagta cgtgggtgaa
     3361 gggaatgagg acctggtgct caggtggcgt tccgcctatg cctgccccct ttgtacaaat
     3421 gaatcatacg agaaggtgga aacttcatgc gatgatccta cggccacaac cagaagggtc
     3481 tactatacac tgcgccgcga ctctatgaat tgtgtgggcg ggtacgtccc acccgaaccg
     3541 ataattgtta agtgtactga gtgcaccctg gatgcttaca ctgttagatg gctggaatgt
     3601 gatccctcaa ctcaaactca ggaaggggtg tatgtgttgt tgccggatcg ggcgaactgc
     3661 acccctggag ctagtcctct tccacccgta aagccaagac cgtgtgatgc ttccggtggt
     3721 actgccggac gtggcaccag cggcgcgccc ggcagcctgc accacatcct ggacgcccag
     3781 aagatgctgt ggaaccacag ggacaggaac ctgcctcctc tggcccctct gggcccccac
     3841 caccatcacc accattgagt ctagacacgt gattaattaa ggatcccccg acctcgacct
     3901 ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct
     3961 cactcggaag gacatatggg agggcaaatc atttggtcga gatccctcgg agatctctag
     4021 ctagaggatc gatccccgcc ccggacgaac taaacctgac tacgacatct ctgccccttc
     4081 ttcgcggggc agtgcatgta atcccttcag ttggttggta caacttgcca actgggccct
     4141 gttccacatg tgacacgggg ggggaccaaa cacaaagggg ttctctgact gtagttgaca
     4201 tccttataaa tggatgtgca catttgccaa cactgagtgg ctttcatcct ggagcagact
     4261 ttgcagtctg tggactgcaa cacaacattg cctttatgtg taactcttgg ctgaagctct
     4321 tacaccaatg ctgggggaca tgtacctccc aggggcccag gaagactacg ggaggctaca
     4381 ccaacgtcaa tcagaggggc ctgtgtagct accgataagc ggaccctcaa gagggcatta
     4441 gcaatagtgt ttataaggcc cccttgttaa ccctaaacgg gtagcatatg cttcccgggt
     4501 agtagtatat actatccaga ctaaccctaa ttcaatagca tatgttaccc aacgggaagc
     4561 atatgctatc gaattagggt tagtaaaagg gtcctaagga acagcgatat ctcccacccc
     4621 atgagctgtc acggttttat ttacatgggg tcaggattcc acgagggtag tgaaccattt
     4681 tagtcacaag ggcagtggct gaagatcaag gagcgggcag tgaactctcc tgaatcttcg
     4741 cctgcttctt cattctcctt cgtttagcta atagaataac tgctgagttg tgaacagtaa
     4801 ggtgtatgtg aggtgctcga aaacaaggtt tcaggtgacg cccccagaat aaaatttgga
     4861 cggggggttc agtggtggca ttgtgctatg acaccaatat aaccctcaca aaccccttgg
     4921 gcaataaata ctagtgtagg aatgaaacat tctgaatatc tttaacaata gaaatccatg
     4981 gggtggggac aagccgtaaa gactggatgt ccatctcaca cgaatttatg gctatgggca
     5041 acacataatc ctagtgcaat atgatactgg ggttattaag atgtgtccca ggcagggacc
     5101 aagacaggtg aaccatgttg ttacactcta tttgtaacaa ggggaaagag agtggacgcc
     5161 gacagcagcg gactccactg gttgtctcta acacccccga aaattaaacg gggctccacg
     5221 ccaatggggc ccataaacaa agacaagtgg ccactctttt ttttgaaatt gtggagtggg
     5281 ggcacgcgtc agcccccaca cgccgccctg cggttttgga ctgtaaaata agggtgtaat
     5341 aacttggctg attgtaaccc cgctaaccac tgcggtcaaa ccacttgccc acaaaaccac
     5401 taatggcacc ccggggaata cctgcataag taggtgggcg ggccaagata ggggcgcgat
     5461 tgctgcgatc tggaggacaa attacacaca cttgcgcctg agcgccaagc acagggttgt
     5521 tggtcctcat attcacgagg tcgctgagag cacggtgggc taatgttgcc atgggtagca
     5581 tatactaccc aaatatctgg atagcatatg ctatcctaat ctatatctgg gtagcatagg
     5641 ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagtatatg
     5701 ctatcctaat ttatatctgg gtagcatagg ctatcctaat ctatatctgg gtagcatatg
     5761 ctatcctaat ctatatctgg gtagtatatg ctatcctaat ctgtatccgg gtagcatatg
     5821 ctatcctaat agagattagg gtagtatatg ctatcctaat ttatatctgg gtagcatata
     5881 ctacccaaat atctggatag catatgctat cctaatctat atctgggtag catatgctat
     5941 cctaatctat atctgggtag cataggctat cctaatctat atctgggtag catatgctat
     6001 cctaatctat atctgggtag tatatgctat cctaatttat atctgggtag cataggctat
     6061 cctaatctat atctgggtag catatgctat cctaatctat atctgggtag tatatgctat
     6121 cctaatctgt atccgggtag catatgctat cctcatgata agctgtcaaa catgagaatt
     6181 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     6241 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     6301 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     6361 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     6421 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     6481 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     6541 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     6601 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
     6661 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     6721 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     6781 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     6841 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     6901 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact
     6961 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     7021 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     7081 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     7141 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     7201 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     7261 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     7321 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     7381 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     7441 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     7501 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     7561 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     7621 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     7681 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     7741 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     7801 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     7861 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     7921 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     7981 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     8041 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     8101 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     8161 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     8221 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     8281 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     8341 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     8401 cagctatgac catgattacg ccaagctcta gctagaggtc gaccaattct catgtttgac
     8461 agcttatcat cgcagatccg ggcaacgttg ttgccattgc tgcaggcgca gaactggtag
     8521 gtatggaaga tctatacatt gaatcaatat tggcaattag ccatattagt cattggttat
     8581 atagcataaa tcaatattgg ctattggcca ttgcatacgt tgtatctata tcataatatg
     8641 tacatttata ttggctcatg tccaatatga ccgccatgtt gacattgatt attgactagt
     8701 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     8761 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     8821 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     8881 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     8941 ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     9001 accttacggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
//
