LOCUS       sequence-262338-        8424 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 8424)
  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 8424)
  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: D45_Ld_343960
FEATURES             Location/Qualifiers
     source          1..8424
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        7..386
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        387..590
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     541..561
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     587..611
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     1092..1111
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1134..1143
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3588..3605
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3657..3712
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3711..3730)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5948..5966)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6034..6138
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6139..6207,6208..6999)
                     /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(6357..6376)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7170..7758
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7659..7678
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7912..7929
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8046..8067
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8082..8087,8088..8105,8106..8112)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8120..8136
                     /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     8125..8147
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8144..8160
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8144..8160
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 catgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
       61 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      121 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      181 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      241 tacatcaagt gtatcatatg ccaagtccgc cccctattga cgtcaatgac ggtaaatggc
      301 ccgcctggca ttatgcccag tacatgacct tacgggactt tcctacttgg cagtacatct
      361 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacacc aatgggcgtg
      421 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      481 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taataacccc gccccgttga
      541 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga
      601 accgtcagat cctcactctc ttccgcatcg ctgtctgcga gggccagctg ttgggctcgc
      661 ggttgaggac aaactcttcg cggtctttcc agtactcttg gatcggaaac ccgtcggcct
      721 ccgaacggta ctccgccacc gagggacctg agcgagtccg catcgaccgg atcggaaaac
      781 ctctcgagaa aggcgtctaa ccagtcacag tcgcaaggta ggctgagcac cgtggcgggc
      841 ggcagcgggt ggcggtcggg gttgtttctg gcggaggtgc tgctgatgat gtaattaaag
      901 taggcggtct tgagacggcg gatggtcgag gtgaggtgtg gcaggcttga gatccagctg
      961 ttggggtgag tactccctct caaaagcggg cattacttct gcgctaagat tgtcagtttc
     1021 caaaaacgag gaggatttga tattcacctg gcccgatctg gccatacact tgagtgacaa
     1081 tgacatccac tttgcctttc tctccacagg tgtccactcc caggtccaag tttgccacca
     1141 tggagtttca gacccaggta ctcatgtccc tgctgctctg catgtctggt gcggccgcca
     1201 ccatccacga ggatgagcag ggcctgcggg actggatcat gcacctcgtg ggcaacgtgc
     1261 aggactctgc cgcccaggtg gccaccaacc cccagctgct gtacgtggcc tctgaggatg
     1321 gcgccgtggc cgccattgcc gtgggagcct atggcggcct gaacctgacc tggcggcaga
     1381 tcagcagcgc cacccctctg tgtgtggccg ctggacctca ggccgtgctg accgtgaaca
     1441 acgccggcgt ggccacactg atgagcccca ccaccggcag catcgaagtg acctacgccc
     1501 tgcagaccgt gaccaccaca ctgcaggccg ctgcctgtag cgtggtgtcc ggaaaggccg
     1561 tggtggtcac ctacgacggg gccaccctga agagattcga gttcagcacc gccagcgagg
     1621 aacagagcat ccccgtggct gcctacgcca gcgcctctgg cgaggtgtcc aagatgtacg
     1681 tcgccggcag cgtgctgctg gtcaaccacg gcagcgacag cgccgacgtg ctgagactgg
     1741 acaacctggc caaagaacgc agcgtggccg gaatggccac ctctatcgcc gccgacggcc
     1801 acttcaccac ccgggatgcc gccacagtgc ggagcttccc taccaacggc gctgccagcg
     1861 aagtggcctg cagcgactgt ggaatcgccg tcgtgcggaa cgcccacacc ggcgaaccta
     1921 ccggcgtcgt cagagtggaa acacaggtgg acatgctgcg gatcacctac cccaacagcg
     1981 tcgtgaccat ccccaacatc tgcagcagca tcaccgccta ccccctgctg gccttcgaga
     2041 aggacgacgg cgacgtgatc gtgctgatca agacaggcgc ccacaacctg ctgctcgtgt
     2101 ccgccgctga aggcctcgtg tggcggagat ttgaggccct ggctaacgtg gcagccgccg
     2161 ctatcgtgga acccatggaa gccctggacc acttccactt caacaagaac atcctgctgg
     2221 tgtccagatt cggcaccctg tacagcatcc ctattgccgg catgggagcc caggtcgact
     2281 acatcaagga cttcagccag gaactggctg ccgccatgaa ggcccccagc atggctaagg
     2341 tgcaggtccg agagctggcc gtgcgggatg atggcagaac cgctgtggtg cacgccgagt
     2401 ctggcgccac aagcggccac atcctgatcg acctggccga gcggatggtc accagcgtga
     2461 caacatgcga gaacgctgtg ctgagcaccc ccgagctgga agtggctgct gacctgagcg
     2521 tgcggggcag cctgagccac ggcgtgttct acacctatgc cagccacgcc gaaagcggca
     2581 ccatcgaggg ctactccgtg tctgctgccg caggcgccag acccacctgg gtggtgcaga
     2641 tgcctagcgc cattgtggcc cacgccgctg gcagcgagcc tcggagaacc gacgtggtca
     2701 acaacctgag agtgtacccc aatatcagcg gcagggaaat ggtggaagaa gtgcggcgga
     2761 cataccccat gcggaatgtg atcgctgtgg ctcactacga gccctccgag gaagaactgc
     2821 ccagcctggt catcacagcc atcgatgtgg tcacaggctc catcctggcc acaaccagac
     2881 acgccaacgt cgagggggac gtgaagatgg tcattgtgga acacgccatc gtgtactact
     2941 acctggacgc caagaagatg cggtactgct tcggcgtgtg ggagctgttc gaggacgaga
     3001 acagccctgt ggtgtctaag acagccggcg ctaccatccc tcagattatc gccagcttct
     3061 tcgtgaacac caagagagag ttctccagcc gggccaccag gccccccatc gtggtcgtgt
     3121 ctaccctggg cagctttggc ggccctctgg ccgatatggg agtgaccaca agctacaacg
     3181 ctatcgcccg gaagtccctg gtgctggctt acgccacagg ccgggtcgtc gtggtggaac
     3241 tgagacacct cctggccggg aaccagatgc ccctgcctgg ggccaaggac cggcagatga
     3301 gccatctgct gctgcctagc ttcctgttcg cctcccacaa gtaccgggtg gccttcccca
     3361 gaaagatcac caccgagccc accctgctcg agagcagctg tcatgtggtc gtcagcggcc
     3421 tggatctgtt ctacgtgcgg tccagcagcg gcaagccctt cgacctgctg aacagcgact
     3481 ttaacaagcc tggcgcgccc ggcagcctgc accacatcct ggacgcccag aagatgctgt
     3541 ggaaccacag ggacaggaac ctgcctcctc tggcccctct gggcccccac caccatcacc
     3601 accattgagt ctagacacgt gattaattaa ggatcccccg acctcgacct ctggctaata
     3661 aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag
     3721 gacatatggg agggcaaatc atttggtcga gatccctcgg agatctctag ctagaggatc
     3781 gatccccgcc ccggacgaac taaacctgac tacgacatct ctgccccttc ttcgcggggc
     3841 agtgcatgta atcccttcag ttggttggta caacttgcca actgggccct gttccacatg
     3901 tgacacgggg ggggaccaaa cacaaagggg ttctctgact gtagttgaca tccttataaa
     3961 tggatgtgca catttgccaa cactgagtgg ctttcatcct ggagcagact ttgcagtctg
     4021 tggactgcaa cacaacattg cctttatgtg taactcttgg ctgaagctct tacaccaatg
     4081 ctgggggaca tgtacctccc aggggcccag gaagactacg ggaggctaca ccaacgtcaa
     4141 tcagaggggc ctgtgtagct accgataagc ggaccctcaa gagggcatta gcaatagtgt
     4201 ttataaggcc cccttgttaa ccctaaacgg gtagcatatg cttcccgggt agtagtatat
     4261 actatccaga ctaaccctaa ttcaatagca tatgttaccc aacgggaagc atatgctatc
     4321 gaattagggt tagtaaaagg gtcctaagga acagcgatat ctcccacccc atgagctgtc
     4381 acggttttat ttacatgggg tcaggattcc acgagggtag tgaaccattt tagtcacaag
     4441 ggcagtggct gaagatcaag gagcgggcag tgaactctcc tgaatcttcg cctgcttctt
     4501 cattctcctt cgtttagcta atagaataac tgctgagttg tgaacagtaa ggtgtatgtg
     4561 aggtgctcga aaacaaggtt tcaggtgacg cccccagaat aaaatttgga cggggggttc
     4621 agtggtggca ttgtgctatg acaccaatat aaccctcaca aaccccttgg gcaataaata
     4681 ctagtgtagg aatgaaacat tctgaatatc tttaacaata gaaatccatg gggtggggac
     4741 aagccgtaaa gactggatgt ccatctcaca cgaatttatg gctatgggca acacataatc
     4801 ctagtgcaat atgatactgg ggttattaag atgtgtccca ggcagggacc aagacaggtg
     4861 aaccatgttg ttacactcta tttgtaacaa ggggaaagag agtggacgcc gacagcagcg
     4921 gactccactg gttgtctcta acacccccga aaattaaacg gggctccacg ccaatggggc
     4981 ccataaacaa agacaagtgg ccactctttt ttttgaaatt gtggagtggg ggcacgcgtc
     5041 agcccccaca cgccgccctg cggttttgga ctgtaaaata agggtgtaat aacttggctg
     5101 attgtaaccc cgctaaccac tgcggtcaaa ccacttgccc acaaaaccac taatggcacc
     5161 ccggggaata cctgcataag taggtgggcg ggccaagata ggggcgcgat tgctgcgatc
     5221 tggaggacaa attacacaca cttgcgcctg agcgccaagc acagggttgt tggtcctcat
     5281 attcacgagg tcgctgagag cacggtgggc taatgttgcc atgggtagca tatactaccc
     5341 aaatatctgg atagcatatg ctatcctaat ctatatctgg gtagcatagg ctatcctaat
     5401 ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat
     5461 ttatatctgg gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat
     5521 ctatatctgg gtagtatatg ctatcctaat ctgtatccgg gtagcatatg ctatcctaat
     5581 agagattagg gtagtatatg ctatcctaat ttatatctgg gtagcatata ctacccaaat
     5641 atctggatag catatgctat cctaatctat atctgggtag catatgctat cctaatctat
     5701 atctgggtag cataggctat cctaatctat atctgggtag catatgctat cctaatctat
     5761 atctgggtag tatatgctat cctaatttat atctgggtag cataggctat cctaatctat
     5821 atctgggtag catatgctat cctaatctat atctgggtag tatatgctat cctaatctgt
     5881 atccgggtag catatgctat cctcatgata agctgtcaaa catgagaatt ttcttgaaga
     5941 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     6001 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     6061 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     6121 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     6181 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     6241 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     6301 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     6361 tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac
     6421 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     6481 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     6541 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     6601 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     6661 gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc
     6721 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     6781 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     6841 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     6901 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     6961 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     7021 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     7081 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     7141 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     7201 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     7261 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     7321 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     7381 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     7441 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     7501 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     7561 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     7621 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     7681 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     7741 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     7801 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     7861 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     7921 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     7981 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     8041 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     8101 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     8161 catgattacg ccaagctcta gctagaggtc gaccaattct catgtttgac agcttatcat
     8221 cgcagatccg ggcaacgttg ttgccattgc tgcaggcgca gaactggtag gtatggaaga
     8281 tctatacatt gaatcaatat tggcaattag ccatattagt cattggttat atagcataaa
     8341 tcaatattgg ctattggcca ttgcatacgt tgtatctata tcataatatg tacatttata
     8401 ttggctcatg tccaatatga ccgc
//
