LOCUS       sequence-262329-        7734 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 7734)
  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 7734)
  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: D36_Ld_323600
FEATURES             Location/Qualifiers
     source          1..7734
                     /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             2898..2915
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2967..3022
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3021..3040)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5258..5276)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5344..5448
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5449..5517,5518..6309)
                     /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(5667..5686)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6480..7068
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6969..6988
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7222..7239
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7356..7377
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7392..7397,7398..7415,7416..7422)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7430..7446
                     /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     7435..7457
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7454..7470
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7454..7470
                     /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 cctctggatc tgccaccgtg ggagcccaca acgacagact gccccagccc agctggtcca
     1261 gaacctgcac cagcccctac gaagtgtgca gcaccatcga ggtgtaccag tactgggagg
     1321 aaaagaagaa ccggctgagc ttcaccgtgg acgccatgat gaaggacccc aacgccaaga
     1381 acgtgctgac cgtgtggatg gtggtggacg acgccgacgt gaacatggcc aagggcatct
     1441 tccgcgaggg catcaacgtg gtcatcgtgc accccagagg cagcaagcag acccagagct
     1501 tcatgacctg cgccaacacc agcaacccct acaaccggca gtgctggatg agcgaggcct
     1561 gtgccgccga actggccgct gccgccagca gcacctccaa ccccctgaac atgtcccact
     1621 acagcgccaa gcaggccgcc cacgacctgg actgggctct gagaacactg ggcgacggca
     1681 gacctaacgt ggtgctgggc cagggcctga gcagcatgct ggtgctgcgg ctgctgcagt
     1741 accaccagga caccaaggcc gccgtgatcc tgctggacta tgtgcaccct gtgatgttcg
     1801 acgtgtacaa ctacttcgga ggcggcagca tggtcaccgc cctgcagcat ctgctggcca
     1861 tgtgcgacga ccaggccgct tgcgtgggca gactgggagc cgccgagggc agctggaaca
     1921 gactgatgac agtgatccat atggccaaag agggcaagct gagctgcgcc tccagactga
     1981 gatgggcccc cagaagaggc ggaaacgccg ccttcgccga cgagctgaga ggcgtgctgg
     2041 ccctgatgct gagatacccc gtgtacccct tcctggcctc caacgccgct ctgctgaacc
     2101 tgatccccag cctgctgtac cgggtgcagc ggtgcaacaa gaaagacgtg gccgcactga
     2161 acaagctgta cgactacctg agcgccgcca aaaactacga gtgccccgag agcgtggccc
     2221 tgcacatgca ctggctggtc aacgagttcc tgcaggctgc cccccctgcc aacatccaca
     2281 gcttcagaag cagcgccgca gccaagcacc tgattctgcc cagcttccac gccatgtccg
     2341 ccttccacag cgctgccaag aagttccccc ggatcagcag atccgccgag tccaagcagc
     2401 tgcccgccaa tgccacccag cctattctgc tgctgaatgc cgacaccgac gccctgctgc
     2461 ctggcggagc tgcttctcag gtggcactgg ccttcagaca gctgggcgat agcgtgcagc
     2521 tgaggcagct gcggggcatc atgaacaggc ccagcgctgt gctgacaccc tgcctcgtga
     2581 acaacctgaa gatgttccgg gaccacggcg tgtgggccga tgaggaccag tgtaccctgg
     2641 atgtggccca caagatcgac ttcatcaacg gcgccaccga catctactac ggcaccagcg
     2701 acgcctggga cttcgacaag cccaacagcg aggaaacaaa cggcggcagg gacgacaacg
     2761 acgacgaggg cggcttccac ttcaagtggc ggtttatcgc tggcgcgccc ggcagcctgc
     2821 accacatcct ggacgcccag aagatgctgt ggaaccacag ggacaggaac ctgcctcctc
     2881 tggcccctct gggcccccac caccatcacc accattgagt ctagacacgt gattaattaa
     2941 ggatcccccg acctcgacct ctggctaata aaggaaattt attttcattg caatagtgtg
     3001 ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc atttggtcga
     3061 gatccctcgg agatctctag ctagaggatc gatccccgcc ccggacgaac taaacctgac
     3121 tacgacatct ctgccccttc ttcgcggggc agtgcatgta atcccttcag ttggttggta
     3181 caacttgcca actgggccct gttccacatg tgacacgggg ggggaccaaa cacaaagggg
     3241 ttctctgact gtagttgaca tccttataaa tggatgtgca catttgccaa cactgagtgg
     3301 ctttcatcct ggagcagact ttgcagtctg tggactgcaa cacaacattg cctttatgtg
     3361 taactcttgg ctgaagctct tacaccaatg ctgggggaca tgtacctccc aggggcccag
     3421 gaagactacg ggaggctaca ccaacgtcaa tcagaggggc ctgtgtagct accgataagc
     3481 ggaccctcaa gagggcatta gcaatagtgt ttataaggcc cccttgttaa ccctaaacgg
     3541 gtagcatatg cttcccgggt agtagtatat actatccaga ctaaccctaa ttcaatagca
     3601 tatgttaccc aacgggaagc atatgctatc gaattagggt tagtaaaagg gtcctaagga
     3661 acagcgatat ctcccacccc atgagctgtc acggttttat ttacatgggg tcaggattcc
     3721 acgagggtag tgaaccattt tagtcacaag ggcagtggct gaagatcaag gagcgggcag
     3781 tgaactctcc tgaatcttcg cctgcttctt cattctcctt cgtttagcta atagaataac
     3841 tgctgagttg tgaacagtaa ggtgtatgtg aggtgctcga aaacaaggtt tcaggtgacg
     3901 cccccagaat aaaatttgga cggggggttc agtggtggca ttgtgctatg acaccaatat
     3961 aaccctcaca aaccccttgg gcaataaata ctagtgtagg aatgaaacat tctgaatatc
     4021 tttaacaata gaaatccatg gggtggggac aagccgtaaa gactggatgt ccatctcaca
     4081 cgaatttatg gctatgggca acacataatc ctagtgcaat atgatactgg ggttattaag
     4141 atgtgtccca ggcagggacc aagacaggtg aaccatgttg ttacactcta tttgtaacaa
     4201 ggggaaagag agtggacgcc gacagcagcg gactccactg gttgtctcta acacccccga
     4261 aaattaaacg gggctccacg ccaatggggc ccataaacaa agacaagtgg ccactctttt
     4321 ttttgaaatt gtggagtggg ggcacgcgtc agcccccaca cgccgccctg cggttttgga
     4381 ctgtaaaata agggtgtaat aacttggctg attgtaaccc cgctaaccac tgcggtcaaa
     4441 ccacttgccc acaaaaccac taatggcacc ccggggaata cctgcataag taggtgggcg
     4501 ggccaagata ggggcgcgat tgctgcgatc tggaggacaa attacacaca cttgcgcctg
     4561 agcgccaagc acagggttgt tggtcctcat attcacgagg tcgctgagag cacggtgggc
     4621 taatgttgcc atgggtagca tatactaccc aaatatctgg atagcatatg ctatcctaat
     4681 ctatatctgg gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat
     4741 ctatatctgg gtagtatatg ctatcctaat ttatatctgg gtagcatagg ctatcctaat
     4801 ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat
     4861 ctgtatccgg gtagcatatg ctatcctaat agagattagg gtagtatatg ctatcctaat
     4921 ttatatctgg gtagcatata ctacccaaat atctggatag catatgctat cctaatctat
     4981 atctgggtag catatgctat cctaatctat atctgggtag cataggctat cctaatctat
     5041 atctgggtag catatgctat cctaatctat atctgggtag tatatgctat cctaatttat
     5101 atctgggtag cataggctat cctaatctat atctgggtag catatgctat cctaatctat
     5161 atctgggtag tatatgctat cctaatctgt atccgggtag catatgctat cctcatgata
     5221 agctgtcaaa catgagaatt ttcttgaaga cgaaagggcc tcgtgatacg cctattttta
     5281 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     5341 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     5401 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     5461 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     5521 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     5581 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     5641 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc
     5701 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     5761 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     5821 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     5881 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     5941 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg
     6001 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     6061 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     6121 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     6181 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     6241 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     6301 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     6361 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     6421 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     6481 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     6541 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     6601 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     6661 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     6721 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     6781 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     6841 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     6901 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     6961 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     7021 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     7081 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     7141 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     7201 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     7261 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     7321 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     7381 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     7441 taacaatttc acacaggaaa cagctatgac catgattacg ccaagctcta gctagaggtc
     7501 gaccaattct catgtttgac agcttatcat cgcagatccg ggcaacgttg ttgccattgc
     7561 tgcaggcgca gaactggtag gtatggaaga tctatacatt gaatcaatat tggcaattag
     7621 ccatattagt cattggttat atagcataaa tcaatattgg ctattggcca ttgcatacgt
     7681 tgtatctata tcataatatg tacatttata ttggctcatg tccaatatga ccgc
//
