LOCUS       sequence-274736-        7320 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 7320)
  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 7320)
  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: D81_Ld_010280
FEATURES             Location/Qualifiers
     source          1..7320
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     1086..1105
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1128..1137
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2478..2495
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2547..2602
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2601..2620)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4838..4856)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4924..5028
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5029..5097,5098..5889)
                     /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(5247..5266)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6060..6648
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6549..6568
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6802..6819
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6936..6957
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6972..6977,6978..6995,6996..7002)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7010..7026
                     /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     7015..7037
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7034..7050
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7034..7050
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agatcctcac tctcttccgc atcgctgtct gcgagggcca gctgttgggc tcgcggttga
      661 ggacaaactc ttcgcggtct ttccagtact cttggatcgg aaacccgtcg gcctccgaac
      721 ggtactccgc caccgaggga cctgagcgag tccgcatcga ccggatcgga aaacctctcg
      781 agaaaggcgt ctaaccagtc acagtcgcaa ggtaggctga gcaccgtggc gggcggcagc
      841 gggtggcggt cggggttgtt tctggcggag gtgctgctga tgatgtaatt aaagtaggcg
      901 gtcttgagac ggcggatggt cgaggtgagg tgtggcaggc ttgagatcca gctgttgggg
      961 tgagtactcc ctctcaaaag cgggcattac ttctgcgcta agattgtcag tttccaaaaa
     1021 cgaggaggat ttgatattca cctggcccga tctggccata cacttgagtg acaatgacat
     1081 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caagtttgcc accatggagt
     1141 ttcagaccca ggtactcatg tccctgctgc tctgcatgtc tggtgcggcc gccctgtcag
     1201 aactggaacc cgccccgcag agagcccccg tggaaaagcg acgcagacag cacattccac
     1261 tgagtctcca ggcaagagaa gagctctcta ggaagagcaa tgagttgcag taccatgtgg
     1321 tcacccagga ctggtttggt cagagggtcg acgattttgt catgcaacat catcccgagt
     1381 gggattatga gacagtcaag aggctcgtac agcagggtca catatatcgg taccgaaaga
     1441 atggtaagaa gagatatact cggctcaccg accgtctcga gttcgacgag ctggtggtag
     1501 tccccaccgc atcattttgg gagaggcagc tggctccacc ctcaggggtc gaactgcagt
     1561 cctcaagcac tagtcagcag caggagcgca gacaaaagtt ccatctttct gcgaaaacaa
     1621 gggagatggc acaggaaatg gtactcttca agaacgaaca tgtcatcgtc atcaacaagc
     1681 ccagcggcgt gccaatcatg ccaacccacg acccactggc gatgaatatc acagacttgc
     1741 tgcctgcttg gcgatataca aacactcaga aacccgtgat ctgccataac ctggatactg
     1801 aaacttctgg gtgtgtcgtc ctggccagat ccgccaatgc ccatcggatg ctggggcgta
     1861 tgtttgttaa gagagtggta ccaaattccg tatactgggg ctttgcagtg ggaaagcccc
     1921 cagttaactt tggaagaatc cgaatgcact ttgagatcca gaaaggacaa ggtggcgatg
     1981 tgatagtggc ccggccgacc cccacagccg attctaaagt aggcatcgcc gagtttgttg
     2041 taaatgccag cgctctggag ttcggctcct ttatcagctt ttacccactg accactaggc
     2101 gccatcagga gcgaatcatg gcagcccacg ccctgagggc ccctttgctg ggtgacgcca
     2161 agtatggcgg ggaaagcgcc tttccacaca gcctgagctt gttctgggat cccgctcgaa
     2221 aggacgttcc gttgcatctg caccatcgca agatccagtt gccttataag aacggcgctg
     2281 gtgagttcgt gtgcgtgact gcacctctcc cgccccatat ggaaaagaca tttaaacgtc
     2341 tcgggtggcc tgtggacgcg gacgaccccc tcatccctgg cggcgcgccc ggcagcctgc
     2401 accacatcct ggacgcccag aagatgctgt ggaaccacag ggacaggaac ctgcctcctc
     2461 tggcccctct gggcccccac caccatcacc accattgagt ctagacacgt gattaattaa
     2521 ggatcccccg acctcgacct ctggctaata aaggaaattt attttcattg caatagtgtg
     2581 ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc atttggtcga
     2641 gatccctcgg agatctctag ctagaggatc gatccccgcc ccggacgaac taaacctgac
     2701 tacgacatct ctgccccttc ttcgcggggc agtgcatgta atcccttcag ttggttggta
     2761 caacttgcca actgggccct gttccacatg tgacacgggg ggggaccaaa cacaaagggg
     2821 ttctctgact gtagttgaca tccttataaa tggatgtgca catttgccaa cactgagtgg
     2881 ctttcatcct ggagcagact ttgcagtctg tggactgcaa cacaacattg cctttatgtg
     2941 taactcttgg ctgaagctct tacaccaatg ctgggggaca tgtacctccc aggggcccag
     3001 gaagactacg ggaggctaca ccaacgtcaa tcagaggggc ctgtgtagct accgataagc
     3061 ggaccctcaa gagggcatta gcaatagtgt ttataaggcc cccttgttaa ccctaaacgg
     3121 gtagcatatg cttcccgggt agtagtatat actatccaga ctaaccctaa ttcaatagca
     3181 tatgttaccc aacgggaagc atatgctatc gaattagggt tagtaaaagg gtcctaagga
     3241 acagcgatat ctcccacccc atgagctgtc acggttttat ttacatgggg tcaggattcc
     3301 acgagggtag tgaaccattt tagtcacaag ggcagtggct gaagatcaag gagcgggcag
     3361 tgaactctcc tgaatcttcg cctgcttctt cattctcctt cgtttagcta atagaataac
     3421 tgctgagttg tgaacagtaa ggtgtatgtg aggtgctcga aaacaaggtt tcaggtgacg
     3481 cccccagaat aaaatttgga cggggggttc agtggtggca ttgtgctatg acaccaatat
     3541 aaccctcaca aaccccttgg gcaataaata ctagtgtagg aatgaaacat tctgaatatc
     3601 tttaacaata gaaatccatg gggtggggac aagccgtaaa gactggatgt ccatctcaca
     3661 cgaatttatg gctatgggca acacataatc ctagtgcaat atgatactgg ggttattaag
     3721 atgtgtccca ggcagggacc aagacaggtg aaccatgttg ttacactcta tttgtaacaa
     3781 ggggaaagag agtggacgcc gacagcagcg gactccactg gttgtctcta acacccccga
     3841 aaattaaacg gggctccacg ccaatggggc ccataaacaa agacaagtgg ccactctttt
     3901 ttttgaaatt gtggagtggg ggcacgcgtc agcccccaca cgccgccctg cggttttgga
     3961 ctgtaaaata agggtgtaat aacttggctg attgtaaccc cgctaaccac tgcggtcaaa
     4021 ccacttgccc acaaaaccac taatggcacc ccggggaata cctgcataag taggtgggcg
     4081 ggccaagata ggggcgcgat tgctgcgatc tggaggacaa attacacaca cttgcgcctg
     4141 agcgccaagc acagggttgt tggtcctcat attcacgagg tcgctgagag cacggtgggc
     4201 taatgttgcc atgggtagca tatactaccc aaatatctgg atagcatatg ctatcctaat
     4261 ctatatctgg gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat
     4321 ctatatctgg gtagtatatg ctatcctaat ttatatctgg gtagcatagg ctatcctaat
     4381 ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat
     4441 ctgtatccgg gtagcatatg ctatcctaat agagattagg gtagtatatg ctatcctaat
     4501 ttatatctgg gtagcatata ctacccaaat atctggatag catatgctat cctaatctat
     4561 atctgggtag catatgctat cctaatctat atctgggtag cataggctat cctaatctat
     4621 atctgggtag catatgctat cctaatctat atctgggtag tatatgctat cctaatttat
     4681 atctgggtag cataggctat cctaatctat atctgggtag catatgctat cctaatctat
     4741 atctgggtag tatatgctat cctaatctgt atccgggtag catatgctat cctcatgata
     4801 agctgtcaaa catgagaatt ttcttgaaga cgaaagggcc tcgtgatacg cctattttta
     4861 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     4921 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     4981 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     5041 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     5101 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     5161 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     5221 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc
     5281 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     5341 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     5401 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     5461 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     5521 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg
     5581 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     5641 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     5701 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     5761 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     5821 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     5881 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     5941 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     6001 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     6061 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     6121 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     6181 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     6241 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     6301 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     6361 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     6421 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     6481 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     6541 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     6601 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     6661 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     6721 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     6781 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     6841 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     6901 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     6961 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     7021 taacaatttc acacaggaaa cagctatgac catgattacg ccaagctcta gctagaggtc
     7081 gaccaattct catgtttgac agcttatcat cgcagatccg ggcaacgttg ttgccattgc
     7141 tgcaggcgca gaactggtag gtatggaaga tctatacatt gaatcaatat tggcaattag
     7201 ccatattagt cattggttat atagcataaa tcaatattgg ctattggcca ttgcatacgt
     7261 tgtatctata tcataatatg tacatttata ttggctcatg tccaatatga ccgccatgtt
//
