LOCUS       sequence-274900-        7470 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 7470)
  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 7470)
  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: D37_Ld_361830
FEATURES             Location/Qualifiers
     source          1..7470
                     /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             2248..2265
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2317..2372
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2371..2390)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4608..4626)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4694..4798
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4799..4867,4868..5659)
                     /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(5017..5036)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5830..6418
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6319..6338
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6572..6589
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6706..6727
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6742..6747,6748..6765,6766..6772)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6780..6796
                     /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     6785..6807
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6804..6820
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6804..6820
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7091..7470
                     /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 gccaccctgt gccccagcaa tggcaacacc
      841 cacaccgtgt acgtgaccag caacatgggc gacaacatct ggtccttcga caccctgggc
      901 aactacctgg gccccgtgct gaacaaggcc agcttcccac ccggcgtgcg ggtggacaag
      961 ctgcggagca tgagattcgg gccccagggc cacctgtatg tgtccagcgc cagaggcagc
     1021 tacagccggg tgttcgccgt gtccggcaac ggcctgctga acggcaccat ggaagagaac
     1081 tgcacccggg actacctgtt caccgtgatc gagcaggaca aagagaaccc cttcctggac
     1141 cacccctacg acatcacctt ccaccccgcc accgaggata tgtacgtgtc caaccagaac
     1201 agcgtgaccg tgacccggta caccagatac cggaccgccg acgtgacccc caaagccggc
     1261 gctggcggaa gccagcacgc cttctatcct gcctggaagc ccgccaagaa cgtgaaggcc
     1321 gccgtggaag ccgacgagga aaccgtgtct gccgaagaga cagtggtggc ccccaagaat
     1381 atccccgaca gcagcggcct gttcgccagc agctggtcca gctcctacag catgtcctcc
     1441 gtgcggggcc tggccatcag ccctctgctg cctagagccc tggtggaagg cggagccccc
     1501 agcggagtgt ttgccgtgac caacgagagc accatgtact acctgctcgt gtgcgacgtg
     1561 tcctccaata ccgtgcacgt gttcgacccc gagagcggcg agagactgtt cggcctgaac
     1621 gtgcccagcc ccatccaggt catgttcccc aaccggtact acatcagcga cagctccagc
     1681 ctgagcgtgt ccgccgctgg cgtgcacacc cagagattcg acgtgcccta catctacgtg
     1741 acatccaaag aggacggcat ggcctacatg gtccgattca gcagccggca gggcaatacc
     1801 aacaacaacg gcatgcaggg gggagccttc ggcgaggact tcatccgggc ccaccggctg
     1861 tacaccatca accaccctac ccccctgcac gccgccagcg gcatctacga gaatagcgcc
     1921 agggacgtgc tgctgatcgc cgaccggaac ggcagacgga tctacagcta cgccagccct
     1981 ttcctgaccg actacaccag cggctacggc ccctccccct ttctgggctt cttcgtgaag
     2041 cacctccccg accagcccga gttcatcatg agcaccctgc tggaacacca ggaaaacatc
     2101 ccattctgct acgaactcaa caaggacggg aagttcagat acgtggccct gggcgcgccc
     2161 ggcagcctgc accacatcct ggacgcccag aagatgctgt ggaaccacag ggacaggaac
     2221 ctgcctcctc tggcccctct gggcccccac caccatcacc accattgagt ctagacacgt
     2281 gattaattaa ggatcccccg acctcgacct ctggctaata aaggaaattt attttcattg
     2341 caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc
     2401 atttggtcga gatccctcgg agatctctag ctagaggatc gatccccgcc ccggacgaac
     2461 taaacctgac tacgacatct ctgccccttc ttcgcggggc agtgcatgta atcccttcag
     2521 ttggttggta caacttgcca actgggccct gttccacatg tgacacgggg ggggaccaaa
     2581 cacaaagggg ttctctgact gtagttgaca tccttataaa tggatgtgca catttgccaa
     2641 cactgagtgg ctttcatcct ggagcagact ttgcagtctg tggactgcaa cacaacattg
     2701 cctttatgtg taactcttgg ctgaagctct tacaccaatg ctgggggaca tgtacctccc
     2761 aggggcccag gaagactacg ggaggctaca ccaacgtcaa tcagaggggc ctgtgtagct
     2821 accgataagc ggaccctcaa gagggcatta gcaatagtgt ttataaggcc cccttgttaa
     2881 ccctaaacgg gtagcatatg cttcccgggt agtagtatat actatccaga ctaaccctaa
     2941 ttcaatagca tatgttaccc aacgggaagc atatgctatc gaattagggt tagtaaaagg
     3001 gtcctaagga acagcgatat ctcccacccc atgagctgtc acggttttat ttacatgggg
     3061 tcaggattcc acgagggtag tgaaccattt tagtcacaag ggcagtggct gaagatcaag
     3121 gagcgggcag tgaactctcc tgaatcttcg cctgcttctt cattctcctt cgtttagcta
     3181 atagaataac tgctgagttg tgaacagtaa ggtgtatgtg aggtgctcga aaacaaggtt
     3241 tcaggtgacg cccccagaat aaaatttgga cggggggttc agtggtggca ttgtgctatg
     3301 acaccaatat aaccctcaca aaccccttgg gcaataaata ctagtgtagg aatgaaacat
     3361 tctgaatatc tttaacaata gaaatccatg gggtggggac aagccgtaaa gactggatgt
     3421 ccatctcaca cgaatttatg gctatgggca acacataatc ctagtgcaat atgatactgg
     3481 ggttattaag atgtgtccca ggcagggacc aagacaggtg aaccatgttg ttacactcta
     3541 tttgtaacaa ggggaaagag agtggacgcc gacagcagcg gactccactg gttgtctcta
     3601 acacccccga aaattaaacg gggctccacg ccaatggggc ccataaacaa agacaagtgg
     3661 ccactctttt ttttgaaatt gtggagtggg ggcacgcgtc agcccccaca cgccgccctg
     3721 cggttttgga ctgtaaaata agggtgtaat aacttggctg attgtaaccc cgctaaccac
     3781 tgcggtcaaa ccacttgccc acaaaaccac taatggcacc ccggggaata cctgcataag
     3841 taggtgggcg ggccaagata ggggcgcgat tgctgcgatc tggaggacaa attacacaca
     3901 cttgcgcctg agcgccaagc acagggttgt tggtcctcat attcacgagg tcgctgagag
     3961 cacggtgggc taatgttgcc atgggtagca tatactaccc aaatatctgg atagcatatg
     4021 ctatcctaat ctatatctgg gtagcatagg ctatcctaat ctatatctgg gtagcatatg
     4081 ctatcctaat ctatatctgg gtagtatatg ctatcctaat ttatatctgg gtagcatagg
     4141 ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagtatatg
     4201 ctatcctaat ctgtatccgg gtagcatatg ctatcctaat agagattagg gtagtatatg
     4261 ctatcctaat ttatatctgg gtagcatata ctacccaaat atctggatag catatgctat
     4321 cctaatctat atctgggtag catatgctat cctaatctat atctgggtag cataggctat
     4381 cctaatctat atctgggtag catatgctat cctaatctat atctgggtag tatatgctat
     4441 cctaatttat atctgggtag cataggctat cctaatctat atctgggtag catatgctat
     4501 cctaatctat atctgggtag tatatgctat cctaatctgt atccgggtag catatgctat
     4561 cctcatgata agctgtcaaa catgagaatt ttcttgaaga cgaaagggcc tcgtgatacg
     4621 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     4681 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     4741 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     4801 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     4861 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     4921 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     4981 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     5041 tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     5101 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     5161 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     5221 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     5281 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     5341 tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     5401 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     5461 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     5521 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     5581 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     5641 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     5701 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     5761 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     5821 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     5881 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     5941 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     6001 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     6061 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     6121 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     6181 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     6241 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     6301 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     6361 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     6421 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     6481 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     6541 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     6601 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     6661 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
     6721 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
     6781 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagctcta
     6841 gctagaggtc gaccaattct catgtttgac agcttatcat cgcagatccg ggcaacgttg
     6901 ttgccattgc tgcaggcgca gaactggtag gtatggaaga tctatacatt gaatcaatat
     6961 tggcaattag ccatattagt cattggttat atagcataaa tcaatattgg ctattggcca
     7021 ttgcatacgt tgtatctata tcataatatg tacatttata ttggctcatg tccaatatga
     7081 ccgccatgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
     7141 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
     7201 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     7261 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     7321 gcagtacatc aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa
     7381 tggcccgcct ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac
     7441 atctacgtat tagtcatcgc tattaccatg
//
