LOCUS       sequence-262297-        7098 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 7098)
  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 7098)
  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: D2_Ld_030320
FEATURES             Location/Qualifiers
     source          1..7098
                     /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             2262..2279
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2331..2386
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2385..2404)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4622..4640)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4708..4812
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4813..4881,4882..5673)
                     /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(5031..5050)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5844..6432
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6333..6352
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6586..6603
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6720..6741
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6756..6761,6762..6779,6780..6786)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6794..6810
                     /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     6799..6821
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6818..6834
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6818..6834
                     /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 ccagatggat cagccagaga cacgccgacg acagcggcgt gggagataag gccctggccc
     1261 acaagggccg gctgagactg agccagaatg ccagcggaag cgccgtgcgg cggagaggca
     1321 gcaaggacga gaacggcagc gaccaggtcg gagtggacga cggcaatggc ggagccgtgg
     1381 gcagaagaag aagaggcgct ggcggcgacc tgcggagaag aaggcctcag gaaagaggcg
     1441 ccaaccccca ccacggcggc agcgacggcg acgagaacag ctacgacgac gacgataccg
     1501 gcggcatccc cgagacagac gccaacggcg tgaagctgac ccggctgcag cggaagaagc
     1561 tggccaaaga gcgcgagcgg gaagagaggc ggcaggccca ggaagccgcc ctggaagccc
     1621 agagacagcg gaccgacgcc aacgagctga gagatgccga ggccgccaga cgggaagagg
     1681 aactgaaggc cgccgaagag gccgccctgc agcagctgcg gcgggacaag aagaaagccg
     1741 aggacgagga atacgccaaa tgggtgtccc acatcggcgt ggaagagaga ggcgagctgg
     1801 gcgacgagga aagagagcgg cagaccagac tgcagagctt cctgctgggc agagccgccc
     1861 aggtgcaggc cagagcccag agatctgccg ctggcgccaa gagcagcccc agcgaagagg
     1921 aatgcatcca cgtgctgatc ctgcagaccg ccgccaggga cctgaaggtg tccgtggaag
     1981 aactggtgtc caccatcgag cagctgagcc aggccgacaa ggtgcacggc gtgttcgacg
     2041 accggggcaa atacgtgttc attgcccccg agcagttccc cctgctggcc cagttcatcc
     2101 ggctgcgggg cagagtgtcc gtgcaggaat tcacccgcga gtgcaaccgg atcgtgatgc
     2161 agtctggcgc gcccggcagc ctgcaccaca tcctggacgc ccagaagatg ctgtggaacc
     2221 acagggacag gaacctgcct cctctggccc ctctgggccc ccaccaccat caccaccatt
     2281 gagtctagac acgtgattaa ttaaggatcc cccgacctcg acctctggct aataaaggaa
     2341 atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata
     2401 tgggagggca aatcatttgg tcgagatccc tcggagatct ctagctagag gatcgatccc
     2461 cgccccggac gaactaaacc tgactacgac atctctgccc cttcttcgcg gggcagtgca
     2521 tgtaatccct tcagttggtt ggtacaactt gccaactggg ccctgttcca catgtgacac
     2581 ggggggggac caaacacaaa ggggttctct gactgtagtt gacatcctta taaatggatg
     2641 tgcacatttg ccaacactga gtggctttca tcctggagca gactttgcag tctgtggact
     2701 gcaacacaac attgccttta tgtgtaactc ttggctgaag ctcttacacc aatgctgggg
     2761 gacatgtacc tcccaggggc ccaggaagac tacgggaggc tacaccaacg tcaatcagag
     2821 gggcctgtgt agctaccgat aagcggaccc tcaagagggc attagcaata gtgtttataa
     2881 ggcccccttg ttaaccctaa acgggtagca tatgcttccc gggtagtagt atatactatc
     2941 cagactaacc ctaattcaat agcatatgtt acccaacggg aagcatatgc tatcgaatta
     3001 gggttagtaa aagggtccta aggaacagcg atatctccca ccccatgagc tgtcacggtt
     3061 ttatttacat ggggtcagga ttccacgagg gtagtgaacc attttagtca caagggcagt
     3121 ggctgaagat caaggagcgg gcagtgaact ctcctgaatc ttcgcctgct tcttcattct
     3181 ccttcgttta gctaatagaa taactgctga gttgtgaaca gtaaggtgta tgtgaggtgc
     3241 tcgaaaacaa ggtttcaggt gacgccccca gaataaaatt tggacggggg gttcagtggt
     3301 ggcattgtgc tatgacacca atataaccct cacaaacccc ttgggcaata aatactagtg
     3361 taggaatgaa acattctgaa tatctttaac aatagaaatc catggggtgg ggacaagccg
     3421 taaagactgg atgtccatct cacacgaatt tatggctatg ggcaacacat aatcctagtg
     3481 caatatgata ctggggttat taagatgtgt cccaggcagg gaccaagaca ggtgaaccat
     3541 gttgttacac tctatttgta acaaggggaa agagagtgga cgccgacagc agcggactcc
     3601 actggttgtc tctaacaccc ccgaaaatta aacggggctc cacgccaatg gggcccataa
     3661 acaaagacaa gtggccactc ttttttttga aattgtggag tgggggcacg cgtcagcccc
     3721 cacacgccgc cctgcggttt tggactgtaa aataagggtg taataacttg gctgattgta
     3781 accccgctaa ccactgcggt caaaccactt gcccacaaaa ccactaatgg caccccgggg
     3841 aatacctgca taagtaggtg ggcgggccaa gataggggcg cgattgctgc gatctggagg
     3901 acaaattaca cacacttgcg cctgagcgcc aagcacaggg ttgttggtcc tcatattcac
     3961 gaggtcgctg agagcacggt gggctaatgt tgccatgggt agcatatact acccaaatat
     4021 ctggatagca tatgctatcc taatctatat ctgggtagca taggctatcc taatctatat
     4081 ctgggtagca tatgctatcc taatctatat ctgggtagta tatgctatcc taatttatat
     4141 ctgggtagca taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat
     4201 ctgggtagta tatgctatcc taatctgtat ccgggtagca tatgctatcc taatagagat
     4261 tagggtagta tatgctatcc taatttatat ctgggtagca tatactaccc aaatatctgg
     4321 atagcatatg ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg
     4381 gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg
     4441 gtagtatatg ctatcctaat ttatatctgg gtagcatagg ctatcctaat ctatatctgg
     4501 gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat ctgtatccgg
     4561 gtagcatatg ctatcctcat gataagctgt caaacatgag aattttcttg aagacgaaag
     4621 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     4681 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     4741 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     4801 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     4861 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     4921 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     4981 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     5041 gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     5101 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     5161 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     5221 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     5281 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     5341 gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     5401 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     5461 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     5521 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     5581 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     5641 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     5701 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     5761 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     5821 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     5881 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     5941 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     6001 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     6061 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6121 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     6181 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     6241 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6301 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6361 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     6421 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     6481 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     6541 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     6601 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     6661 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     6721 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     6781 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     6841 tacgccaagc tctagctaga ggtcgaccaa ttctcatgtt tgacagctta tcatcgcaga
     6901 tccgggcaac gttgttgcca ttgctgcagg cgcagaactg gtaggtatgg aagatctata
     6961 cattgaatca atattggcaa ttagccatat tagtcattgg ttatatagca taaatcaata
     7021 ttggctattg gccattgcat acgttgtatc tatatcataa tatgtacatt tatattggct
     7081 catgtccaat atgaccgc
//
