LOCUS       sequence-262324-        6978 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 6978)
  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 6978)
  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: D29_Ld_251870
FEATURES             Location/Qualifiers
     source          1..6978
                     /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             2142..2159
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2211..2266
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2265..2284)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4502..4520)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4588..4692
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4693..4761,4762..5553)
                     /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(4911..4930)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5724..6312
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6213..6232
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6466..6483
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6600..6621
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6636..6641,6642..6659,6660..6666)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6674..6690
                     /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     6679..6701
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6698..6714
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6698..6714
                     /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 ccgccctgta ctactacagc gtgtacgact acgagaagca catgagcatg gtgcagcgga
     1261 agtacagcgg ctacgacccc ctgaccgact gcgccacccc ttttggccag ctgctgggcg
     1321 tggccgacga tgtgcccgcc tacagcaact gcaacaccaa gttcagcagc acctacatca
     1381 actacgtgtc cctgatggac cccatggaca acggcagacg gggcgacccc agcgagacac
     1441 ggatcgtgat gaccgcctac cggtacaccg ccttcgacta ctgcatgcgg tggctcgtgt
     1501 ggaaccgggg cgtgatgccc cggctggtgg aaaacaccaa ccagctctgg aaaaccgtgg
     1561 actacttcaa ccccgccaag cccgagcagg ggtggtccgc cgagtacatc accaactacg
     1621 aagaggtcac aggcatggaa gagagaaagt tcaacgcccc cagacgcgcc gacgccatcg
     1681 tgtaccggat ggacaagaac accatccctg ccggccacat tgccgtggtg gtcaaggtgg
     1741 aagatgacgt ggaagccgct ggcggccctg agaagctgaa agaactgaag aagatgcggc
     1801 tgcaccccag gcgggtgtac gtggcagagc agaattggaa gaaccagccc tggggcggac
     1861 acaactacag ccgggtgctg cagttcaagt ggcgcgctgt ggccgagaag gcctacgagg
     1921 gctgctacgt ggaccccgac ggcctgggca tcatcggcgt cgtcagagtg ggcaaggcca
     1981 tgcccctgag agccgcccca gacccctacc aggaagccct ggacatggat aacgacggcg
     2041 acctgggcgc gcccggcagc ctgcaccaca tcctggacgc ccagaagatg ctgtggaacc
     2101 acagggacag gaacctgcct cctctggccc ctctgggccc ccaccaccat caccaccatt
     2161 gagtctagac acgtgattaa ttaaggatcc cccgacctcg acctctggct aataaaggaa
     2221 atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata
     2281 tgggagggca aatcatttgg tcgagatccc tcggagatct ctagctagag gatcgatccc
     2341 cgccccggac gaactaaacc tgactacgac atctctgccc cttcttcgcg gggcagtgca
     2401 tgtaatccct tcagttggtt ggtacaactt gccaactggg ccctgttcca catgtgacac
     2461 ggggggggac caaacacaaa ggggttctct gactgtagtt gacatcctta taaatggatg
     2521 tgcacatttg ccaacactga gtggctttca tcctggagca gactttgcag tctgtggact
     2581 gcaacacaac attgccttta tgtgtaactc ttggctgaag ctcttacacc aatgctgggg
     2641 gacatgtacc tcccaggggc ccaggaagac tacgggaggc tacaccaacg tcaatcagag
     2701 gggcctgtgt agctaccgat aagcggaccc tcaagagggc attagcaata gtgtttataa
     2761 ggcccccttg ttaaccctaa acgggtagca tatgcttccc gggtagtagt atatactatc
     2821 cagactaacc ctaattcaat agcatatgtt acccaacggg aagcatatgc tatcgaatta
     2881 gggttagtaa aagggtccta aggaacagcg atatctccca ccccatgagc tgtcacggtt
     2941 ttatttacat ggggtcagga ttccacgagg gtagtgaacc attttagtca caagggcagt
     3001 ggctgaagat caaggagcgg gcagtgaact ctcctgaatc ttcgcctgct tcttcattct
     3061 ccttcgttta gctaatagaa taactgctga gttgtgaaca gtaaggtgta tgtgaggtgc
     3121 tcgaaaacaa ggtttcaggt gacgccccca gaataaaatt tggacggggg gttcagtggt
     3181 ggcattgtgc tatgacacca atataaccct cacaaacccc ttgggcaata aatactagtg
     3241 taggaatgaa acattctgaa tatctttaac aatagaaatc catggggtgg ggacaagccg
     3301 taaagactgg atgtccatct cacacgaatt tatggctatg ggcaacacat aatcctagtg
     3361 caatatgata ctggggttat taagatgtgt cccaggcagg gaccaagaca ggtgaaccat
     3421 gttgttacac tctatttgta acaaggggaa agagagtgga cgccgacagc agcggactcc
     3481 actggttgtc tctaacaccc ccgaaaatta aacggggctc cacgccaatg gggcccataa
     3541 acaaagacaa gtggccactc ttttttttga aattgtggag tgggggcacg cgtcagcccc
     3601 cacacgccgc cctgcggttt tggactgtaa aataagggtg taataacttg gctgattgta
     3661 accccgctaa ccactgcggt caaaccactt gcccacaaaa ccactaatgg caccccgggg
     3721 aatacctgca taagtaggtg ggcgggccaa gataggggcg cgattgctgc gatctggagg
     3781 acaaattaca cacacttgcg cctgagcgcc aagcacaggg ttgttggtcc tcatattcac
     3841 gaggtcgctg agagcacggt gggctaatgt tgccatgggt agcatatact acccaaatat
     3901 ctggatagca tatgctatcc taatctatat ctgggtagca taggctatcc taatctatat
     3961 ctgggtagca tatgctatcc taatctatat ctgggtagta tatgctatcc taatttatat
     4021 ctgggtagca taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat
     4081 ctgggtagta tatgctatcc taatctgtat ccgggtagca tatgctatcc taatagagat
     4141 tagggtagta tatgctatcc taatttatat ctgggtagca tatactaccc aaatatctgg
     4201 atagcatatg ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg
     4261 gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg
     4321 gtagtatatg ctatcctaat ttatatctgg gtagcatagg ctatcctaat ctatatctgg
     4381 gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat ctgtatccgg
     4441 gtagcatatg ctatcctcat gataagctgt caaacatgag aattttcttg aagacgaaag
     4501 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     4561 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     4621 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     4681 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     4741 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     4801 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     4861 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     4921 gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     4981 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     5041 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     5101 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     5161 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     5221 gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     5281 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     5341 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     5401 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     5461 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     5521 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     5581 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     5641 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     5701 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     5761 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     5821 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     5881 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     5941 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6001 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     6061 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     6121 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6181 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6241 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     6301 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     6361 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     6421 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     6481 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     6541 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     6601 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     6661 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     6721 tacgccaagc tctagctaga ggtcgaccaa ttctcatgtt tgacagctta tcatcgcaga
     6781 tccgggcaac gttgttgcca ttgctgcagg cgcagaactg gtaggtatgg aagatctata
     6841 cattgaatca atattggcaa ttagccatat tagtcattgg ttatatagca taaatcaata
     6901 ttggctattg gccattgcat acgttgtatc tatatcataa tatgtacatt tatattggct
     6961 catgtccaat atgaccgc
//
