LOCUS       sequence-274895-        7434 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 7434)
  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 7434)
  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: D41_Ld_180450
FEATURES             Location/Qualifiers
     source          1..7434
                     /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             2592..2609
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2661..2716
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2715..2734)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4952..4970)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5038..5142
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5143..5211,5212..6003)
                     /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(5361..5380)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6174..6762
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6663..6682
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6916..6933
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7050..7071
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7086..7091,7092..7109,7110..7116)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7124..7140
                     /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     7129..7151
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7148..7164
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7148..7164
                     /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 gccacctcta
     1201 cacctcacca cggctacgcc agctgcgacc ccagcgtggt gcagagcagc ggctacatca
     1261 acatccccgg cgtgaaccag accctgaagc actacttcta ctggctgttc ggccccagaa
     1321 agtggcccaa cgacttccgc gagccccccg tgatcatgtg gatgacaggc ggccctggct
     1381 gcagcagctc catggccctg ctgaccgagc tgggcccctg catgatgaac gagacaagcg
     1441 gcgagctgta ccacaacacc tacggctgga acgacgaggc ctacctgctg ttcgtggacc
     1501 agcccaccgg cgtgggctac agctacggcg acaagagcaa ctacgcccac aaccagagcg
     1561 aggtggccga ggatatgtac aacttcctgc agctgttcgc cagacggttc accagcccca
     1621 gcatcaccgg cgccaacgat ttctacatca tcggcgagtc ctacggcggc cattacgtgc
     1681 ccgccgtgtc ccacagaatc ctgatgggca acgagcggag cgacggcctg cacatcaacc
     1741 tgaagggaat cgccatcggc aacggcctga ccgaccccta cacccagctg cccttccacg
     1801 cccagaccgc ctactactgg tgcaaagaga agctgggcgc tccctgtatc accgagaagg
     1861 cctacgagga aatgctgagc ctgctgcccg cctgcctgga aaagaccaag aagtgcaacg
     1921 agggccccga cgacagcgac gtgtcctgta gcgtggccac agccctgtgg gccgagtatg
     1981 tggaccacta ctacgccacc ggccggaact cctacgacat ccggaagcag tgtatcggcg
     2041 acctgtgcta ccccatgcag aacaccatcg acttctacca caagcctacc gtgcgggcca
     2101 gcctgggagc ctctgccaag gcccagtggt ccacctgtaa cggcgaggtg tccgccctgt
     2161 tcgagcggga ctacatgcgg aacttcaact tcaccttccc ccacatgctg gacatgggca
     2221 tccgggtgct gatctacgcc ggcgacatgg acttcatctg caactggctg ggcaatgagg
     2281 cctgggtcaa ggccctgcag tggttcggca ccgacggctt caacaccgcc cccaacgtgg
     2341 aatttgccgt gtctggcaga tgggccggac aggaaagaag ctacgggggc ctgagcttcg
     2401 tgcgcatcta cgacgccgga cacatggtgc ccatggatca gcctgaggtg gcactgttca
     2461 tggtgcaccg gtttctgcac gaccggcggc tggctggcgc gcccggcagc ctgcaccaca
     2521 tcctggacgc ccagaagatg ctgtggaacc acagggacag gaacctgcct cctctggccc
     2581 ctctgggccc ccaccaccat caccaccatt gagtctagac acgtgattaa ttaaggatcc
     2641 cccgacctcg acctctggct aataaaggaa atttattttc attgcaatag tgtgttggaa
     2701 ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttgg tcgagatccc
     2761 tcggagatct ctagctagag gatcgatccc cgccccggac gaactaaacc tgactacgac
     2821 atctctgccc cttcttcgcg gggcagtgca tgtaatccct tcagttggtt ggtacaactt
     2881 gccaactggg ccctgttcca catgtgacac ggggggggac caaacacaaa ggggttctct
     2941 gactgtagtt gacatcctta taaatggatg tgcacatttg ccaacactga gtggctttca
     3001 tcctggagca gactttgcag tctgtggact gcaacacaac attgccttta tgtgtaactc
     3061 ttggctgaag ctcttacacc aatgctgggg gacatgtacc tcccaggggc ccaggaagac
     3121 tacgggaggc tacaccaacg tcaatcagag gggcctgtgt agctaccgat aagcggaccc
     3181 tcaagagggc attagcaata gtgtttataa ggcccccttg ttaaccctaa acgggtagca
     3241 tatgcttccc gggtagtagt atatactatc cagactaacc ctaattcaat agcatatgtt
     3301 acccaacggg aagcatatgc tatcgaatta gggttagtaa aagggtccta aggaacagcg
     3361 atatctccca ccccatgagc tgtcacggtt ttatttacat ggggtcagga ttccacgagg
     3421 gtagtgaacc attttagtca caagggcagt ggctgaagat caaggagcgg gcagtgaact
     3481 ctcctgaatc ttcgcctgct tcttcattct ccttcgttta gctaatagaa taactgctga
     3541 gttgtgaaca gtaaggtgta tgtgaggtgc tcgaaaacaa ggtttcaggt gacgccccca
     3601 gaataaaatt tggacggggg gttcagtggt ggcattgtgc tatgacacca atataaccct
     3661 cacaaacccc ttgggcaata aatactagtg taggaatgaa acattctgaa tatctttaac
     3721 aatagaaatc catggggtgg ggacaagccg taaagactgg atgtccatct cacacgaatt
     3781 tatggctatg ggcaacacat aatcctagtg caatatgata ctggggttat taagatgtgt
     3841 cccaggcagg gaccaagaca ggtgaaccat gttgttacac tctatttgta acaaggggaa
     3901 agagagtgga cgccgacagc agcggactcc actggttgtc tctaacaccc ccgaaaatta
     3961 aacggggctc cacgccaatg gggcccataa acaaagacaa gtggccactc ttttttttga
     4021 aattgtggag tgggggcacg cgtcagcccc cacacgccgc cctgcggttt tggactgtaa
     4081 aataagggtg taataacttg gctgattgta accccgctaa ccactgcggt caaaccactt
     4141 gcccacaaaa ccactaatgg caccccgggg aatacctgca taagtaggtg ggcgggccaa
     4201 gataggggcg cgattgctgc gatctggagg acaaattaca cacacttgcg cctgagcgcc
     4261 aagcacaggg ttgttggtcc tcatattcac gaggtcgctg agagcacggt gggctaatgt
     4321 tgccatgggt agcatatact acccaaatat ctggatagca tatgctatcc taatctatat
     4381 ctgggtagca taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat
     4441 ctgggtagta tatgctatcc taatttatat ctgggtagca taggctatcc taatctatat
     4501 ctgggtagca tatgctatcc taatctatat ctgggtagta tatgctatcc taatctgtat
     4561 ccgggtagca tatgctatcc taatagagat tagggtagta tatgctatcc taatttatat
     4621 ctgggtagca tatactaccc aaatatctgg atagcatatg ctatcctaat ctatatctgg
     4681 gtagcatatg ctatcctaat ctatatctgg gtagcatagg ctatcctaat ctatatctgg
     4741 gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat ttatatctgg
     4801 gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg
     4861 gtagtatatg ctatcctaat ctgtatccgg gtagcatatg ctatcctcat gataagctgt
     4921 caaacatgag aattttcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt
     4981 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     5041 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     5101 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     5161 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     5221 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     5281 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     5341 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa
     5401 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     5461 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     5521 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     5581 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     5641 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca
     5701 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     5761 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     5821 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     5881 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     5941 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     6001 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     6061 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     6121 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     6181 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     6241 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     6301 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     6361 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     6421 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     6481 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     6541 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     6601 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     6661 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6721 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     6781 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     6841 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     6901 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     6961 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     7021 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     7081 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     7141 tttcacacag gaaacagcta tgaccatgat tacgccaagc tctagctaga ggtcgaccaa
     7201 ttctcatgtt tgacagctta tcatcgcaga tccgggcaac gttgttgcca ttgctgcagg
     7261 cgcagaactg gtaggtatgg aagatctata cattgaatca atattggcaa ttagccatat
     7321 tagtcattgg ttatatagca taaatcaata ttggctattg gccattgcat acgttgtatc
     7381 tatatcataa tatgtacatt tatattggct catgtccaat atgaccgcca tgtt
//
