LOCUS       sequence-274525-        6654 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 6654)
  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 6654)
  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: D9_Ld_190920
FEATURES             Location/Qualifiers
     source          1..6654
                     /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             1432..1449
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1501..1556
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1555..1574)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3792..3810)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3878..3982
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3983..4051,4052..4843)
                     /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(4201..4220)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5014..5602
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5503..5522
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5756..5773
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5890..5911
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5926..5931,5932..5949,5950..5956)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5964..5980
                     /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     5969..5991
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5988..6004
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5988..6004
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        6275..6654
                     /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 gccaccagct attggagcga ggaagtgaac
      841 agagtccgga cctacgccgc cgtgaactac ctggaacgga tcgccaagca gcccaacgtg
      901 tccgccctgc ccagcggcct gctgttcacc atcgagcgga gaggcttcgg cgacagagcc
      961 cctgccgccg aggacaagtg cgagatgcac tacaccatcc actaccggtt ccccggcatc
     1021 gtggaaagca cccggcacca cccctacccc gtgcggagaa gccctagcca gctgatccct
     1081 ggcatggccg aggccatgca gctgatgcgc gagggcgacc ggtggtatct gcacgtgccc
     1141 taccagctgg gctacggcaa agagggctgc aaagaaaaga aggtgcccag cctgagcaac
     1201 ctgcgggtgg aaatcgagct gtataagtgc gagagcgcca gcggcaagac cagcgccgag
     1261 atcgacgcct acctggccaa gtacatgaag acccggatcc ccgagaaggc cgctcccgtg
     1321 gactacaccg acctgggcgc gcccggcagc ctgcaccaca tcctggacgc ccagaagatg
     1381 ctgtggaacc acagggacag gaacctgcct cctctggccc ctctgggccc ccaccaccat
     1441 caccaccatt gagtctagac acgtgattaa ttaaggatcc cccgacctcg acctctggct
     1501 aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg
     1561 gaaggacata tgggagggca aatcatttgg tcgagatccc tcggagatct ctagctagag
     1621 gatcgatccc cgccccggac gaactaaacc tgactacgac atctctgccc cttcttcgcg
     1681 gggcagtgca tgtaatccct tcagttggtt ggtacaactt gccaactggg ccctgttcca
     1741 catgtgacac ggggggggac caaacacaaa ggggttctct gactgtagtt gacatcctta
     1801 taaatggatg tgcacatttg ccaacactga gtggctttca tcctggagca gactttgcag
     1861 tctgtggact gcaacacaac attgccttta tgtgtaactc ttggctgaag ctcttacacc
     1921 aatgctgggg gacatgtacc tcccaggggc ccaggaagac tacgggaggc tacaccaacg
     1981 tcaatcagag gggcctgtgt agctaccgat aagcggaccc tcaagagggc attagcaata
     2041 gtgtttataa ggcccccttg ttaaccctaa acgggtagca tatgcttccc gggtagtagt
     2101 atatactatc cagactaacc ctaattcaat agcatatgtt acccaacggg aagcatatgc
     2161 tatcgaatta gggttagtaa aagggtccta aggaacagcg atatctccca ccccatgagc
     2221 tgtcacggtt ttatttacat ggggtcagga ttccacgagg gtagtgaacc attttagtca
     2281 caagggcagt ggctgaagat caaggagcgg gcagtgaact ctcctgaatc ttcgcctgct
     2341 tcttcattct ccttcgttta gctaatagaa taactgctga gttgtgaaca gtaaggtgta
     2401 tgtgaggtgc tcgaaaacaa ggtttcaggt gacgccccca gaataaaatt tggacggggg
     2461 gttcagtggt ggcattgtgc tatgacacca atataaccct cacaaacccc ttgggcaata
     2521 aatactagtg taggaatgaa acattctgaa tatctttaac aatagaaatc catggggtgg
     2581 ggacaagccg taaagactgg atgtccatct cacacgaatt tatggctatg ggcaacacat
     2641 aatcctagtg caatatgata ctggggttat taagatgtgt cccaggcagg gaccaagaca
     2701 ggtgaaccat gttgttacac tctatttgta acaaggggaa agagagtgga cgccgacagc
     2761 agcggactcc actggttgtc tctaacaccc ccgaaaatta aacggggctc cacgccaatg
     2821 gggcccataa acaaagacaa gtggccactc ttttttttga aattgtggag tgggggcacg
     2881 cgtcagcccc cacacgccgc cctgcggttt tggactgtaa aataagggtg taataacttg
     2941 gctgattgta accccgctaa ccactgcggt caaaccactt gcccacaaaa ccactaatgg
     3001 caccccgggg aatacctgca taagtaggtg ggcgggccaa gataggggcg cgattgctgc
     3061 gatctggagg acaaattaca cacacttgcg cctgagcgcc aagcacaggg ttgttggtcc
     3121 tcatattcac gaggtcgctg agagcacggt gggctaatgt tgccatgggt agcatatact
     3181 acccaaatat ctggatagca tatgctatcc taatctatat ctgggtagca taggctatcc
     3241 taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagta tatgctatcc
     3301 taatttatat ctgggtagca taggctatcc taatctatat ctgggtagca tatgctatcc
     3361 taatctatat ctgggtagta tatgctatcc taatctgtat ccgggtagca tatgctatcc
     3421 taatagagat tagggtagta tatgctatcc taatttatat ctgggtagca tatactaccc
     3481 aaatatctgg atagcatatg ctatcctaat ctatatctgg gtagcatatg ctatcctaat
     3541 ctatatctgg gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat
     3601 ctatatctgg gtagtatatg ctatcctaat ttatatctgg gtagcatagg ctatcctaat
     3661 ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat
     3721 ctgtatccgg gtagcatatg ctatcctcat gataagctgt caaacatgag aattttcttg
     3781 aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     3841 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3901 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     3961 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     4021 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     4081 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     4141 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     4201 gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat
     4261 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     4321 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     4381 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     4441 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     4501 cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac
     4561 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     4621 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     4681 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     4741 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     4801 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     4861 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     4921 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4981 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5041 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5101 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5161 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5221 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5281 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5341 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5401 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5461 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     5521 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5581 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5641 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     5701 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     5761 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     5821 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     5881 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     5941 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
     6001 tgaccatgat tacgccaagc tctagctaga ggtcgaccaa ttctcatgtt tgacagctta
     6061 tcatcgcaga tccgggcaac gttgttgcca ttgctgcagg cgcagaactg gtaggtatgg
     6121 aagatctata cattgaatca atattggcaa ttagccatat tagtcattgg ttatatagca
     6181 taaatcaata ttggctattg gccattgcat acgttgtatc tatatcataa tatgtacatt
     6241 tatattggct catgtccaat atgaccgcca tgttgacatt gattattgac tagttattaa
     6301 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     6361 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     6421 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     6481 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtccgccc
     6541 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     6601 cgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catg
//
