LOCUS       sequence-274558-        7734 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 7734)
  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 7734)
  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: D36_Ld_323600
FEATURES             Location/Qualifiers
     source          1..7734
                     /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             2512..2529
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2581..2636
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2635..2654)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4872..4890)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4958..5062
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5063..5131,5132..5923)
                     /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(5281..5300)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6094..6682
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6583..6602
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6836..6853
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6970..6991
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7006..7011,7012..7029,7030..7036)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7044..7060
                     /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     7049..7071
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7068..7084
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7068..7084
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7355..7734
                     /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 gccacctctg gatctgccac cgtgggagcc
      841 cacaacgaca gactgcccca gcccagctgg tccagaacct gcaccagccc ctacgaagtg
      901 tgcagcacca tcgaggtgta ccagtactgg gaggaaaaga agaaccggct gagcttcacc
      961 gtggacgcca tgatgaagga ccccaacgcc aagaacgtgc tgaccgtgtg gatggtggtg
     1021 gacgacgccg acgtgaacat ggccaagggc atcttccgcg agggcatcaa cgtggtcatc
     1081 gtgcacccca gaggcagcaa gcagacccag agcttcatga cctgcgccaa caccagcaac
     1141 ccctacaacc ggcagtgctg gatgagcgag gcctgtgccg ccgaactggc cgctgccgcc
     1201 agcagcacct ccaaccccct gaacatgtcc cactacagcg ccaagcaggc cgcccacgac
     1261 ctggactggg ctctgagaac actgggcgac ggcagaccta acgtggtgct gggccagggc
     1321 ctgagcagca tgctggtgct gcggctgctg cagtaccacc aggacaccaa ggccgccgtg
     1381 atcctgctgg actatgtgca ccctgtgatg ttcgacgtgt acaactactt cggaggcggc
     1441 agcatggtca ccgccctgca gcatctgctg gccatgtgcg acgaccaggc cgcttgcgtg
     1501 ggcagactgg gagccgccga gggcagctgg aacagactga tgacagtgat ccatatggcc
     1561 aaagagggca agctgagctg cgcctccaga ctgagatggg cccccagaag aggcggaaac
     1621 gccgccttcg ccgacgagct gagaggcgtg ctggccctga tgctgagata ccccgtgtac
     1681 cccttcctgg cctccaacgc cgctctgctg aacctgatcc ccagcctgct gtaccgggtg
     1741 cagcggtgca acaagaaaga cgtggccgca ctgaacaagc tgtacgacta cctgagcgcc
     1801 gccaaaaact acgagtgccc cgagagcgtg gccctgcaca tgcactggct ggtcaacgag
     1861 ttcctgcagg ctgccccccc tgccaacatc cacagcttca gaagcagcgc cgcagccaag
     1921 cacctgattc tgcccagctt ccacgccatg tccgccttcc acagcgctgc caagaagttc
     1981 ccccggatca gcagatccgc cgagtccaag cagctgcccg ccaatgccac ccagcctatt
     2041 ctgctgctga atgccgacac cgacgccctg ctgcctggcg gagctgcttc tcaggtggca
     2101 ctggccttca gacagctggg cgatagcgtg cagctgaggc agctgcgggg catcatgaac
     2161 aggcccagcg ctgtgctgac accctgcctc gtgaacaacc tgaagatgtt ccgggaccac
     2221 ggcgtgtggg ccgatgagga ccagtgtacc ctggatgtgg cccacaagat cgacttcatc
     2281 aacggcgcca ccgacatcta ctacggcacc agcgacgcct gggacttcga caagcccaac
     2341 agcgaggaaa caaacggcgg cagggacgac aacgacgacg agggcggctt ccacttcaag
     2401 tggcggttta tcgctggcgc gcccggcagc ctgcaccaca tcctggacgc ccagaagatg
     2461 ctgtggaacc acagggacag gaacctgcct cctctggccc ctctgggccc ccaccaccat
     2521 caccaccatt gagtctagac acgtgattaa ttaaggatcc cccgacctcg acctctggct
     2581 aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg
     2641 gaaggacata tgggagggca aatcatttgg tcgagatccc tcggagatct ctagctagag
     2701 gatcgatccc cgccccggac gaactaaacc tgactacgac atctctgccc cttcttcgcg
     2761 gggcagtgca tgtaatccct tcagttggtt ggtacaactt gccaactggg ccctgttcca
     2821 catgtgacac ggggggggac caaacacaaa ggggttctct gactgtagtt gacatcctta
     2881 taaatggatg tgcacatttg ccaacactga gtggctttca tcctggagca gactttgcag
     2941 tctgtggact gcaacacaac attgccttta tgtgtaactc ttggctgaag ctcttacacc
     3001 aatgctgggg gacatgtacc tcccaggggc ccaggaagac tacgggaggc tacaccaacg
     3061 tcaatcagag gggcctgtgt agctaccgat aagcggaccc tcaagagggc attagcaata
     3121 gtgtttataa ggcccccttg ttaaccctaa acgggtagca tatgcttccc gggtagtagt
     3181 atatactatc cagactaacc ctaattcaat agcatatgtt acccaacggg aagcatatgc
     3241 tatcgaatta gggttagtaa aagggtccta aggaacagcg atatctccca ccccatgagc
     3301 tgtcacggtt ttatttacat ggggtcagga ttccacgagg gtagtgaacc attttagtca
     3361 caagggcagt ggctgaagat caaggagcgg gcagtgaact ctcctgaatc ttcgcctgct
     3421 tcttcattct ccttcgttta gctaatagaa taactgctga gttgtgaaca gtaaggtgta
     3481 tgtgaggtgc tcgaaaacaa ggtttcaggt gacgccccca gaataaaatt tggacggggg
     3541 gttcagtggt ggcattgtgc tatgacacca atataaccct cacaaacccc ttgggcaata
     3601 aatactagtg taggaatgaa acattctgaa tatctttaac aatagaaatc catggggtgg
     3661 ggacaagccg taaagactgg atgtccatct cacacgaatt tatggctatg ggcaacacat
     3721 aatcctagtg caatatgata ctggggttat taagatgtgt cccaggcagg gaccaagaca
     3781 ggtgaaccat gttgttacac tctatttgta acaaggggaa agagagtgga cgccgacagc
     3841 agcggactcc actggttgtc tctaacaccc ccgaaaatta aacggggctc cacgccaatg
     3901 gggcccataa acaaagacaa gtggccactc ttttttttga aattgtggag tgggggcacg
     3961 cgtcagcccc cacacgccgc cctgcggttt tggactgtaa aataagggtg taataacttg
     4021 gctgattgta accccgctaa ccactgcggt caaaccactt gcccacaaaa ccactaatgg
     4081 caccccgggg aatacctgca taagtaggtg ggcgggccaa gataggggcg cgattgctgc
     4141 gatctggagg acaaattaca cacacttgcg cctgagcgcc aagcacaggg ttgttggtcc
     4201 tcatattcac gaggtcgctg agagcacggt gggctaatgt tgccatgggt agcatatact
     4261 acccaaatat ctggatagca tatgctatcc taatctatat ctgggtagca taggctatcc
     4321 taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagta tatgctatcc
     4381 taatttatat ctgggtagca taggctatcc taatctatat ctgggtagca tatgctatcc
     4441 taatctatat ctgggtagta tatgctatcc taatctgtat ccgggtagca tatgctatcc
     4501 taatagagat tagggtagta tatgctatcc taatttatat ctgggtagca tatactaccc
     4561 aaatatctgg atagcatatg ctatcctaat ctatatctgg gtagcatatg ctatcctaat
     4621 ctatatctgg gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat
     4681 ctatatctgg gtagtatatg ctatcctaat ttatatctgg gtagcatagg ctatcctaat
     4741 ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat
     4801 ctgtatccgg gtagcatatg ctatcctcat gataagctgt caaacatgag aattttcttg
     4861 aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     4921 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     4981 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     5041 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     5101 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     5161 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     5221 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     5281 gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat
     5341 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     5401 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     5461 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     5521 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     5581 cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac
     5641 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     5701 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     5761 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     5821 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     5881 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     5941 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     6001 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     6061 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     6121 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     6181 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     6241 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     6301 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     6361 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     6421 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     6481 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6541 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6601 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6661 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     6721 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     6781 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     6841 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     6901 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     6961 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     7021 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
     7081 tgaccatgat tacgccaagc tctagctaga ggtcgaccaa ttctcatgtt tgacagctta
     7141 tcatcgcaga tccgggcaac gttgttgcca ttgctgcagg cgcagaactg gtaggtatgg
     7201 aagatctata cattgaatca atattggcaa ttagccatat tagtcattgg ttatatagca
     7261 taaatcaata ttggctattg gccattgcat acgttgtatc tatatcataa tatgtacatt
     7321 tatattggct catgtccaat atgaccgcca tgttgacatt gattattgac tagttattaa
     7381 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     7441 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     7501 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     7561 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtccgccc
     7621 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     7681 cgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catg
//
