LOCUS       sequence-262381-        6855 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 6855)
  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 6855)
  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: D89_Ld_343450
FEATURES             Location/Qualifiers
     source          1..6855
                     /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     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             2019..2036
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2088..2143
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2142..2161)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4379..4397)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4465..4569
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4570..4638,4639..5430)
                     /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(4788..4807)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5601..6189
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6090..6109
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6343..6360
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6477..6498
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6513..6518,6519..6536,6537..6543)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6551..6567
                     /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     6556..6578
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6575..6591
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6575..6591
                     /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 gcggccgccg
     1201 gaattacagg ttattctact ggtacgatca tttctcccca ggctaacgca tatcggagtc
     1261 ggcgcacact gagcccaatg gattattaca aactgcctgt atgtcagccg tctgacgagg
     1321 tcatgaaggc aaggcgtgag caccccctca ttggggatat tctgactggg aaccgtctgg
     1381 tgcccacaat gtttgaattc agggtcggcg aggatgtaaa gtgtgccacc ctctgcgatg
     1441 cctctttcac cgtgaaagcc gtgcggcgcg cgaactacat gattaataat gactactatg
     1501 tgcggatgtt cctggataac aaaccgctcg tctctgcatc tccccacgag ggcagcaacg
     1561 cgtacctgct tggttatcca cttggcgccc agaatgacgt tgagaaaacc caggtgaaaa
     1621 ccaatatcat tcacaatcac cttgacttca caataagaat taagaatcgc gcgatttctc
     1681 agtttaccgg cgaagaggtg gtgggtttta aggtcgtggc taggtcactc gctgaagtgg
     1741 gtacctgcac ggctaccgct ttccagcact cttccaggcc ctacatcctt ccaagctacc
     1801 gcgacgggaa ggacctgaaa gtgcccttta cctacagcgt gacctgggag cgcagcaacg
     1861 aggagtatcc aatcgagcac cgcatcggag aggacacgca gaggcgcggc cataagatcg
     1921 ccggcgcgcc cggcagcctg caccacatcc tggacgccca gaagatgctg tggaaccaca
     1981 gggacaggaa cctgcctcct ctggcccctc tgggccccca ccaccatcac caccattgag
     2041 tctagacacg tgattaatta aggatccccc gacctcgacc tctggctaat aaaggaaatt
     2101 tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg
     2161 gagggcaaat catttggtcg agatccctcg gagatctcta gctagaggat cgatccccgc
     2221 cccggacgaa ctaaacctga ctacgacatc tctgcccctt cttcgcgggg cagtgcatgt
     2281 aatcccttca gttggttggt acaacttgcc aactgggccc tgttccacat gtgacacggg
     2341 gggggaccaa acacaaaggg gttctctgac tgtagttgac atccttataa atggatgtgc
     2401 acatttgcca acactgagtg gctttcatcc tggagcagac tttgcagtct gtggactgca
     2461 acacaacatt gcctttatgt gtaactcttg gctgaagctc ttacaccaat gctgggggac
     2521 atgtacctcc caggggccca ggaagactac gggaggctac accaacgtca atcagagggg
     2581 cctgtgtagc taccgataag cggaccctca agagggcatt agcaatagtg tttataaggc
     2641 ccccttgtta accctaaacg ggtagcatat gcttcccggg tagtagtata tactatccag
     2701 actaacccta attcaatagc atatgttacc caacgggaag catatgctat cgaattaggg
     2761 ttagtaaaag ggtcctaagg aacagcgata tctcccaccc catgagctgt cacggtttta
     2821 tttacatggg gtcaggattc cacgagggta gtgaaccatt ttagtcacaa gggcagtggc
     2881 tgaagatcaa ggagcgggca gtgaactctc ctgaatcttc gcctgcttct tcattctcct
     2941 tcgtttagct aatagaataa ctgctgagtt gtgaacagta aggtgtatgt gaggtgctcg
     3001 aaaacaaggt ttcaggtgac gcccccagaa taaaatttgg acggggggtt cagtggtggc
     3061 attgtgctat gacaccaata taaccctcac aaaccccttg ggcaataaat actagtgtag
     3121 gaatgaaaca ttctgaatat ctttaacaat agaaatccat ggggtgggga caagccgtaa
     3181 agactggatg tccatctcac acgaatttat ggctatgggc aacacataat cctagtgcaa
     3241 tatgatactg gggttattaa gatgtgtccc aggcagggac caagacaggt gaaccatgtt
     3301 gttacactct atttgtaaca aggggaaaga gagtggacgc cgacagcagc ggactccact
     3361 ggttgtctct aacacccccg aaaattaaac ggggctccac gccaatgggg cccataaaca
     3421 aagacaagtg gccactcttt tttttgaaat tgtggagtgg gggcacgcgt cagcccccac
     3481 acgccgccct gcggttttgg actgtaaaat aagggtgtaa taacttggct gattgtaacc
     3541 ccgctaacca ctgcggtcaa accacttgcc cacaaaacca ctaatggcac cccggggaat
     3601 acctgcataa gtaggtgggc gggccaagat aggggcgcga ttgctgcgat ctggaggaca
     3661 aattacacac acttgcgcct gagcgccaag cacagggttg ttggtcctca tattcacgag
     3721 gtcgctgaga gcacggtggg ctaatgttgc catgggtagc atatactacc caaatatctg
     3781 gatagcatat gctatcctaa tctatatctg ggtagcatag gctatcctaa tctatatctg
     3841 ggtagcatat gctatcctaa tctatatctg ggtagtatat gctatcctaa tttatatctg
     3901 ggtagcatag gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg
     3961 ggtagtatat gctatcctaa tctgtatccg ggtagcatat gctatcctaa tagagattag
     4021 ggtagtatat gctatcctaa tttatatctg ggtagcatat actacccaaa tatctggata
     4081 gcatatgcta tcctaatcta tatctgggta gcatatgcta tcctaatcta tatctgggta
     4141 gcataggcta tcctaatcta tatctgggta gcatatgcta tcctaatcta tatctgggta
     4201 gtatatgcta tcctaattta tatctgggta gcataggcta tcctaatcta tatctgggta
     4261 gcatatgcta tcctaatcta tatctgggta gtatatgcta tcctaatctg tatccgggta
     4321 gcatatgcta tcctcatgat aagctgtcaa acatgagaat tttcttgaag acgaaagggc
     4381 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     4441 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4501 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4561 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     4621 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     4681 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     4741 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     4801 gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     4861 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     4921 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     4981 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     5041 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     5101 accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     5161 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     5221 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     5281 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     5341 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     5401 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     5461 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5521 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5581 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5641 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5701 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     5761 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5821 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5881 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5941 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     6001 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6061 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6121 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6181 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6241 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     6301 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     6361 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     6421 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     6481 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     6541 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     6601 gccaagctct agctagaggt cgaccaattc tcatgtttga cagcttatca tcgcagatcc
     6661 gggcaacgtt gttgccattg ctgcaggcgc agaactggta ggtatggaag atctatacat
     6721 tgaatcaata ttggcaatta gccatattag tcattggtta tatagcataa atcaatattg
     6781 gctattggcc attgcatacg ttgtatctat atcataatat gtacatttat attggctcat
     6841 gtccaatatg accgc
//
