LOCUS       sequence-262323-        7299 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 7299)
  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 7299)
  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: D28_Ld_241170
FEATURES             Location/Qualifiers
     source          1..7299
                     /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             2463..2480
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2532..2587
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2586..2605)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4823..4841)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4909..5013
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5014..5082,5083..5874)
                     /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(5232..5251)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6045..6633
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6534..6553
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6787..6804
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6921..6942
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6957..6962,6963..6980,6981..6987)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6995..7011
                     /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     7000..7022
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7019..7035
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7019..7035
                     /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 ccgccagcat caccgtgacc gtgctgatca ccgagtgcct gttcagcagc cggcctcccc
     1261 ctagcccccc acctcatctg aaaggcgcct ggcagctgat gaaggccaca gtgacccggc
     1321 agctgtggtc cagaggcctg agactgccca gagtgtccag cgccgtgccc gccaacctga
     1381 gccactggga tggcctgcct cggaccatga caggcgccgc tgcttctgcc gctcctgccg
     1441 ccagagccag aaccacagcc agcctgacca gccccttcat ctgcctgcgg cggttcttct
     1501 ggaacccctt cgccagcggc accagaggca gcgtgtccgc caccagagac tccgtgcagc
     1561 ggctgaacgc cgagcaggac gcctctatcg ccgagaaggc ccaccgggac gccgacctgg
     1621 aaaacgagca gctgctgggc gcctgcctgg atctgctgga catgtgctcc ggcaacgccg
     1681 ctggcagcgc ccctagctct gccctggatg agctggcccg ggaccggatc tgggagctgg
     1741 tggacgtgct gctgcacgat caccacagag cccccgctga gcagttcgac ctggtgtaca
     1801 ccgccctgtg catgccagcc gcccaggacc acagacaggt gcagagaagc ctgctgctgg
     1861 tgctgcggag cgtggtgccc gagacactgt accgggtgtt cgagagcgtg gacctgttcg
     1921 tcgtgcggga cgacgagcag tccctgcacc agcgcgacgc cctcatgaag ttcgcccacg
     1981 cccagctggg agatctgcac gtgcccgacg gactggccga ggaagaggtg ctgagcgtct
     2041 acgtggacga catcaagacc gccttccccg tgctggccac ctgtcccgct tggcaggtcg
     2101 tggaaaggga cgccaccaca atcgccctga agtccaagct gttcgccctg ctggggcggc
     2161 tgtgcgccga gttcgatcag gaaaagaccg gcaaagtcaa gctggccgat ctgagaagca
     2221 ccgccgagcg ggtgctgggc accgatcagg cctctagact gctggaaggc gcccaggtgg
     2281 acaaggacgg caagatcgcc tatgcccagc tcgtggctct gctgtcccgg ccacctccca
     2341 agaagagggc ccctgccgaa gtgacaggcg cgcccggcag cctgcaccac atcctggacg
     2401 cccagaagat gctgtggaac cacagggaca ggaacctgcc tcctctggcc cctctgggcc
     2461 cccaccacca tcaccaccat tgagtctaga cacgtgatta attaaggatc ccccgacctc
     2521 gacctctggc taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg
     2581 tctctcactc ggaaggacat atgggagggc aaatcatttg gtcgagatcc ctcggagatc
     2641 tctagctaga ggatcgatcc ccgccccgga cgaactaaac ctgactacga catctctgcc
     2701 ccttcttcgc ggggcagtgc atgtaatccc ttcagttggt tggtacaact tgccaactgg
     2761 gccctgttcc acatgtgaca cgggggggga ccaaacacaa aggggttctc tgactgtagt
     2821 tgacatcctt ataaatggat gtgcacattt gccaacactg agtggctttc atcctggagc
     2881 agactttgca gtctgtggac tgcaacacaa cattgccttt atgtgtaact cttggctgaa
     2941 gctcttacac caatgctggg ggacatgtac ctcccagggg cccaggaaga ctacgggagg
     3001 ctacaccaac gtcaatcaga ggggcctgtg tagctaccga taagcggacc ctcaagaggg
     3061 cattagcaat agtgtttata aggccccctt gttaacccta aacgggtagc atatgcttcc
     3121 cgggtagtag tatatactat ccagactaac cctaattcaa tagcatatgt tacccaacgg
     3181 gaagcatatg ctatcgaatt agggttagta aaagggtcct aaggaacagc gatatctccc
     3241 accccatgag ctgtcacggt tttatttaca tggggtcagg attccacgag ggtagtgaac
     3301 cattttagtc acaagggcag tggctgaaga tcaaggagcg ggcagtgaac tctcctgaat
     3361 cttcgcctgc ttcttcattc tccttcgttt agctaataga ataactgctg agttgtgaac
     3421 agtaaggtgt atgtgaggtg ctcgaaaaca aggtttcagg tgacgccccc agaataaaat
     3481 ttggacgggg ggttcagtgg tggcattgtg ctatgacacc aatataaccc tcacaaaccc
     3541 cttgggcaat aaatactagt gtaggaatga aacattctga atatctttaa caatagaaat
     3601 ccatggggtg gggacaagcc gtaaagactg gatgtccatc tcacacgaat ttatggctat
     3661 gggcaacaca taatcctagt gcaatatgat actggggtta ttaagatgtg tcccaggcag
     3721 ggaccaagac aggtgaacca tgttgttaca ctctatttgt aacaagggga aagagagtgg
     3781 acgccgacag cagcggactc cactggttgt ctctaacacc cccgaaaatt aaacggggct
     3841 ccacgccaat ggggcccata aacaaagaca agtggccact cttttttttg aaattgtgga
     3901 gtgggggcac gcgtcagccc ccacacgccg ccctgcggtt ttggactgta aaataagggt
     3961 gtaataactt ggctgattgt aaccccgcta accactgcgg tcaaaccact tgcccacaaa
     4021 accactaatg gcaccccggg gaatacctgc ataagtaggt gggcgggcca agataggggc
     4081 gcgattgctg cgatctggag gacaaattac acacacttgc gcctgagcgc caagcacagg
     4141 gttgttggtc ctcatattca cgaggtcgct gagagcacgg tgggctaatg ttgccatggg
     4201 tagcatatac tacccaaata tctggatagc atatgctatc ctaatctata tctgggtagc
     4261 ataggctatc ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt
     4321 atatgctatc ctaatttata tctgggtagc ataggctatc ctaatctata tctgggtagc
     4381 atatgctatc ctaatctata tctgggtagt atatgctatc ctaatctgta tccgggtagc
     4441 atatgctatc ctaatagaga ttagggtagt atatgctatc ctaatttata tctgggtagc
     4501 atatactacc caaatatctg gatagcatat gctatcctaa tctatatctg ggtagcatat
     4561 gctatcctaa tctatatctg ggtagcatag gctatcctaa tctatatctg ggtagcatat
     4621 gctatcctaa tctatatctg ggtagtatat gctatcctaa tttatatctg ggtagcatag
     4681 gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagtatat
     4741 gctatcctaa tctgtatccg ggtagcatat gctatcctca tgataagctg tcaaacatga
     4801 gaattttctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     4861 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     4921 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4981 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     5041 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     5101 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     5161 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     5221 tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc
     5281 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     5341 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     5401 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     5461 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     5521 gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac aacgttgcgc
     5581 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     5641 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     5701 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     5761 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     5821 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     5881 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5941 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     6001 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     6061 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     6121 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     6181 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     6241 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     6301 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     6361 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6421 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6481 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6541 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6601 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6661 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     6721 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     6781 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     6841 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     6901 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     6961 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     7021 ggaaacagct atgaccatga ttacgccaag ctctagctag aggtcgacca attctcatgt
     7081 ttgacagctt atcatcgcag atccgggcaa cgttgttgcc attgctgcag gcgcagaact
     7141 ggtaggtatg gaagatctat acattgaatc aatattggca attagccata ttagtcattg
     7201 gttatatagc ataaatcaat attggctatt ggccattgca tacgttgtat ctatatcata
     7261 atatgtacat ttatattggc tcatgtccaa tatgaccgc
//
