LOCUS       sequence-262320-       10119 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 10119)
  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 10119)
  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: D25_Ld_190160
FEATURES             Location/Qualifiers
     source          1..10119
                     /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             5283..5300
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    5352..5407
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5406..5425)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(7643..7661)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7729..7833
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7834..7902,7903..8694)
                     /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(8052..8071)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8865..9453
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9354..9373
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9607..9624
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9741..9762
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9777..9782,9783..9800,9801..9807)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9815..9831
                     /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     9820..9842
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9839..9855
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9839..9855
                     /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 ccgctctgac agcccctttt ggcggctgtg aagccggcac cagcggctac gccattcaca
     1261 gccctggcga gaacgacaag ggcgcccagt acctgatgga caacggaatc gccttcgcca
     1321 tcctgcctgc caccgtgacc aacggcgtgg tcacctggga gaatggcgag gtggccgcct
     1381 ggatgccccc cgattttcag gcccctagcg acgcccacta ctgcctgatc ctgcggaaga
     1441 acggcgcttg gagcgtggtg tccatgccct tcacctggaa ccacctgtgc atcgacggca
     1501 gagaggacgg cgagacaacc accaccgcca cccagatgcc caacaccacc acaacctcta
     1561 ccaccactac cacccctgcc cctaccgcca acccccacct gagcagctcc agcttccaga
     1621 gcgagggcag cgccagcagc agcttcgcca gccccatcac aacaaccaca ctggccccca
     1681 gccctaccac ctccacaaca gccgcccctc cacctagcgt gcccaccgtg tctagcgtga
     1741 tgcagaccgt gcgctccgtg gtgttcgccg ccgtgcctac agtgaacgtg atggacttca
     1801 ccagcaacgt ggtgtacctg ccagccacag ccaccctggt ggaatccatg caggccctgc
     1861 aggccgccag catcgtgatc gccgtcagat ttggcgccac cggccctgtg tccgccctgc
     1921 tgacagatgc caccgccgcc tccgtgtcct tccacgtgcc ccacgagctg tacaacgtgt
     1981 ccgtcgtgac cagccacgcc ctgagcctgg tcaccctgaa caccgccgat gtgctggtcg
     2041 tgggcagcca gctgagaatc acccccgtgc tgatcgctct gcagcagatg agcatgggcg
     2101 tggtgctgag cgacgagagc atcctggaag gcaccgacat cctggccaca gtccgctttg
     2161 ccggcgctgg caccgcactg ctgtacacca ccgacgtgtc caaagtgatc acaagcaccc
     2221 tgagcttcag catggtcaac aacggcgccg tgggctcccg gtgcaagccc agcacaagca
     2281 gcatccctta cttcatcacc agatacgacg tgcggttcct ggtgcccggc aactacaccg
     2341 ccagagtgca cgccggcaga aagtccgtgt gcgccaccta caacggcaag tctgtggtgg
     2401 tggcccccct ggccatccag ggcattgccg ccacaaccac tgccgctagc tttcccgccg
     2461 ctacagccat cccaatccgg ctgcagggcg gcacaactac cggaattgcc agaggcgcct
     2521 acgccgcctt tgccagcgcc tccagcgatt gcacctccga gggcgccaag atcagcctgc
     2581 tgcccgtgaa cagcttcaac agctcctccg acagctctgc cgtgccaggc gctctgctgg
     2641 tggccagctc tgaggtgcca accggcagct acgtgtgcat gcggcggacc agcaccaata
     2701 ccaagttcgc cgtgcggtat cacggcaagc ccgtgaccgt gggagccgcc tctatgcctc
     2761 ctgccgctgc tgccactgcc cctgtgctga ccatcttcaa cggcagctgc ggctccctga
     2821 acctgctgga atttgtgcgg cctcccgcca gcgaggacga tatcgccgcc tacctgagcg
     2881 ccgcttccaa tgtgtccgag gcctaccggg tgttcagatt cgtggccgtg aaagccggcg
     2941 caactgccgc cgataaggcc gacgcctgcc ggcggtttaa gcaggaagcc accagggacg
     3001 ccgccaatgg cgccatgcag ttcagcttcc cattctatct gcagcagacc aatggcgtgc
     3061 tgtgcgccga cgaggaatac gtgcggatcg attacaccgt gatccagccc accaactggc
     3121 tgatgtccat ggccggcgtg tccgacttcg cctcccggat gctggtgctg tccagcggcc
     3181 ctgagtacga cgctgtgcct ctgagcgtgc ggagcagcag gctggcctct gtgggctctg
     3241 gcacagccgt caggatcagc caggacggct cctgcgcctt cggctacaca gcccggatcc
     3301 agagctacaa cacccagcag tacattgccg tccccactgc cagcgtgggc gtgtacacac
     3361 tgtgtgccgg cctgtacaat gagacaggca cctccatgtt cgtgtccacc ggctcccagg
     3421 tggacctggc ctcctatgtg cggggcagac ggaagtggat gcacgaggcc ggcatcgaga
     3481 gaaccgagtg gaccccctgt ggctccgcct ccacatgtgg cctgcacaca cagcaggcct
     3541 acatcgaagt gtgcaacggg gctgtgcgga gaaacgtgtg gccccagaga ctggacggcg
     3601 cctttgtggg aagcgacagc gccatggtcc tgatggccgg aaatctgtac aagacccagg
     3661 ccattgtgct gatgaacgac gtgtacgtgc ccgtggccgc tgccttcgct cactgtcagc
     3721 agttctccct gctgcagctg agcaccgacg acggcgctac ctatggccca gccgccgtcg
     3781 tgctgggcaa ccaccctcag atcgccctgt ccccagtgac tgatgtctcc ggcgtgatcg
     3841 tgggcttcgt gccctggaac gccatgtgca gaggcgtgca cacaggcgct gatgtgatgg
     3901 tggctcggac cgccgctggc atctccgtga tcgacatcag cggcctgctg tttaccaagt
     3961 ctgagggcta ctacgccgtg tgtgtgcgga acggggtggc ctgggctaga gtgaccgaca
     4021 cctacatcaa ggtggtcatc ggcggcgtga cagtgtacca ggccgaggcc atcaacaccg
     4081 gcgacgctct ggccaaccag aagtccatcc acctgtacca gacacagacc gccacatggg
     4141 gcgtggccgg cacattcagc ccttccgtgt ctctgctgct gaagctcgtg tggatggaca
     4201 gcacatgcac catggatgcc gcctataccg ctgccctgcg gtacatcagc cctagcagcg
     4261 cccatgccac agtggaagcc agcctgattg ctgccgcccc agctggcgtg gcatccccca
     4321 ccctgtaccc ctgctacatg gccaacacct tcggcaatac cgccgcaccc atgcctttcg
     4381 aggaacacag aggccggtcc atcacagtga cccagctgac cctgtcctcc ctgttctacc
     4441 tgccctatct ggaactgtct gtgtctgccc ccgtggacac ccggattctg ctgaccgagc
     4501 cccaggaccc ccctatcacc cgggtgtccc tgagagtgac aagctcccag tctgatgtgc
     4561 cccagtgtgg cgccgcagcc acatacgcta cagtgctgac aatcgagact gacgacctgg
     4621 gcaggcggtg gatgaccgtg cctgcctggg tgccacagac actgggcggc agcggcgcca
     4681 caaatgtgta tctgaagatg tgcgcaaccg cctcctctag cgccaatccc gagtttgtgg
     4741 ccgtcgaagc ctacctcgtg ctgtctgcca acgccaccaa cctcgccggc aaccccatga
     4801 gcctgcggct gaccttcctg cctctcaaca gcagcaccga tctgctggac tacaaggccg
     4861 ctcagtatgc cctcgtgaag tacgccgcaa ccgagctgaa ggtgcccgag acagtggtgc
     4921 tcgtggctag cctgggaaac aacgccttcg agctgttcat cgatgacgcc ctccagaccg
     4981 gcgcagcccc tcagagcaca ctgaccccta ctcagcagct gtatcagacc ctggctaagg
     5041 ggaccaatct gcctctgttc ctgaacggcc gggataacgc cacagcttgg tttgccctgg
     5101 caaaggctga ggccgtgcac ctcgtggacc agacctccgt ggacgattac cccgccctgg
     5161 ccctgccagt gtctggatct gccggaggcg cgcccggcag cctgcaccac atcctggacg
     5221 cccagaagat gctgtggaac cacagggaca ggaacctgcc tcctctggcc cctctgggcc
     5281 cccaccacca tcaccaccat tgagtctaga cacgtgatta attaaggatc ccccgacctc
     5341 gacctctggc taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg
     5401 tctctcactc ggaaggacat atgggagggc aaatcatttg gtcgagatcc ctcggagatc
     5461 tctagctaga ggatcgatcc ccgccccgga cgaactaaac ctgactacga catctctgcc
     5521 ccttcttcgc ggggcagtgc atgtaatccc ttcagttggt tggtacaact tgccaactgg
     5581 gccctgttcc acatgtgaca cgggggggga ccaaacacaa aggggttctc tgactgtagt
     5641 tgacatcctt ataaatggat gtgcacattt gccaacactg agtggctttc atcctggagc
     5701 agactttgca gtctgtggac tgcaacacaa cattgccttt atgtgtaact cttggctgaa
     5761 gctcttacac caatgctggg ggacatgtac ctcccagggg cccaggaaga ctacgggagg
     5821 ctacaccaac gtcaatcaga ggggcctgtg tagctaccga taagcggacc ctcaagaggg
     5881 cattagcaat agtgtttata aggccccctt gttaacccta aacgggtagc atatgcttcc
     5941 cgggtagtag tatatactat ccagactaac cctaattcaa tagcatatgt tacccaacgg
     6001 gaagcatatg ctatcgaatt agggttagta aaagggtcct aaggaacagc gatatctccc
     6061 accccatgag ctgtcacggt tttatttaca tggggtcagg attccacgag ggtagtgaac
     6121 cattttagtc acaagggcag tggctgaaga tcaaggagcg ggcagtgaac tctcctgaat
     6181 cttcgcctgc ttcttcattc tccttcgttt agctaataga ataactgctg agttgtgaac
     6241 agtaaggtgt atgtgaggtg ctcgaaaaca aggtttcagg tgacgccccc agaataaaat
     6301 ttggacgggg ggttcagtgg tggcattgtg ctatgacacc aatataaccc tcacaaaccc
     6361 cttgggcaat aaatactagt gtaggaatga aacattctga atatctttaa caatagaaat
     6421 ccatggggtg gggacaagcc gtaaagactg gatgtccatc tcacacgaat ttatggctat
     6481 gggcaacaca taatcctagt gcaatatgat actggggtta ttaagatgtg tcccaggcag
     6541 ggaccaagac aggtgaacca tgttgttaca ctctatttgt aacaagggga aagagagtgg
     6601 acgccgacag cagcggactc cactggttgt ctctaacacc cccgaaaatt aaacggggct
     6661 ccacgccaat ggggcccata aacaaagaca agtggccact cttttttttg aaattgtgga
     6721 gtgggggcac gcgtcagccc ccacacgccg ccctgcggtt ttggactgta aaataagggt
     6781 gtaataactt ggctgattgt aaccccgcta accactgcgg tcaaaccact tgcccacaaa
     6841 accactaatg gcaccccggg gaatacctgc ataagtaggt gggcgggcca agataggggc
     6901 gcgattgctg cgatctggag gacaaattac acacacttgc gcctgagcgc caagcacagg
     6961 gttgttggtc ctcatattca cgaggtcgct gagagcacgg tgggctaatg ttgccatggg
     7021 tagcatatac tacccaaata tctggatagc atatgctatc ctaatctata tctgggtagc
     7081 ataggctatc ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt
     7141 atatgctatc ctaatttata tctgggtagc ataggctatc ctaatctata tctgggtagc
     7201 atatgctatc ctaatctata tctgggtagt atatgctatc ctaatctgta tccgggtagc
     7261 atatgctatc ctaatagaga ttagggtagt atatgctatc ctaatttata tctgggtagc
     7321 atatactacc caaatatctg gatagcatat gctatcctaa tctatatctg ggtagcatat
     7381 gctatcctaa tctatatctg ggtagcatag gctatcctaa tctatatctg ggtagcatat
     7441 gctatcctaa tctatatctg ggtagtatat gctatcctaa tttatatctg ggtagcatag
     7501 gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagtatat
     7561 gctatcctaa tctgtatccg ggtagcatat gctatcctca tgataagctg tcaaacatga
     7621 gaattttctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     7681 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     7741 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     7801 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     7861 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     7921 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     7981 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     8041 tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc
     8101 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     8161 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     8221 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     8281 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     8341 gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac aacgttgcgc
     8401 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     8461 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     8521 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     8581 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     8641 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     8701 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     8761 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     8821 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     8881 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     8941 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     9001 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     9061 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     9121 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     9181 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     9241 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     9301 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     9361 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     9421 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     9481 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     9541 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     9601 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     9661 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     9721 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     9781 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     9841 ggaaacagct atgaccatga ttacgccaag ctctagctag aggtcgacca attctcatgt
     9901 ttgacagctt atcatcgcag atccgggcaa cgttgttgcc attgctgcag gcgcagaact
     9961 ggtaggtatg gaagatctat acattgaatc aatattggca attagccata ttagtcattg
    10021 gttatatagc ataaatcaat attggctatt ggccattgca tacgttgtat ctatatcata
    10081 atatgtacat ttatattggc tcatgtccaa tatgaccgc
//
