LOCUS       sequence-262331-        7353 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 7353)
  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 7353)
  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: D38_Ld_341960
FEATURES             Location/Qualifiers
     source          1..7353
                     /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             2517..2534
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2586..2641
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2640..2659)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4877..4895)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4963..5067
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5068..5136,5137..5928)
                     /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(5286..5305)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6099..6687
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6588..6607
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6841..6858
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6975..6996
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7011..7016,7017..7034,7035..7041)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7049..7065
                     /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     7054..7076
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7073..7089
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7073..7089
                     /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 cctaccctta cggcagatct agcgccgtga ccgagctgac ccctgccagc ctgcacgcct
     1261 tcgtgaacac ccacaagccc gtggtcatcc tgttctacgc cccttggtgc ggccactgca
     1321 agcagttcca ccccgagtac gagagattcg ccgagagcgt gaagggcacc atcagagtgg
     1381 gcgccatcga cgccgacaag aatgccgaca tcggccagca gttcggcgtg cggggcttcc
     1441 ccaccatcaa gtactggaag tccggcacca agagcgtgtc cagcagccag gaataccagg
     1501 gccagagaac cgccgctgcc ctgcagtctc tggtggtgga aggcatcagc agcagcaaag
     1561 tgatgaccgt gaccaccgcc gagcagatga agcaggccgc cagggacgcc cccaagaaaa
     1621 tgatcggagt gctgctgagc gccaagaaca aggtgccccc catgttctcc gtgatgggca
     1681 tgagccccaa gctgaaagaa ctgcccctgg tgttcgccgg cagcagctcc cctgagaagg
     1741 gcgtggccaa ggccctggga gtgtcccagc tgcccatgat tggcgtgctg cggtacaccc
     1801 cagccgaggg ggacgccgaa gagaagttcg agatggtgcc ctatgccaag aaggccattg
     1861 cctacgagcc tgtggcccgg ttcttcctgg actgcgccga gaaagagtgc agcgccggca
     1921 ccgaggcccc catgaattcc gtgtccctgg acgaggacga gagccagcac cagacccggg
     1981 aagccgccag acagtctagc gtggccctgc ccgtggaacc cgtcgagtac acacctaaag
     2041 tgatcagcct gttctgtgcc cctcagacca gaaagatcag aggccagagc cccctgtgcg
     2101 tgttcagcct gagcgagcac gtgaacctgg acaacgtgca ccagtacttc aagaacgacc
     2161 ccctgctgtt cttcgaggcc tcccagcaca aagaggaact gctccagcag ctgcaggccg
     2221 actttggcgt gcagatcgcc agcaccggcg aggccatgac aagcctggcc ggctccatcg
     2281 tggtgctgag acacagcaag cccaccaccg tcagatacgc cctgctggaa aacatcgaca
     2341 gcgacgacga cctgcaggct gctctgcagc ggatcctgaa cggcgaggtg cacctgagat
     2401 ccgagagagt gggagccgct ggcgcgcccg gcagcctgca ccacatcctg gacgcccaga
     2461 agatgctgtg gaaccacagg gacaggaacc tgcctcctct ggcccctctg ggcccccacc
     2521 accatcacca ccattgagtc tagacacgtg attaattaag gatcccccga cctcgacctc
     2581 tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc
     2641 actcggaagg acatatggga gggcaaatca tttggtcgag atccctcgga gatctctagc
     2701 tagaggatcg atccccgccc cggacgaact aaacctgact acgacatctc tgccccttct
     2761 tcgcggggca gtgcatgtaa tcccttcagt tggttggtac aacttgccaa ctgggccctg
     2821 ttccacatgt gacacggggg gggaccaaac acaaaggggt tctctgactg tagttgacat
     2881 ccttataaat ggatgtgcac atttgccaac actgagtggc tttcatcctg gagcagactt
     2941 tgcagtctgt ggactgcaac acaacattgc ctttatgtgt aactcttggc tgaagctctt
     3001 acaccaatgc tgggggacat gtacctccca ggggcccagg aagactacgg gaggctacac
     3061 caacgtcaat cagaggggcc tgtgtagcta ccgataagcg gaccctcaag agggcattag
     3121 caatagtgtt tataaggccc ccttgttaac cctaaacggg tagcatatgc ttcccgggta
     3181 gtagtatata ctatccagac taaccctaat tcaatagcat atgttaccca acgggaagca
     3241 tatgctatcg aattagggtt agtaaaaggg tcctaaggaa cagcgatatc tcccacccca
     3301 tgagctgtca cggttttatt tacatggggt caggattcca cgagggtagt gaaccatttt
     3361 agtcacaagg gcagtggctg aagatcaagg agcgggcagt gaactctcct gaatcttcgc
     3421 ctgcttcttc attctccttc gtttagctaa tagaataact gctgagttgt gaacagtaag
     3481 gtgtatgtga ggtgctcgaa aacaaggttt caggtgacgc ccccagaata aaatttggac
     3541 ggggggttca gtggtggcat tgtgctatga caccaatata accctcacaa accccttggg
     3601 caataaatac tagtgtagga atgaaacatt ctgaatatct ttaacaatag aaatccatgg
     3661 ggtggggaca agccgtaaag actggatgtc catctcacac gaatttatgg ctatgggcaa
     3721 cacataatcc tagtgcaata tgatactggg gttattaaga tgtgtcccag gcagggacca
     3781 agacaggtga accatgttgt tacactctat ttgtaacaag gggaaagaga gtggacgccg
     3841 acagcagcgg actccactgg ttgtctctaa cacccccgaa aattaaacgg ggctccacgc
     3901 caatggggcc cataaacaaa gacaagtggc cactcttttt tttgaaattg tggagtgggg
     3961 gcacgcgtca gcccccacac gccgccctgc ggttttggac tgtaaaataa gggtgtaata
     4021 acttggctga ttgtaacccc gctaaccact gcggtcaaac cacttgccca caaaaccact
     4081 aatggcaccc cggggaatac ctgcataagt aggtgggcgg gccaagatag gggcgcgatt
     4141 gctgcgatct ggaggacaaa ttacacacac ttgcgcctga gcgccaagca cagggttgtt
     4201 ggtcctcata ttcacgaggt cgctgagagc acggtgggct aatgttgcca tgggtagcat
     4261 atactaccca aatatctgga tagcatatgc tatcctaatc tatatctggg tagcataggc
     4321 tatcctaatc tatatctggg tagcatatgc tatcctaatc tatatctggg tagtatatgc
     4381 tatcctaatt tatatctggg tagcataggc tatcctaatc tatatctggg tagcatatgc
     4441 tatcctaatc tatatctggg tagtatatgc tatcctaatc tgtatccggg tagcatatgc
     4501 tatcctaata gagattaggg tagtatatgc tatcctaatt tatatctggg tagcatatac
     4561 tacccaaata tctggatagc atatgctatc ctaatctata tctgggtagc atatgctatc
     4621 ctaatctata tctgggtagc ataggctatc ctaatctata tctgggtagc atatgctatc
     4681 ctaatctata tctgggtagt atatgctatc ctaatttata tctgggtagc ataggctatc
     4741 ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt atatgctatc
     4801 ctaatctgta tccgggtagc atatgctatc ctcatgataa gctgtcaaac atgagaattt
     4861 tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     4921 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     4981 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     5041 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     5101 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     5161 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     5221 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     5281 gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc
     5341 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     5401 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     5461 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     5521 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     5581 ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta
     5641 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     5701 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     5761 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     5821 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     5881 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     5941 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     6001 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     6061 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     6121 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     6181 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     6241 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     6301 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     6361 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     6421 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     6481 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     6541 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     6601 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     6661 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     6721 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     6781 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     6841 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     6901 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     6961 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
     7021 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     7081 agctatgacc atgattacgc caagctctag ctagaggtcg accaattctc atgtttgaca
     7141 gcttatcatc gcagatccgg gcaacgttgt tgccattgct gcaggcgcag aactggtagg
     7201 tatggaagat ctatacattg aatcaatatt ggcaattagc catattagtc attggttata
     7261 tagcataaat caatattggc tattggccat tgcatacgtt gtatctatat cataatatgt
     7321 acatttatat tggctcatgt ccaatatgac cgc
//
