LOCUS       sequence-262319-        6603 bp    DNA     circular SYN 15-APR-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 6603)
  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 6603)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: D24_Ld_171460
FEATURES             Location/Qualifiers
     source          1..6603
                     /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             1767..1784
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1836..1891
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1890..1909)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4127..4145)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4213..4317
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4318..4386,4387..5178)
                     /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(4536..4555)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5349..5937
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5838..5857
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6091..6108
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6225..6246
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6261..6266,6267..6284,6285..6291)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6299..6315
                     /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     6304..6326
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6323..6339
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6323..6339
                     /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 ccgtgatcag cagctgtgat gtgtgcggcc tgagcatcgt gaagcaggtc atggccccca
     1261 gcgacagatc cgatggcggc gtgccagagg ccacagccgt gcggtctagc cccagcaccg
     1321 ccacagagac acccatgctg gaactggacg accggcctag aggcgccgct gccggcaagg
     1381 ccaacaaata cgtggaagag ttctaccagg ccgctggcca cctgagatgc cctcagctgc
     1441 agcggtacac ccaggaagag gtcgcccggc accggaccaa ggacgacctg tggatcgtgg
     1501 tggacagcaa cgtgctgaac gtgtccgcct tcgtgcccca ccatcctggc ggcgacgtga
     1561 tcctggatgg cgtgggcggc caggacatgg ccacagtgtt cgcctacttc cacgacccct
     1621 ccaccgtgcg gctgatgctg agcttctgca tcggccggat cctgctgcag ggcgcgcccg
     1681 gcagcctgca ccacatcctg gacgcccaga agatgctgtg gaaccacagg gacaggaacc
     1741 tgcctcctct ggcccctctg ggcccccacc accatcacca ccattgagtc tagacacgtg
     1801 attaattaag gatcccccga cctcgacctc tggctaataa aggaaattta ttttcattgc
     1861 aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca
     1921 tttggtcgag atccctcgga gatctctagc tagaggatcg atccccgccc cggacgaact
     1981 aaacctgact acgacatctc tgccccttct tcgcggggca gtgcatgtaa tcccttcagt
     2041 tggttggtac aacttgccaa ctgggccctg ttccacatgt gacacggggg gggaccaaac
     2101 acaaaggggt tctctgactg tagttgacat ccttataaat ggatgtgcac atttgccaac
     2161 actgagtggc tttcatcctg gagcagactt tgcagtctgt ggactgcaac acaacattgc
     2221 ctttatgtgt aactcttggc tgaagctctt acaccaatgc tgggggacat gtacctccca
     2281 ggggcccagg aagactacgg gaggctacac caacgtcaat cagaggggcc tgtgtagcta
     2341 ccgataagcg gaccctcaag agggcattag caatagtgtt tataaggccc ccttgttaac
     2401 cctaaacggg tagcatatgc ttcccgggta gtagtatata ctatccagac taaccctaat
     2461 tcaatagcat atgttaccca acgggaagca tatgctatcg aattagggtt agtaaaaggg
     2521 tcctaaggaa cagcgatatc tcccacccca tgagctgtca cggttttatt tacatggggt
     2581 caggattcca cgagggtagt gaaccatttt agtcacaagg gcagtggctg aagatcaagg
     2641 agcgggcagt gaactctcct gaatcttcgc ctgcttcttc attctccttc gtttagctaa
     2701 tagaataact gctgagttgt gaacagtaag gtgtatgtga ggtgctcgaa aacaaggttt
     2761 caggtgacgc ccccagaata aaatttggac ggggggttca gtggtggcat tgtgctatga
     2821 caccaatata accctcacaa accccttggg caataaatac tagtgtagga atgaaacatt
     2881 ctgaatatct ttaacaatag aaatccatgg ggtggggaca agccgtaaag actggatgtc
     2941 catctcacac gaatttatgg ctatgggcaa cacataatcc tagtgcaata tgatactggg
     3001 gttattaaga tgtgtcccag gcagggacca agacaggtga accatgttgt tacactctat
     3061 ttgtaacaag gggaaagaga gtggacgccg acagcagcgg actccactgg ttgtctctaa
     3121 cacccccgaa aattaaacgg ggctccacgc caatggggcc cataaacaaa gacaagtggc
     3181 cactcttttt tttgaaattg tggagtgggg gcacgcgtca gcccccacac gccgccctgc
     3241 ggttttggac tgtaaaataa gggtgtaata acttggctga ttgtaacccc gctaaccact
     3301 gcggtcaaac cacttgccca caaaaccact aatggcaccc cggggaatac ctgcataagt
     3361 aggtgggcgg gccaagatag gggcgcgatt gctgcgatct ggaggacaaa ttacacacac
     3421 ttgcgcctga gcgccaagca cagggttgtt ggtcctcata ttcacgaggt cgctgagagc
     3481 acggtgggct aatgttgcca tgggtagcat atactaccca aatatctgga tagcatatgc
     3541 tatcctaatc tatatctggg tagcataggc tatcctaatc tatatctggg tagcatatgc
     3601 tatcctaatc tatatctggg tagtatatgc tatcctaatt tatatctggg tagcataggc
     3661 tatcctaatc tatatctggg tagcatatgc tatcctaatc tatatctggg tagtatatgc
     3721 tatcctaatc tgtatccggg tagcatatgc tatcctaata gagattaggg tagtatatgc
     3781 tatcctaatt tatatctggg tagcatatac tacccaaata tctggatagc atatgctatc
     3841 ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagc ataggctatc
     3901 ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt atatgctatc
     3961 ctaatttata tctgggtagc ataggctatc ctaatctata tctgggtagc atatgctatc
     4021 ctaatctata tctgggtagt atatgctatc ctaatctgta tccgggtagc atatgctatc
     4081 ctcatgataa gctgtcaaac atgagaattt tcttgaagac gaaagggcct cgtgatacgc
     4141 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     4201 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4261 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     4321 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     4381 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     4441 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     4501 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     4561 gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     4621 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     4681 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     4741 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     4801 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     4861 gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     4921 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     4981 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     5041 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     5101 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     5161 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     5221 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5281 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5341 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5401 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5461 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     5521 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     5581 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     5641 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     5701 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     5761 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     5821 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     5881 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     5941 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     6001 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     6061 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     6121 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     6181 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     6241 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     6301 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagctctag
     6361 ctagaggtcg accaattctc atgtttgaca gcttatcatc gcagatccgg gcaacgttgt
     6421 tgccattgct gcaggcgcag aactggtagg tatggaagat ctatacattg aatcaatatt
     6481 ggcaattagc catattagtc attggttata tagcataaat caatattggc tattggccat
     6541 tgcatacgtt gtatctatat cataatatgt acatttatat tggctcatgt ccaatatgac
     6601 cgc
//
