LOCUS       sequence-262378-        6552 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 6552)
  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 6552)
  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: D86_Ld_260650
FEATURES             Location/Qualifiers
     source          1..6552
                     /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             1716..1733
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1785..1840
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1839..1858)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4076..4094)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4162..4266
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4267..4335,4336..5127)
                     /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(4485..4504)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5298..5886
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5787..5806
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6040..6057
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6174..6195
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6210..6215,6216..6233,6234..6240)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6248..6264
                     /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     6253..6275
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6272..6288
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6272..6288
                     /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 aagaaggaac ggttacatac tacggtcagc tgagattgcc gccaacgtac ctgcgtcatc
     1261 cagattgctt tgaggccctc aacgatatcc agcccggcag cgtgctgttg tataacggcc
     1321 agcacagatt tgtcgtgcca actgccaggg acggctcctt ctccgtctat aagctgccct
     1381 atgggacata cattctgcag gccgagtatc attactttat gttccccacc gtgcgcgtcg
     1441 aggttatgta ccgcgatacc ggcgacgacc aaaaggagac ttttatacgt acatcagcta
     1501 acgactatcc cgtgcggcat ctcgaggggt ctggactgga cgaggaatca ccagccgtga
     1561 ttccctttag cggttaccat aactactata tcccccgcca acagatggac attgtttccg
     1621 gcgcgcccgg cagcctgcac cacatcctgg acgcccagaa gatgctgtgg aaccacaggg
     1681 acaggaacct gcctcctctg gcccctctgg gcccccacca ccatcaccac cattgagtct
     1741 agacacgtga ttaattaagg atcccccgac ctcgacctct ggctaataaa ggaaatttat
     1801 tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag
     1861 ggcaaatcat ttggtcgaga tccctcggag atctctagct agaggatcga tccccgcccc
     1921 ggacgaacta aacctgacta cgacatctct gccccttctt cgcggggcag tgcatgtaat
     1981 cccttcagtt ggttggtaca acttgccaac tgggccctgt tccacatgtg acacgggggg
     2041 ggaccaaaca caaaggggtt ctctgactgt agttgacatc cttataaatg gatgtgcaca
     2101 tttgccaaca ctgagtggct ttcatcctgg agcagacttt gcagtctgtg gactgcaaca
     2161 caacattgcc tttatgtgta actcttggct gaagctctta caccaatgct gggggacatg
     2221 tacctcccag gggcccagga agactacggg aggctacacc aacgtcaatc agaggggcct
     2281 gtgtagctac cgataagcgg accctcaaga gggcattagc aatagtgttt ataaggcccc
     2341 cttgttaacc ctaaacgggt agcatatgct tcccgggtag tagtatatac tatccagact
     2401 aaccctaatt caatagcata tgttacccaa cgggaagcat atgctatcga attagggtta
     2461 gtaaaagggt cctaaggaac agcgatatct cccaccccat gagctgtcac ggttttattt
     2521 acatggggtc aggattccac gagggtagtg aaccatttta gtcacaaggg cagtggctga
     2581 agatcaagga gcgggcagtg aactctcctg aatcttcgcc tgcttcttca ttctccttcg
     2641 tttagctaat agaataactg ctgagttgtg aacagtaagg tgtatgtgag gtgctcgaaa
     2701 acaaggtttc aggtgacgcc cccagaataa aatttggacg gggggttcag tggtggcatt
     2761 gtgctatgac accaatataa ccctcacaaa ccccttgggc aataaatact agtgtaggaa
     2821 tgaaacattc tgaatatctt taacaataga aatccatggg gtggggacaa gccgtaaaga
     2881 ctggatgtcc atctcacacg aatttatggc tatgggcaac acataatcct agtgcaatat
     2941 gatactgggg ttattaagat gtgtcccagg cagggaccaa gacaggtgaa ccatgttgtt
     3001 acactctatt tgtaacaagg ggaaagagag tggacgccga cagcagcgga ctccactggt
     3061 tgtctctaac acccccgaaa attaaacggg gctccacgcc aatggggccc ataaacaaag
     3121 acaagtggcc actctttttt ttgaaattgt ggagtggggg cacgcgtcag cccccacacg
     3181 ccgccctgcg gttttggact gtaaaataag ggtgtaataa cttggctgat tgtaaccccg
     3241 ctaaccactg cggtcaaacc acttgcccac aaaaccacta atggcacccc ggggaatacc
     3301 tgcataagta ggtgggcggg ccaagatagg ggcgcgattg ctgcgatctg gaggacaaat
     3361 tacacacact tgcgcctgag cgccaagcac agggttgttg gtcctcatat tcacgaggtc
     3421 gctgagagca cggtgggcta atgttgccat gggtagcata tactacccaa atatctggat
     3481 agcatatgct atcctaatct atatctgggt agcataggct atcctaatct atatctgggt
     3541 agcatatgct atcctaatct atatctgggt agtatatgct atcctaattt atatctgggt
     3601 agcataggct atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt
     3661 agtatatgct atcctaatct gtatccgggt agcatatgct atcctaatag agattagggt
     3721 agtatatgct atcctaattt atatctgggt agcatatact acccaaatat ctggatagca
     3781 tatgctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagca
     3841 taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagta
     3901 tatgctatcc taatttatat ctgggtagca taggctatcc taatctatat ctgggtagca
     3961 tatgctatcc taatctatat ctgggtagta tatgctatcc taatctgtat ccgggtagca
     4021 tatgctatcc tcatgataag ctgtcaaaca tgagaatttt cttgaagacg aaagggcctc
     4081 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     4141 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     4201 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     4261 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     4321 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     4381 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     4441 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     4501 ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     4561 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     4621 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     4681 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     4741 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     4801 acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     4861 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     4921 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     4981 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     5041 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     5101 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     5161 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     5221 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     5281 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     5341 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     5401 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     5461 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     5521 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     5581 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     5641 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     5701 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     5761 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     5821 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     5881 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     5941 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     6001 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     6061 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     6121 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     6181 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     6241 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     6301 aagctctagc tagaggtcga ccaattctca tgtttgacag cttatcatcg cagatccggg
     6361 caacgttgtt gccattgctg caggcgcaga actggtaggt atggaagatc tatacattga
     6421 atcaatattg gcaattagcc atattagtca ttggttatat agcataaatc aatattggct
     6481 attggccatt gcatacgttg tatctatatc ataatatgta catttatatt ggctcatgtc
     6541 caatatgacc gc
//
