LOCUS       sequence-274756-        6828 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 6828)
  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 6828)
  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: D77_Ld_352840
FEATURES             Location/Qualifiers
     source          1..6828
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     201..225
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     706..725
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      748..757
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1606..1623
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1675..1730
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1729..1748)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3966..3984)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4052..4156
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4157..4225,4226..5017)
                     /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(4375..4394)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5188..5776
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5677..5696
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5930..5947
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6064..6085
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6100..6105,6106..6123,6124..6130)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6138..6154
                     /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     6143..6165
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6162..6178
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6162..6178
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        6449..6828
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta caccaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcctcac tctcttccgc
      241 atcgctgtct gcgagggcca gctgttgggc tcgcggttga ggacaaactc ttcgcggtct
      301 ttccagtact cttggatcgg aaacccgtcg gcctccgaac ggtactccgc caccgaggga
      361 cctgagcgag tccgcatcga ccggatcgga aaacctctcg agaaaggcgt ctaaccagtc
      421 acagtcgcaa ggtaggctga gcaccgtggc gggcggcagc gggtggcggt cggggttgtt
      481 tctggcggag gtgctgctga tgatgtaatt aaagtaggcg gtcttgagac ggcggatggt
      541 cgaggtgagg tgtggcaggc ttgagatcca gctgttgggg tgagtactcc ctctcaaaag
      601 cgggcattac ttctgcgcta agattgtcag tttccaaaaa cgaggaggat ttgatattca
      661 cctggcccga tctggccata cacttgagtg acaatgacat ccactttgcc tttctctcca
      721 caggtgtcca ctcccaggtc caagtttgcc accatggagt ttcagaccca ggtactcatg
      781 tccctgctgc tctgcatgtc tggtgcggcc gcccaggtga ccgcaccttg gtatcgtatc
      841 aactataaca cccccgcata tgtcctgtat aagccccaag acgtcgccca gtatgcgaca
      901 agtagggcag ccgccaacgg gtactgtgtc cagaatggaa tgtccatctt ttctgcagaa
      961 ctccccgcca aggataacaa cttcgcttcc gacgcagcca aagaactggg cagcacagga
     1021 tactttacat acactggaag cagcgctgaa cccgtaccca gctttgctcg cttctgtaca
     1081 cagctgcaac ccaaccctag cagtactccc aattctatca aatgcgggtt tacttttaga
     1141 ttcggcctgc tcaccgtaat cgatactgtg cttctcggaa accagcccgg ggtggctcag
     1201 tattactcac tgtaccccaa tgtttggggc gccggcaatg ccaatacagg ctatgctgta
     1261 gagtgggata ccacaagacc caacccacgc gggtactatt tcatggccgt gaccggactc
     1321 actgacaccg tcgggcggca tgtgaacgta gatgcgaccg tttcagccag cgactcctat
     1381 gtgactgcct ctaaattcgc gatttactgt gaaagccaga gctttcttgg gtacgtgccc
     1441 gtgccgactg tgctggtgca gccaacattc gtgaaaggtt acaaggagct ggcctgggcc
     1501 caggaacatg gcgcgcccgg cagcctgcac cacatcctgg acgcccagaa gatgctgtgg
     1561 aaccacaggg acaggaacct gcctcctctg gcccctctgg gcccccacca ccatcaccac
     1621 cattgagtct agacacgtga ttaattaagg atcccccgac ctcgacctct ggctaataaa
     1681 ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga
     1741 catatgggag ggcaaatcat ttggtcgaga tccctcggag atctctagct agaggatcga
     1801 tccccgcccc ggacgaacta aacctgacta cgacatctct gccccttctt cgcggggcag
     1861 tgcatgtaat cccttcagtt ggttggtaca acttgccaac tgggccctgt tccacatgtg
     1921 acacgggggg ggaccaaaca caaaggggtt ctctgactgt agttgacatc cttataaatg
     1981 gatgtgcaca tttgccaaca ctgagtggct ttcatcctgg agcagacttt gcagtctgtg
     2041 gactgcaaca caacattgcc tttatgtgta actcttggct gaagctctta caccaatgct
     2101 gggggacatg tacctcccag gggcccagga agactacggg aggctacacc aacgtcaatc
     2161 agaggggcct gtgtggctac cgataagcgg accctcaaga gggcattagc aatagtgttt
     2221 ataaggcccc cttgttaacc ctaaacgggt agcatatgct tcccgggtag tagtatatac
     2281 tatccagact aaccctaatt caatagcata tgttacccaa cgggaagcat atgctatcga
     2341 attagggtta gtaaaagggt cctaaggaac agcgatatct cccaccccat gagctgtcac
     2401 ggttttattt acatggggtc aggattccac gagggtagtg aaccatttta gtcacaaggg
     2461 cagtggctga agatcaagga gcgggcagtg aactctcctg aatcttcgcc tgcttcttca
     2521 ttctccttcg tttagctaat agaataactg ctgagttgtg aacagtaagg tgtatgtgag
     2581 gtgctcgaaa acaaggtttc aggtgacgcc cccagaataa aatttggacg gggggttcag
     2641 tggtggcatt gtgctatgac accaatataa ccctcacaaa ccccttgggc aataaatact
     2701 agtgtaggaa tgaaacattc tgaatatctt taacaataga aatccatggg gtggggacaa
     2761 gccgtaaaga ctggatgtcc atctcacacg aatttatggc tatgggcaac acataatcct
     2821 agtgcaatat gatactgggg ttattaagat gtgtcccagg cagggaccaa gacaggtgaa
     2881 ccatgttgtt acactctatt tgtaacaagg ggaaagagag tggacgccga cagcagcgga
     2941 ctccactggt tgtctctaac acccccgaaa attaaacggg gctccacgcc aatggggccc
     3001 ataaacaaag acaagtggcc actctttttt ttgaaattgt ggagtggggg cacgcgtcag
     3061 cccccacacg ccgccctgcg gttttggact gtaaaataag ggtgtaataa cttggctgat
     3121 tgtaaccccg ctaaccactg cggtcaaacc acttgcccac aaaaccacta atggcacccc
     3181 ggggaatacc tgcataagta ggtgggcggg ccaagatagg ggcgcgattg ctgcgatctg
     3241 gaggacaaat tacacacact tgcgcctgag cgccaagcac agggttgttg gtcctcatat
     3301 tcacgaggtc gctgagagca cggtgggcta atgttgccat gggtagcata tactacccaa
     3361 atatctggat agcatatgct atcctaatct atatctgggt agcataggct atcctaatct
     3421 atatctgggt agcatatgct atcctaatct atatctgggt agtatatgct atcctaattt
     3481 atatctgggt agcataggct atcctaatct atatctgggt agcatatgct atcctaatct
     3541 atatctgggt agtatatgct atcctaatct gtatccgggt agcatatgct atcctaatag
     3601 agattagggt agtatatgct atcctaattt atatctgggt agcatatact acccaaatat
     3661 ctggatagca tatgctatcc taatctatat ctgggtagca tatgctatcc taatctatat
     3721 ctgggtagca taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat
     3781 ctgggtagta tatgctatcc taatttatat ctgggtagca taggctatcc taatctatat
     3841 ctgggtagca tatgctatcc taatctatat ctgggtagta tatgctatcc taatctgtat
     3901 ccgggtagca tatgctatcc tcatgataag ctgtcaaaca tgagaatttt cttgaagacg
     3961 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     4021 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4081 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4141 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     4201 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     4261 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     4321 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     4381 tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     4441 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     4501 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     4561 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     4621 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     4681 gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga
     4741 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     4801 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     4861 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     4921 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     4981 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     5041 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5101 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5161 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5221 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5281 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     5341 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5401 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5461 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     5521 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     5581 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     5641 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     5701 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     5761 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     5821 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     5881 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     5941 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     6001 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     6061 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     6121 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
     6181 tgattacgcc aagctctagc tagaggtcga ccaattctca tgtttgacag cttatcatcg
     6241 cagatccggg caacgttgtt gccattgctg caggcgcaga actggtaggt atggaagatc
     6301 tatacattga atcaatattg gcaattagcc atattagtca ttggttatat agcataaatc
     6361 aatattggct attggccatt gcatacgttg tatctatatc ataatatgta catttatatt
     6421 ggctcatgtc caatatgacc gccatgttga cattgattat tgactagtta ttaatagtaa
     6481 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     6541 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     6601 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     6661 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtcc gccccctatt
     6721 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttacgggac
     6781 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatg
//
