LOCUS       sequence-274856-        7248 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 7248)
  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 7248)
  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: D73_Ld_262620
FEATURES             Location/Qualifiers
     source          1..7248
                     /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             2026..2043
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2095..2150
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2149..2168)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4386..4404)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4472..4576
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4577..4645,4646..5437)
                     /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(4795..4814)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5608..6196
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6097..6116
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6350..6367
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6484..6505
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6520..6525,6526..6543,6544..6550)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6558..6574
                     /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     6563..6585
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6582..6598
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6582..6598
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        6869..7248
                     /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 gccgtgtcca ttccaagctt taatgaatcc
      841 cttctgggcg taaacgcatt taaccttaca tgttttgacg tgtacatgag aggctttggc
      901 acgagttgga tgcccaccag atcttatgcc gccatcactg acggcggcaa tattaggtgc
      961 agaacaaacc tgactgcatt cgcctactct ggggtggtgt gggcggacgt agtggtaaat
     1021 cctggagaag tgcgcctggc cagaaccatt gagaagcccg attctcctga tatgtgtatc
     1081 accaacgcaa gcagcattac cgactgtcat gttaatgtgt ctgttgtaga tctgcatgcg
     1141 gatccccctt cttccttgat cgacctgata ctcgcccagg ccaagactta tgtgaataac
     1201 cacgcttccg actacgtctg caaggtgatg gtgccacgtg gcgaatctat catcatgaac
     1261 aacacctacc cttacacccc tgagaggaaa gacgaaggga gaacaatcgt gccagtttct
     1321 gaatctcccc tgattagggc ccttatgagc atcgccaatc gtgtgagaat cgggtccatc
     1381 agattctttg tcgggagcca accccagcgg attagcattc tgatctccca cgttggtggc
     1441 gcccatgctg gttttactgg agacctggtg cctacaccag tgggtgaaag cgccgtcgac
     1501 tgggcccaag gactcgtgga tgcttatatc cagggcgtag tccctcaacc tctcccagtc
     1561 tatgggctcc ctggtgagat taaggggctg gtagctggac tgaacacatc ccagaccgta
     1621 tttgcgacac ttgacgtcga tgttagcatc gcagcttcca gcgctaactg ggtgaccatt
     1681 tacctgcagc cgggcgtaac cattgagaat ctccggattc aggcgaactc tgacggactc
     1741 ggcaatctcc tcacacacct cgcggcaccc gaggtggaga agcttatcaa cgctaagatc
     1801 gcgactgtgc tcgctgcact tgccgggtct gcgggcagcg gaccccacga caatcggacc
     1861 gccaacggta ccctgacatt ctcctttggg gagaacacag ttgtgcggga cagtcctctc
     1921 cggctgcccg gcgcgcccgg cagcctgcac cacatcctgg acgcccagaa gatgctgtgg
     1981 aaccacaggg acaggaacct gcctcctctg gcccctctgg gcccccacca ccatcaccac
     2041 cattgagtct agacacgtga ttaattaagg atcccccgac ctcgacctct ggctaataaa
     2101 ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga
     2161 catatgggag ggcaaatcat ttggtcgaga tccctcggag atctctagct agaggatcga
     2221 tccccgcccc ggacgaacta aacctgacta cgacatctct gccccttctt cgcggggcag
     2281 tgcatgtaat cccttcagtt ggttggtaca acttgccaac tgggccctgt tccacatgtg
     2341 acacgggggg ggaccaaaca caaaggggtt ctctgactgt agttgacatc cttataaatg
     2401 gatgtgcaca tttgccaaca ctgagtggct ttcatcctgg agcagacttt gcagtctgtg
     2461 gactgcaaca caacattgcc tttatgtgta actcttggct gaagctctta caccaatgct
     2521 gggggacatg tacctcccag gggcccagga agactacggg aggctacacc aacgtcaatc
     2581 agaggggcct gtgtagctac cgataagcgg accctcaaga gggcattagc aatagtgttt
     2641 ataaggcccc cttgttaacc ctaaacgggt agcatatgct tcccgggtag tagtatatac
     2701 tatccagact aaccctaatt caatagcata tgttacccaa cgggaagcat atgctatcga
     2761 attagggtta gtaaaagggt cctaaggaac agcgatatct cccaccccat gagctgtcac
     2821 ggttttattt acatggggtc aggattccac gagggtagtg aaccatttta gtcacaaggg
     2881 cagtggctga agatcaagga gcgggcagtg aactctcctg aatcttcgcc tgcttcttca
     2941 ttctccttcg tttagctaat agaataactg ctgagttgtg aacagtaagg tgtatgtgag
     3001 gtgctcgaaa acaaggtttc aggtgacgcc cccagaataa aatttggacg gggggttcag
     3061 tggtggcatt gtgctatgac accaatataa ccctcacaaa ccccttgggc aataaatact
     3121 agtgtaggaa tgaaacattc tgaatatctt taacaataga aatccatggg gtggggacaa
     3181 gccgtaaaga ctggatgtcc atctcacacg aatttatggc tatgggcaac acataatcct
     3241 agtgcaatat gatactgggg ttattaagat gtgtcccagg cagggaccaa gacaggtgaa
     3301 ccatgttgtt acactctatt tgtaacaagg ggaaagagag tggacgccga cagcagcgga
     3361 ctccactggt tgtctctaac acccccgaaa attaaacggg gctccacgcc aatggggccc
     3421 ataaacaaag acaagtggcc actctttttt ttgaaattgt ggagtggggg cacgcgtcag
     3481 cccccacacg ccgccctgcg gttttggact gtaaaataag ggtgtaataa cttggctgat
     3541 tgtaaccccg ctaaccactg cggtcaaacc acttgcccac aaaaccacta atggcacccc
     3601 ggggaatacc tgcataagta ggtgggcggg ccaagatagg ggcgcgattg ctgcgatctg
     3661 gaggacaaat tacacacact tgcgcctgag cgccaagcac agggttgttg gtcctcatat
     3721 tcacgaggtc gctgagagca cggtgggcta atgttgccat gggtagcata tactacccaa
     3781 atatctggat agcatatgct atcctaatct atatctgggt agcataggct atcctaatct
     3841 atatctgggt agcatatgct atcctaatct atatctgggt agtatatgct atcctaattt
     3901 atatctgggt agcataggct atcctaatct atatctgggt agcatatgct atcctaatct
     3961 atatctgggt agtatatgct atcctaatct gtatccgggt agcatatgct atcctaatag
     4021 agattagggt agtatatgct atcctaattt atatctgggt agcatatact acccaaatat
     4081 ctggatagca tatgctatcc taatctatat ctgggtagca tatgctatcc taatctatat
     4141 ctgggtagca taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat
     4201 ctgggtagta tatgctatcc taatttatat ctgggtagca taggctatcc taatctatat
     4261 ctgggtagca tatgctatcc taatctatat ctgggtagta tatgctatcc taatctgtat
     4321 ccgggtagca tatgctatcc tcatgataag ctgtcaaaca tgagaatttt cttgaagacg
     4381 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     4441 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4501 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4561 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     4621 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     4681 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     4741 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     4801 tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     4861 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     4921 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     4981 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     5041 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     5101 gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga
     5161 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     5221 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     5281 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     5341 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     5401 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     5461 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5521 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5581 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5641 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5701 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     5761 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5821 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5881 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     5941 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     6001 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6061 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6121 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6181 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     6241 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     6301 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     6361 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     6421 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     6481 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     6541 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
     6601 tgattacgcc aagctctagc tagaggtcga ccaattctca tgtttgacag cttatcatcg
     6661 cagatccggg caacgttgtt gccattgctg caggcgcaga actggtaggt atggaagatc
     6721 tatacattga atcaatattg gcaattagcc atattagtca ttggttatat agcataaatc
     6781 aatattggct attggccatt gcatacgttg tatctatatc ataatatgta catttatatt
     6841 ggctcatgtc caatatgacc gccatgttga cattgattat tgactagtta ttaatagtaa
     6901 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     6961 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     7021 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     7081 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtcc gccccctatt
     7141 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttacgggac
     7201 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatg
//
