LOCUS       sequence-274907-        6846 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 6846)
  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 6846)
  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: D26_Ld_200260
FEATURES             Location/Qualifiers
     source          1..6846
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     2154..2173
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    2172..2227
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     CDS             complement(2279..2296)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     regulatory      3163..3172
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3195..3214)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     complement(3695..3719)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(3716..3919)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3745..3765)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3920..4299
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(4570..4586)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4570..4586)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4583..4605)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4594..4610
                     /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)."
     promoter        complement(join(4618..4624,4625..4642,4643..4648))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4663..4684
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4801..4818)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4972..5560)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5052..5071)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5731..6522,6523..6591))
                     /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     6354..6373
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6592..6696)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6764..6782
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gatacagatt aggatagcat atactaccca gatatagatt aggatagcat atgctaccca
       61 gatatagatt aggatagcct atgctaccca gatataaatt aggatagcat atactaccca
      121 gatatagatt aggatagcat atgctaccca gatatagatt aggatagcct atgctaccca
      181 gatatagatt aggatagcat atgctaccca gatatagatt aggatagcat atgctatcca
      241 gatatttggg tagtatatgc tacccagata taaattagga tagcatatac taccctaatc
      301 tctattagga tagcatatgc tacccggata cagattagga tagcatatac tacccagata
      361 tagattagga tagcatatgc tacccagata tagattagga tagcctatgc tacccagata
      421 taaattagga tagcatatac tacccagata tagattagga tagcatatgc tacccagata
      481 tagattagga tagcctatgc tacccagata tagattagga tagcatatgc tatccagata
      541 tttgggtagt atatgctacc catggcaaca ttagcccacc gtgctctcag cgacctcgtg
      601 aatatgagga ccaacaaccc tgtgcttggc gctcaggcgc aagtgtgtgt aatttgtcct
      661 ccagatcgca gcaatcgcgc ccctatcttg gcccgcccac ctacttatgc aggtattccc
      721 cggggtgcca ttagtggttt tgtgggcaag tggtttgacc gcagtggtta gcggggttac
      781 aatcagccaa gttattacac ccttatttta cagtccaaaa ccgcagggcg gcgtgtgggg
      841 gctgacgcgt gcccccactc cacaatttca aaaaaaagag tggccacttg tctttgttta
      901 tgggccccat tggcgtggag ccccgtttaa ttttcggggg tgttagagac aaccagtgga
      961 gtccgctgct gtcggcgtcc actctctttc cccttgttac aaatagagtg taacaacatg
     1021 gttcacctgt cttggtccct gcctgggaca catcttaata accccagtat catattgcac
     1081 taggattatg tgttgcccat agccataaat tcgtgtgaga tggacatcca gtctttacgg
     1141 cttgtcccca ccccatggat ttctattgtt aaagatattc agaatgtttc attcctacac
     1201 tagtatttat tgcccaaggg gtttgtgagg gttatattgg tgtcatagca caatgccacc
     1261 actgaacccc ccgtccaaat tttattctgg gggcgtcacc tgaaaccttg ttttcgagca
     1321 cctcacatac accttactgt tcacaactca gcagttattc tattagctaa acgaaggaga
     1381 atgaagaagc aggcgaagat tcaggagagt tcactgcccg ctccttgatc ttcagccact
     1441 gcccttgtga ctaaaatggt tcactaccct cgtggaatcc tgaccccatg taaataaaac
     1501 cgtgacagct catggggtgg gagatatcgc tgttccttag gaccctttta ctaaccctaa
     1561 ttcgatagca tatgcttccc gttgggtaac atatgctatt gaattagggt tagtctggat
     1621 agtatatact actacccggg aagcatatgc tacccgttta gggttaacaa gggggcctta
     1681 taaacactat tgctaatgcc ctcttgaggg tccgcttatc ggtagctaca caggcccctc
     1741 tgattgacgt tggtgtagcc tcccgtagtc ttcctgggcc cctgggaggt acatgtcccc
     1801 cagcattggt gtaagagctt cagccaagag ttacacataa aggcaatgtt gtgttgcagt
     1861 ccacagactg caaagtctgc tccaggatga aagccactca gtgttggcaa atgtgcacat
     1921 ccatttataa ggatgtcaac tacagtcaga gaaccccttt gtgtttggtc cccccccgtg
     1981 tcacatgtgg aacagggccc agttggcaag ttgtaccaac caactgaagg gattacatgc
     2041 actgccccgc gaagaagggg cagagatgtc gtagtcaggt ttagttcgtc cggggcgggg
     2101 atcgatcctc tagctagaga tctccgaggg atctcgacca aatgatttgc cctcccatat
     2161 gtccttccga gtgagagaca caaaaaattc caacacacta ttgcaatgaa aataaatttc
     2221 ctttattagc cagaggtcga ggtcggggga tccttaatta atcacgtgtc tagactcaat
     2281 ggtggtgatg gtggtggggg cccagagggg ccagaggagg caggttcctg tccctgtggt
     2341 tccacagcat cttctgggcg tccaggatgt ggtgcaggct gccgggcgcg cctcccttca
     2401 tgttctgcag cacggacatc acgatgggga tgaccaggta catccagtat ttctgcagga
     2461 aggaccgttc ctcgacgggc acttcctctt ccaccacttc gccctcggcg tttcggacca
     2521 tccgccgctc ggtctggggt ctggcgattg tctttctggc gtcctctgtc tcggtcaggg
     2581 cggtcaggtc ctcgtagcgg actttccggg tcactttcag agggtgcacc aggcccactt
     2641 tggtgttcag gtgcacggtg gggaacagtt tctgcaccac gctgcggtcg cactcgtcgc
     2701 ccttcagcat cttggcgtgg aagaagtttg tctcgctgga cacctggagg cccatcaggg
     2761 aggtggacac gccgggcacg atgttgatgt tctcgtgcag caccactgtc ttgctgtcca
     2821 tggcctcgaa gccccggatc aggctgcagg gggtgaccac cacggtggtg ctgtgcttgg
     2881 gggcaggctt gccttcctgg ttggggtagg cccggtagta caccaggtcg gcagcggcca
     2941 gcttgtttct ctcttccatg ctcagggaca cctcgccgcc cagctgtgta gagatccggg
     3001 caggggcctg gctgctcagt ctgctgatgg cgaagctgcc gatccgggac cactcgccgt
     3061 cgctgcccac tcttctttcc agcacgtagc tctgagacac ggcggtggcg gccgcaccag
     3121 acatgcagag cagcagggac atgagtacct gggtctgaaa ctccatggtg gcaaacttgg
     3181 acctgggagt ggacacctgt ggagagaaag gcaaagtgga tgtcattgtc actcaagtgt
     3241 atggccagat cgggccaggt gaatatcaaa tcctcctcgt ttttggaaac tgacaatctt
     3301 agcgcagaag taatgcccgc ttttgagagg gagtactcac cccaacagct ggatctcaag
     3361 cctgccacac ctcacctcga ccatccgccg tctcaagacc gcctacttta attacatcat
     3421 cagcagcacc tccgccagaa acaaccccga ccgccacccg ctgccgcccg ccacggtgct
     3481 cagcctacct tgcgactgtg actggttaga cgcctttctc gagaggtttt ccgatccggt
     3541 cgatgcggac tcgctcaggt ccctcggtgg cggagtaccg ttcggaggcc gacgggtttc
     3601 cgatccaaga gtactggaaa gaccgcgaag agtttgtcct caaccgcgag cccaacagct
     3661 ggccctcgca gacagcgatg cggaagagag tgaggatctg acggttcact aaacgagctc
     3721 tgcttatata gacctcccac cgtacacgcc taccgcccat ttgcgtcaac ggggcggggt
     3781 tattacgaca ttttggaaag tcccgttgat tttggtgcca aaacaaactc ccattgacgt
     3841 caatggggtg gagacttgga aatccccgtg agtcaaaccg ctatccacgc ccattggtgt
     3901 actgccaaaa ccgcatcacc atggtaatag cgatgactaa tacgtagatg tactgccaag
     3961 taggaaagtc ccgtaaggtc atgtactggg cataatgcca ggcgggccat ttaccgtcat
     4021 tgacgtcaat agggggcgga cttggcatat gatacacttg atgtactgcc aagtgggcag
     4081 tttaccgtaa atactccacc cattgacgtc aatggaaagt ccctattggc gttactatgg
     4141 gaacatacgt cattattgac gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc
     4201 catttaccgt aagttatgta acgcggaact ccatatatgg gctatgaact aatgaccccg
     4261 taattgatta ctattaataa ctagtcaata atcaatgtca acatggcggt catattggac
     4321 atgagccaat ataaatgtac atattatgat atagatacaa cgtatgcaat ggccaatagc
     4381 caatattgat ttatgctata taaccaatga ctaatatggc taattgccaa tattgattca
     4441 atgtatagat cttccatacc taccagttct gcgcctgcag caatggcaac aacgttgccc
     4501 ggatctgcga tgataagctg tcaaacatga gaattggtcg acctctagct agagcttggc
     4561 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     4621 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     4681 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     4741 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     4801 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     4861 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     4921 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4981 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5041 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5101 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5161 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5221 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5281 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5341 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5401 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5461 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5521 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5581 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     5641 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     5701 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     5761 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     5821 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     5881 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5941 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6001 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctgcag gcatcgtggt
     6061 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6121 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6181 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6241 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6301 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac gggataatac
     6361 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     6421 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6481 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6541 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     6601 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     6661 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     6721 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     6781 gccctttcgt cttcaagaaa attctcatgt ttgacagctt atcatgagga tagcatatgc
     6841 tacccg
//
