LOCUS       sequence-262334-        7434 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 7434)
  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 7434)
  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: D41_Ld_180450
FEATURES             Location/Qualifiers
     source          1..7434
                     /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             2598..2615
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2667..2722
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2721..2740)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4958..4976)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5044..5148
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5149..5217,5218..6009)
                     /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(5367..5386)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6180..6768
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6669..6688
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6922..6939
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7056..7077
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7092..7097,7098..7115,7116..7122)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7130..7146
                     /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     7135..7157
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7154..7170
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7154..7170
                     /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 cctctacacc tcaccacggc tacgccagct gcgaccccag cgtggtgcag agcagcggct
     1261 acatcaacat ccccggcgtg aaccagaccc tgaagcacta cttctactgg ctgttcggcc
     1321 ccagaaagtg gcccaacgac ttccgcgagc cccccgtgat catgtggatg acaggcggcc
     1381 ctggctgcag cagctccatg gccctgctga ccgagctggg cccctgcatg atgaacgaga
     1441 caagcggcga gctgtaccac aacacctacg gctggaacga cgaggcctac ctgctgttcg
     1501 tggaccagcc caccggcgtg ggctacagct acggcgacaa gagcaactac gcccacaacc
     1561 agagcgaggt ggccgaggat atgtacaact tcctgcagct gttcgccaga cggttcacca
     1621 gccccagcat caccggcgcc aacgatttct acatcatcgg cgagtcctac ggcggccatt
     1681 acgtgcccgc cgtgtcccac agaatcctga tgggcaacga gcggagcgac ggcctgcaca
     1741 tcaacctgaa gggaatcgcc atcggcaacg gcctgaccga cccctacacc cagctgccct
     1801 tccacgccca gaccgcctac tactggtgca aagagaagct gggcgctccc tgtatcaccg
     1861 agaaggccta cgaggaaatg ctgagcctgc tgcccgcctg cctggaaaag accaagaagt
     1921 gcaacgaggg ccccgacgac agcgacgtgt cctgtagcgt ggccacagcc ctgtgggccg
     1981 agtatgtgga ccactactac gccaccggcc ggaactccta cgacatccgg aagcagtgta
     2041 tcggcgacct gtgctacccc atgcagaaca ccatcgactt ctaccacaag cctaccgtgc
     2101 gggccagcct gggagcctct gccaaggccc agtggtccac ctgtaacggc gaggtgtccg
     2161 ccctgttcga gcgggactac atgcggaact tcaacttcac cttcccccac atgctggaca
     2221 tgggcatccg ggtgctgatc tacgccggcg acatggactt catctgcaac tggctgggca
     2281 atgaggcctg ggtcaaggcc ctgcagtggt tcggcaccga cggcttcaac accgccccca
     2341 acgtggaatt tgccgtgtct ggcagatggg ccggacagga aagaagctac gggggcctga
     2401 gcttcgtgcg catctacgac gccggacaca tggtgcccat ggatcagcct gaggtggcac
     2461 tgttcatggt gcaccggttt ctgcacgacc ggcggctggc tggcgcgccc ggcagcctgc
     2521 accacatcct ggacgcccag aagatgctgt ggaaccacag ggacaggaac ctgcctcctc
     2581 tggcccctct gggcccccac caccatcacc accattgagt ctagacacgt gattaattaa
     2641 ggatcccccg acctcgacct ctggctaata aaggaaattt attttcattg caatagtgtg
     2701 ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc atttggtcga
     2761 gatccctcgg agatctctag ctagaggatc gatccccgcc ccggacgaac taaacctgac
     2821 tacgacatct ctgccccttc ttcgcggggc agtgcatgta atcccttcag ttggttggta
     2881 caacttgcca actgggccct gttccacatg tgacacgggg ggggaccaaa cacaaagggg
     2941 ttctctgact gtagttgaca tccttataaa tggatgtgca catttgccaa cactgagtgg
     3001 ctttcatcct ggagcagact ttgcagtctg tggactgcaa cacaacattg cctttatgtg
     3061 taactcttgg ctgaagctct tacaccaatg ctgggggaca tgtacctccc aggggcccag
     3121 gaagactacg ggaggctaca ccaacgtcaa tcagaggggc ctgtgtagct accgataagc
     3181 ggaccctcaa gagggcatta gcaatagtgt ttataaggcc cccttgttaa ccctaaacgg
     3241 gtagcatatg cttcccgggt agtagtatat actatccaga ctaaccctaa ttcaatagca
     3301 tatgttaccc aacgggaagc atatgctatc gaattagggt tagtaaaagg gtcctaagga
     3361 acagcgatat ctcccacccc atgagctgtc acggttttat ttacatgggg tcaggattcc
     3421 acgagggtag tgaaccattt tagtcacaag ggcagtggct gaagatcaag gagcgggcag
     3481 tgaactctcc tgaatcttcg cctgcttctt cattctcctt cgtttagcta atagaataac
     3541 tgctgagttg tgaacagtaa ggtgtatgtg aggtgctcga aaacaaggtt tcaggtgacg
     3601 cccccagaat aaaatttgga cggggggttc agtggtggca ttgtgctatg acaccaatat
     3661 aaccctcaca aaccccttgg gcaataaata ctagtgtagg aatgaaacat tctgaatatc
     3721 tttaacaata gaaatccatg gggtggggac aagccgtaaa gactggatgt ccatctcaca
     3781 cgaatttatg gctatgggca acacataatc ctagtgcaat atgatactgg ggttattaag
     3841 atgtgtccca ggcagggacc aagacaggtg aaccatgttg ttacactcta tttgtaacaa
     3901 ggggaaagag agtggacgcc gacagcagcg gactccactg gttgtctcta acacccccga
     3961 aaattaaacg gggctccacg ccaatggggc ccataaacaa agacaagtgg ccactctttt
     4021 ttttgaaatt gtggagtggg ggcacgcgtc agcccccaca cgccgccctg cggttttgga
     4081 ctgtaaaata agggtgtaat aacttggctg attgtaaccc cgctaaccac tgcggtcaaa
     4141 ccacttgccc acaaaaccac taatggcacc ccggggaata cctgcataag taggtgggcg
     4201 ggccaagata ggggcgcgat tgctgcgatc tggaggacaa attacacaca cttgcgcctg
     4261 agcgccaagc acagggttgt tggtcctcat attcacgagg tcgctgagag cacggtgggc
     4321 taatgttgcc atgggtagca tatactaccc aaatatctgg atagcatatg ctatcctaat
     4381 ctatatctgg gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat
     4441 ctatatctgg gtagtatatg ctatcctaat ttatatctgg gtagcatagg ctatcctaat
     4501 ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat
     4561 ctgtatccgg gtagcatatg ctatcctaat agagattagg gtagtatatg ctatcctaat
     4621 ttatatctgg gtagcatata ctacccaaat atctggatag catatgctat cctaatctat
     4681 atctgggtag catatgctat cctaatctat atctgggtag cataggctat cctaatctat
     4741 atctgggtag catatgctat cctaatctat atctgggtag tatatgctat cctaatttat
     4801 atctgggtag cataggctat cctaatctat atctgggtag catatgctat cctaatctat
     4861 atctgggtag tatatgctat cctaatctgt atccgggtag catatgctat cctcatgata
     4921 agctgtcaaa catgagaatt ttcttgaaga cgaaagggcc tcgtgatacg cctattttta
     4981 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     5041 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     5101 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     5161 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     5221 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     5281 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     5341 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc
     5401 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     5461 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     5521 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     5581 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     5641 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg
     5701 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     5761 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     5821 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     5881 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     5941 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     6001 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     6061 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     6121 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     6181 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     6241 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     6301 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     6361 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     6421 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     6481 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     6541 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     6601 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     6661 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     6721 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     6781 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     6841 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     6901 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     6961 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     7021 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     7081 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     7141 taacaatttc acacaggaaa cagctatgac catgattacg ccaagctcta gctagaggtc
     7201 gaccaattct catgtttgac agcttatcat cgcagatccg ggcaacgttg ttgccattgc
     7261 tgcaggcgca gaactggtag gtatggaaga tctatacatt gaatcaatat tggcaattag
     7321 ccatattagt cattggttat atagcataaa tcaatattgg ctattggcca ttgcatacgt
     7381 tgtatctata tcataatatg tacatttata ttggctcatg tccaatatga ccgc
//
