LOCUS       sequence-262383-        6324 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 6324)
  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 6324)
  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: D91_Ld_352550
FEATURES             Location/Qualifiers
     source          1..6324
                     /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             1488..1505
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1557..1612
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1611..1630)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3848..3866)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3934..4038
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4039..4107,4108..4899)
                     /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(4257..4276)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5070..5658
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5559..5578
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5812..5829
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5946..5967
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5982..5987,5988..6005,6006..6012)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6020..6036
                     /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     6025..6047
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6044..6060
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6044..6060
                     /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 gatttctgtc taagtttgga ctggccacac ctcaggctct gcaaggcgga attgctcctc
     1261 agcaggacac cgccttcaac gacatggtgc ctagcagctt cgacgccgac ttcagctcta
     1321 atggaaacgg cggcgtcaga tccgtgtcca gcatgaagct gcagatcgcc agcctgctgc
     1381 acagcatcag aggcgcgccc ggcagcctgc accacatcct ggacgcccag aagatgctgt
     1441 ggaaccacag ggacaggaac ctgcctcctc tggcccctct gggcccccac caccatcacc
     1501 accattgagt ctagacacgt gattaattaa ggatcccccg acctcgacct ctggctaata
     1561 aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag
     1621 gacatatggg agggcaaatc atttggtcga gatccctcgg agatctctag ctagaggatc
     1681 gatccccgcc ccggacgaac taaacctgac tacgacatct ctgccccttc ttcgcggggc
     1741 agtgcatgta atcccttcag ttggttggta caacttgcca actgggccct gttccacatg
     1801 tgacacgggg ggggaccaaa cacaaagggg ttctctgact gtagttgaca tccttataaa
     1861 tggatgtgca catttgccaa cactgagtgg ctttcatcct ggagcagact ttgcagtctg
     1921 tggactgcaa cacaacattg cctttatgtg taactcttgg ctgaagctct tacaccaatg
     1981 ctgggggaca tgtacctccc aggggcccag gaagactacg ggaggctaca ccaacgtcaa
     2041 tcagaggggc ctgtgtagct accgataagc ggaccctcaa gagggcatta gcaatagtgt
     2101 ttataaggcc cccttgttaa ccctaaacgg gtagcatatg cttcccgggt agtagtatat
     2161 actatccaga ctaaccctaa ttcaatagca tatgttaccc aacgggaagc atatgctatc
     2221 gaattagggt tagtaaaagg gtcctaagga acagcgatat ctcccacccc atgagctgtc
     2281 acggttttat ttacatgggg tcaggattcc acgagggtag tgaaccattt tagtcacaag
     2341 ggcagtggct gaagatcaag gagcgggcag tgaactctcc tgaatcttcg cctgcttctt
     2401 cattctcctt cgtttagcta atagaataac tgctgagttg tgaacagtaa ggtgtatgtg
     2461 aggtgctcga aaacaaggtt tcaggtgacg cccccagaat aaaatttgga cggggggttc
     2521 agtggtggca ttgtgctatg acaccaatat aaccctcaca aaccccttgg gcaataaata
     2581 ctagtgtagg aatgaaacat tctgaatatc tttaacaata gaaatccatg gggtggggac
     2641 aagccgtaaa gactggatgt ccatctcaca cgaatttatg gctatgggca acacataatc
     2701 ctagtgcaat atgatactgg ggttattaag atgtgtccca ggcagggacc aagacaggtg
     2761 aaccatgttg ttacactcta tttgtaacaa ggggaaagag agtggacgcc gacagcagcg
     2821 gactccactg gttgtctcta acacccccga aaattaaacg gggctccacg ccaatggggc
     2881 ccataaacaa agacaagtgg ccactctttt ttttgaaatt gtggagtggg ggcacgcgtc
     2941 agcccccaca cgccgccctg cggttttgga ctgtaaaata agggtgtaat aacttggctg
     3001 attgtaaccc cgctaaccac tgcggtcaaa ccacttgccc acaaaaccac taatggcacc
     3061 ccggggaata cctgcataag taggtgggcg ggccaagata ggggcgcgat tgctgcgatc
     3121 tggaggacaa attacacaca cttgcgcctg agcgccaagc acagggttgt tggtcctcat
     3181 attcacgagg tcgctgagag cacggtgggc taatgttgcc atgggtagca tatactaccc
     3241 aaatatctgg atagcatatg ctatcctaat ctatatctgg gtagcatagg ctatcctaat
     3301 ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat
     3361 ttatatctgg gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat
     3421 ctatatctgg gtagtatatg ctatcctaat ctgtatccgg gtagcatatg ctatcctaat
     3481 agagattagg gtagtatatg ctatcctaat ttatatctgg gtagcatata ctacccaaat
     3541 atctggatag catatgctat cctaatctat atctgggtag catatgctat cctaatctat
     3601 atctgggtag cataggctat cctaatctat atctgggtag catatgctat cctaatctat
     3661 atctgggtag tatatgctat cctaatttat atctgggtag cataggctat cctaatctat
     3721 atctgggtag catatgctat cctaatctat atctgggtag tatatgctat cctaatctgt
     3781 atccgggtag catatgctat cctcatgata agctgtcaaa catgagaatt ttcttgaaga
     3841 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     3901 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     3961 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4021 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     4081 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     4141 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     4201 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     4261 tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac
     4321 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     4381 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     4441 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     4501 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     4561 gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc
     4621 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     4681 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     4741 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     4801 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     4861 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     4921 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     4981 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5041 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5101 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5161 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     5221 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     5281 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     5341 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     5401 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     5461 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     5521 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     5581 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     5641 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     5701 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     5761 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     5821 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     5881 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     5941 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     6001 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     6061 catgattacg ccaagctcta gctagaggtc gaccaattct catgtttgac agcttatcat
     6121 cgcagatccg ggcaacgttg ttgccattgc tgcaggcgca gaactggtag gtatggaaga
     6181 tctatacatt gaatcaatat tggcaattag ccatattagt cattggttat atagcataaa
     6241 tcaatattgg ctattggcca ttgcatacgt tgtatctata tcataatatg tacatttata
     6301 ttggctcatg tccaatatga ccgc
//
