LOCUS       sequence-274655-        8310 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 8310)
  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 8310)
  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: D51_Ld_310120
FEATURES             Location/Qualifiers
     source          1..8310
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     1086..1105
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1128..1137
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3468..3485
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3537..3592
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3591..3610)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5828..5846)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5914..6018
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6019..6087,6088..6879)
                     /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(6237..6256)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7050..7638
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7539..7558
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7792..7809
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7926..7947
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7962..7967,7968..7985,7986..7992)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8000..8016
                     /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     8005..8027
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8024..8040
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8024..8040
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agatcctcac tctcttccgc atcgctgtct gcgagggcca gctgttgggc tcgcggttga
      661 ggacaaactc ttcgcggtct ttccagtact cttggatcgg aaacccgtcg gcctccgaac
      721 ggtactccgc caccgaggga cctgagcgag tccgcatcga ccggatcgga aaacctctcg
      781 agaaaggcgt ctaaccagtc acagtcgcaa ggtaggctga gcaccgtggc gggcggcagc
      841 gggtggcggt cggggttgtt tctggcggag gtgctgctga tgatgtaatt aaagtaggcg
      901 gtcttgagac ggcggatggt cgaggtgagg tgtggcaggc ttgagatcca gctgttgggg
      961 tgagtactcc ctctcaaaag cgggcattac ttctgcgcta agattgtcag tttccaaaaa
     1021 cgaggaggat ttgatattca cctggcccga tctggccata cacttgagtg acaatgacat
     1081 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caagtttgcc accatggagt
     1141 ttcagaccca ggtactcatg tccctgctgc tctgcatgtc tggtgcggcc gcccaggatg
     1201 tggaacagct ggtgcccaac agcaccagcc agtgctaccc cagcatcgac ctggggtggt
     1261 ccgcccacgt gggaagcagc gtgtacgcca cccccagaat cgtggacctg gcccacgacg
     1321 gccacaaaga ggtgctgatc cctaccttca gccagttcct ggaagccctg gacggcccca
     1381 ccggcgagga tatccccggc ttcccattca tgcacccccg gatgaagacc tacgccagcc
     1441 ccctgcccgt ggacctgaat ggcgacggac aggtggactg gctggtggcc ctgtacagcg
     1501 gcgagctggt ggtgttcggc caggacggca aggtgcaggg ctacctccag gtgccccccc
     1561 tggccatgaa gcggaactgg gtcaaggaca acctgccccc ccagcaggcc gccacagtca
     1621 gacctggcac ccccctgaaa accggcaccg tgctggaaga tatcctgaag gccagacgga
     1681 tggacgacct gaccctgacc agcaccgagg tgctggacca gcggcggaga agcagcatcg
     1741 tgcagagaca ccacgccccc accgccgctg ccgtgaagtc cgtggaatac gacacccacc
     1801 ccaagggcgc cgagctgaga gccaccgagg acaagcccga ccagtggaac ggcgacgacg
     1861 ccctggacat cgacgacgac gagatggaac tgccctggga gaaggccgag gtggaagtgg
     1921 gccccaacag ccacctgagc cctgaggcca gagccagcct ggacctgctg ttccaccccg
     1981 agctgtacca gaccgccaag agagccacaa gcgccgacga ggccaccttc gagggccgga
     2041 atctgcggca ggccacctct atcgccttcg gcgaggacga gatcaaggtg gacgcccaca
     2101 tcctgagcac ccccgtggtc accgacgtgg accagaacgg cgatctggac gtgatcctgc
     2161 acgtgtccta ctttttcgga gaggacgtga tcgccgagca gcccgagaga ttcgctggcg
     2221 gcgtggaccc ccacgatttc gtggccacag ccgtcgtgtg cctgaacctg gtcaccggcg
     2281 agacaagatg ggccagactg ctgcacgcca gcaccaagaa cagcaccaac gccgcctacg
     2341 ccctgtccac ccccctggtc atcaacgccg atgaggaccg gtccatggaa gtgttcgtgt
     2401 ccaccagcct gggcttcgtg ttcggcttca ccgccgacgg caacaccctg gatggctggc
     2461 ctgtgctgct gggccctctg agcgccagcc ctaccgccga agatatcaca ggcgacggca
     2521 ccctggatat ctgcatcggc gacaccgagg gcaaagtgcg gtgctacaac gccagcggca
     2581 gcgtgaccct ggaaaaggat gtgatcggcg gcgtggccga ccggctgacc ttcggagatg
     2641 tggatggcga cggcgtggtg gacatcgtgt ttggcaccac cagcggcctg atctacgccc
     2701 tgagaggcac cgatggcgcc gtgctgccca gcttccctat cgtgaccggc ggctctatcg
     2761 tggcccctgc cctgctgctg aacctgaacg gcatgctgta ctccaacgac ctggatatcg
     2821 tggtgcccag ccacaacggc cgggtgtacc tgctgagagg cagcagcggc tgcgtgcaga
     2881 ccatcgacat cgatgagaag tccagcagca tggtgctggc cgacgacgtg ctgggcaacg
     2941 gccagatgga cctggtggtc accaccatca agggcggcgt gtacgtgttc cagaccagca
     3001 ccccctacac cgccatgaac gcctggccca gcaagaccaa gggcgtgaac ggcttcagcg
     3061 cctccggcaa ctacatcgga atcgccatca gccccaactt ccggaccttc cgggacatcc
     3121 agggcgacac cttcaacgtg gaattcacca tctacgacag cagacccgtg tcccccaagc
     3181 ggagctacac cgtggaaatc accgtgggca cccggatccg gctgtacaga gagcggttcc
     3241 acaagcccgg cacccacatc gtgaaagtgc gggcacccct ggaacggatg tacagcagcc
     3301 tgaacgtggc cctgaccctg cccaacggcc agtccttcac cgacagcatc agcctgagct
     3361 tcaacatgca tggcgcgccc ggcagcctgc accacatcct ggacgcccag aagatgctgt
     3421 ggaaccacag ggacaggaac ctgcctcctc tggcccctct gggcccccac caccatcacc
     3481 accattgagt ctagacacgt gattaattaa ggatcccccg acctcgacct ctggctaata
     3541 aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag
     3601 gacatatggg agggcaaatc atttggtcga gatccctcgg agatctctag ctagaggatc
     3661 gatccccgcc ccggacgaac taaacctgac tacgacatct ctgccccttc ttcgcggggc
     3721 agtgcatgta atcccttcag ttggttggta caacttgcca actgggccct gttccacatg
     3781 tgacacgggg ggggaccaaa cacaaagggg ttctctgact gtagttgaca tccttataaa
     3841 tggatgtgca catttgccaa cactgagtgg ctttcatcct ggagcagact ttgcagtctg
     3901 tggactgcaa cacaacattg cctttatgtg taactcttgg ctgaagctct tacaccaatg
     3961 ctgggggaca tgtacctccc aggggcccag gaagactacg ggaggctaca ccaacgtcaa
     4021 tcagaggggc ctgtgtagct accgataagc ggaccctcaa gagggcatta gcaatagtgt
     4081 ttataaggcc cccttgttaa ccctaaacgg gtagcatatg cttcccgggt agtagtatat
     4141 actatccaga ctaaccctaa ttcaatagca tatgttaccc aacgggaagc atatgctatc
     4201 gaattagggt tagtaaaagg gtcctaagga acagcgatat ctcccacccc atgagctgtc
     4261 acggttttat ttacatgggg tcaggattcc acgagggtag tgaaccattt tagtcacaag
     4321 ggcagtggct gaagatcaag gagcgggcag tgaactctcc tgaatcttcg cctgcttctt
     4381 cattctcctt cgtttagcta atagaataac tgctgagttg tgaacagtaa ggtgtatgtg
     4441 aggtgctcga aaacaaggtt tcaggtgacg cccccagaat aaaatttgga cggggggttc
     4501 agtggtggca ttgtgctatg acaccaatat aaccctcaca aaccccttgg gcaataaata
     4561 ctagtgtagg aatgaaacat tctgaatatc tttaacaata gaaatccatg gggtggggac
     4621 aagccgtaaa gactggatgt ccatctcaca cgaatttatg gctatgggca acacataatc
     4681 ctagtgcaat atgatactgg ggttattaag atgtgtccca ggcagggacc aagacaggtg
     4741 aaccatgttg ttacactcta tttgtaacaa ggggaaagag agtggacgcc gacagcagcg
     4801 gactccactg gttgtctcta acacccccga aaattaaacg gggctccacg ccaatggggc
     4861 ccataaacaa agacaagtgg ccactctttt ttttgaaatt gtggagtggg ggcacgcgtc
     4921 agcccccaca cgccgccctg cggttttgga ctgtaaaata agggtgtaat aacttggctg
     4981 attgtaaccc cgctaaccac tgcggtcaaa ccacttgccc acaaaaccac taatggcacc
     5041 ccggggaata cctgcataag taggtgggcg ggccaagata ggggcgcgat tgctgcgatc
     5101 tggaggacaa attacacaca cttgcgcctg agcgccaagc acagggttgt tggtcctcat
     5161 attcacgagg tcgctgagag cacggtgggc taatgttgcc atgggtagca tatactaccc
     5221 aaatatctgg atagcatatg ctatcctaat ctatatctgg gtagcatagg ctatcctaat
     5281 ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat
     5341 ttatatctgg gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat
     5401 ctatatctgg gtagtatatg ctatcctaat ctgtatccgg gtagcatatg ctatcctaat
     5461 agagattagg gtagtatatg ctatcctaat ttatatctgg gtagcatata ctacccaaat
     5521 atctggatag catatgctat cctaatctat atctgggtag catatgctat cctaatctat
     5581 atctgggtag cataggctat cctaatctat atctgggtag catatgctat cctaatctat
     5641 atctgggtag tatatgctat cctaatttat atctgggtag cataggctat cctaatctat
     5701 atctgggtag catatgctat cctaatctat atctgggtag tatatgctat cctaatctgt
     5761 atccgggtag catatgctat cctcatgata agctgtcaaa catgagaatt ttcttgaaga
     5821 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     5881 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     5941 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     6001 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     6061 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     6121 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     6181 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     6241 tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac
     6301 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     6361 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     6421 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     6481 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     6541 gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc
     6601 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     6661 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     6721 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     6781 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     6841 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     6901 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     6961 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     7021 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     7081 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     7141 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     7201 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     7261 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     7321 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     7381 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     7441 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     7501 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     7561 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     7621 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     7681 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     7741 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     7801 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     7861 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     7921 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     7981 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     8041 catgattacg ccaagctcta gctagaggtc gaccaattct catgtttgac agcttatcat
     8101 cgcagatccg ggcaacgttg ttgccattgc tgcaggcgca gaactggtag gtatggaaga
     8161 tctatacatt gaatcaatat tggcaattag ccatattagt cattggttat atagcataaa
     8221 tcaatattgg ctattggcca ttgcatacgt tgtatctata tcataatatg tacatttata
     8281 ttggctcatg tccaatatga ccgccatgtt
//
