LOCUS       sequence-262299-        8376 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 8376)
  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 8376)
  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: D4__Ld_290790
FEATURES             Location/Qualifiers
     source          1..8376
                     /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             3540..3557
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3609..3664
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3663..3682)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5900..5918)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5986..6090
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6091..6159,6160..6951)
                     /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(6309..6328)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7122..7710
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7611..7630
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7864..7881
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7998..8019
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8034..8039,8040..8057,8058..8064)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8072..8088
                     /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     8077..8099
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8096..8112
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8096..8112
                     /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 ccggggatgg cagaggcacc ccaatcgcat tccaggccga ggtgtccaag atgctggaca
     1261 tcctggtcaa cagcctgtat accaaccggg ccgtgttcct gagagagctg atcagcaacg
     1321 gcagcgacgc cctggacaag atccgggtgc tgtacctgac cagccccaaa gagcccctga
     1381 ccaaggacgg cgaggccccc accatggacc tgcggatcag cttcgacaaa gagaagtccg
     1441 agctgatcct gcgggacggc ggcgtgggca tgaccaaaga ggaactggcc aagcacctgg
     1501 gcagcctggg caccagcggc accaagcact ttctggaaaa gctgcaggaa ggcgtcggcg
     1561 ccgtcggagg cgaccagaac aatctgatcg gccagttcgg cgtgggcttc tacagcgtgt
     1621 tcctcgtggg cgacagagtg cgggtggcca gcaagagcga cgacagcgac gagcagtacg
     1681 tgtgggagag caagggcgac ggccagtact tcctgtaccc cgaccccaga ggcaacaccc
     1741 tgggcagggg caccgagatc accatcgagc tgaagcccga cgccgagcag ttcctgagcg
     1801 ccgagacaat caagaaaacc atccaccagt acagcgagtt catcaacttc cccatctacg
     1861 tgcaggaaga ggtcgaggtc gccagcaccg ccgccacacc tgagtctgcc gccgaggaaa
     1921 gaagcctgga cgagggcgcc gtggaagagg accccgacaa agagggcgac acccagggcg
     1981 tggtcaaaga aaagcggtgg accctggtca acgagaaccg gcccatctgg acccggccca
     2041 tcggcaacgt gaccgaggaa gagtaccaca agttctacaa ggccttcagc ggcgactacc
     2101 gggaccccct gtacttcaac cacttcaagg tggaaggcga ggtggacttc gacagcatcc
     2161 tgttcgtgcc caccaccgtg gaccccgcca gcttcagcga cgacaacgcc gtgcccaaca
     2221 ccaacatcaa gctgtacgtg cggcgggtgt tcatcaccga cgagttccgg gacctgctgc
     2281 ccagatacct gaacttcgtg aagggcatcg tggacagcaa cgacctgccc ctgaacgtgt
     2341 ccagagaggt gctgcaggaa agccggatcc tgagagtgat caagaaaaag ctcgtgcgca
     2401 agaccctgag catgttcgcc gatatcgccg cccaggacga ggccattgcc aacggcaaac
     2461 aggtggaaaa ccctgccccc agcggccaca cccacctgaa gaagcccgcc tacaccaagt
     2521 tctgggagct gtacggcaag cacctgagac tgggcgtgat gctggactcc aacaaccgga
     2581 accggctgac caagctgttc cggtacaaga gcagcagaag cgagagcgag tacatcagcc
     2641 tgcagaccta cgtggaccgg atgaagaagg gccagaaggg catctactac ctgagcggcg
     2701 acagcgtggc ccggatcaag aaatcccccg tgctggaaga tgccgtgaac cacgacgtgg
     2761 aagtgatctt catgaccgac gccatcgacg agtacgtggt gtcccagctg accgacttcg
     2821 ccggcaagaa gctgatcaac ctggccaaag aaggcgtgca gttcgaggaa agcgacgccc
     2881 ggcagcgggt ggtggaccgg aagcggaaag agaagtacga cagcttcttt acccacctga
     2941 gagccctgtt cggctacagc gaagtgcgga aagtgatcct gaccaagcgg atgaccaacg
     3001 aggccttcat cgtgtccagc agcgagaacc agatcaccgc cagactggcc agcatcatgc
     3061 ggggccagag catgagcctg gccaaccagc agatgaccgc cgagcgggtg ctggaagtga
     3121 actacagaca ccccctggtg gacgagatgt tcaagcggtt caccgtggac gaggacgacg
     3181 aggtggccac cgacattgcc tgggtgctgt atgacaccgc caacctgcag gccgagttcc
     3241 ccgtggccga tgtggccgcc tacagcaagc ggatcaaccg gctgctgcgg agcagcgtgg
     3301 acctgtccgc cgacgatagc ctgctgcccc ccgacgatgc cgagtacacc gtgtccgaca
     3361 ccgaaaccga agaggaagag gaacagccca aggtggacac caacgcccac gaagaggccg
     3421 agacagacgg cgagggagac ctgggcgcgc ccggcagcct gcaccacatc ctggacgccc
     3481 agaagatgct gtggaaccac agggacagga acctgcctcc tctggcccct ctgggccccc
     3541 accaccatca ccaccattga gtctagacac gtgattaatt aaggatcccc cgacctcgac
     3601 ctctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct
     3661 ctcactcgga aggacatatg ggagggcaaa tcatttggtc gagatccctc ggagatctct
     3721 agctagagga tcgatccccg ccccggacga actaaacctg actacgacat ctctgcccct
     3781 tcttcgcggg gcagtgcatg taatcccttc agttggttgg tacaacttgc caactgggcc
     3841 ctgttccaca tgtgacacgg ggggggacca aacacaaagg ggttctctga ctgtagttga
     3901 catccttata aatggatgtg cacatttgcc aacactgagt ggctttcatc ctggagcaga
     3961 ctttgcagtc tgtggactgc aacacaacat tgcctttatg tgtaactctt ggctgaagct
     4021 cttacaccaa tgctggggga catgtacctc ccaggggccc aggaagacta cgggaggcta
     4081 caccaacgtc aatcagaggg gcctgtgtag ctaccgataa gcggaccctc aagagggcat
     4141 tagcaatagt gtttataagg cccccttgtt aaccctaaac gggtagcata tgcttcccgg
     4201 gtagtagtat atactatcca gactaaccct aattcaatag catatgttac ccaacgggaa
     4261 gcatatgcta tcgaattagg gttagtaaaa gggtcctaag gaacagcgat atctcccacc
     4321 ccatgagctg tcacggtttt atttacatgg ggtcaggatt ccacgagggt agtgaaccat
     4381 tttagtcaca agggcagtgg ctgaagatca aggagcgggc agtgaactct cctgaatctt
     4441 cgcctgcttc ttcattctcc ttcgtttagc taatagaata actgctgagt tgtgaacagt
     4501 aaggtgtatg tgaggtgctc gaaaacaagg tttcaggtga cgcccccaga ataaaatttg
     4561 gacggggggt tcagtggtgg cattgtgcta tgacaccaat ataaccctca caaacccctt
     4621 gggcaataaa tactagtgta ggaatgaaac attctgaata tctttaacaa tagaaatcca
     4681 tggggtgggg acaagccgta aagactggat gtccatctca cacgaattta tggctatggg
     4741 caacacataa tcctagtgca atatgatact ggggttatta agatgtgtcc caggcaggga
     4801 ccaagacagg tgaaccatgt tgttacactc tatttgtaac aaggggaaag agagtggacg
     4861 ccgacagcag cggactccac tggttgtctc taacaccccc gaaaattaaa cggggctcca
     4921 cgccaatggg gcccataaac aaagacaagt ggccactctt ttttttgaaa ttgtggagtg
     4981 ggggcacgcg tcagccccca cacgccgccc tgcggttttg gactgtaaaa taagggtgta
     5041 ataacttggc tgattgtaac cccgctaacc actgcggtca aaccacttgc ccacaaaacc
     5101 actaatggca ccccggggaa tacctgcata agtaggtggg cgggccaaga taggggcgcg
     5161 attgctgcga tctggaggac aaattacaca cacttgcgcc tgagcgccaa gcacagggtt
     5221 gttggtcctc atattcacga ggtcgctgag agcacggtgg gctaatgttg ccatgggtag
     5281 catatactac ccaaatatct ggatagcata tgctatccta atctatatct gggtagcata
     5341 ggctatccta atctatatct gggtagcata tgctatccta atctatatct gggtagtata
     5401 tgctatccta atttatatct gggtagcata ggctatccta atctatatct gggtagcata
     5461 tgctatccta atctatatct gggtagtata tgctatccta atctgtatcc gggtagcata
     5521 tgctatccta atagagatta gggtagtata tgctatccta atttatatct gggtagcata
     5581 tactacccaa atatctggat agcatatgct atcctaatct atatctgggt agcatatgct
     5641 atcctaatct atatctgggt agcataggct atcctaatct atatctgggt agcatatgct
     5701 atcctaatct atatctgggt agtatatgct atcctaattt atatctgggt agcataggct
     5761 atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt agtatatgct
     5821 atcctaatct gtatccgggt agcatatgct atcctcatga taagctgtca aacatgagaa
     5881 ttttcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     5941 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     6001 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     6061 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     6121 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     6181 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     6241 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     6301 aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt
     6361 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     6421 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     6481 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     6541 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     6601 ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa
     6661 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     6721 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     6781 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     6841 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     6901 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     6961 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     7021 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     7081 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     7141 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     7201 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     7261 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     7321 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     7381 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     7441 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     7501 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     7561 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     7621 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     7681 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     7741 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     7801 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     7861 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     7921 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     7981 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     8041 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     8101 aacagctatg accatgatta cgccaagctc tagctagagg tcgaccaatt ctcatgtttg
     8161 acagcttatc atcgcagatc cgggcaacgt tgttgccatt gctgcaggcg cagaactggt
     8221 aggtatggaa gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt
     8281 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata
     8341 tgtacattta tattggctca tgtccaatat gaccgc
//
