LOCUS       sequence-273278-        7752 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 7752)
  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 7752)
  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: D8_Ld_340400
FEATURES             Location/Qualifiers
     source          1..7752
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     201..225
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     706..725
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      748..757
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2530..2547
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2599..2654
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2653..2672)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4890..4908)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4976..5080
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5081..5149,5150..5941)
                     /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(5299..5318)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6112..6700
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6601..6620
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6854..6871
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6988..7009
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7024..7029,7030..7047,7048..7054)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7062..7078
                     /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     7067..7089
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7086..7102
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7086..7102
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7373..7752
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta caccaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcctcac tctcttccgc
      241 atcgctgtct gcgagggcca gctgttgggc tcgcggttga ggacaaactc ttcgcggtct
      301 ttccagtact cttggatcgg aaacccgtcg gcctccgaac ggtactccgc caccgaggga
      361 cctgagcgag tccgcatcga ccggatcgga aaacctctcg agaaaggcgt ctaaccagtc
      421 acagtcgcaa ggtaggctga gcaccgtggc gggcggcagc gggtggcggt cggggttgtt
      481 tctggcggag gtgctgctga tgatgtaatt aaagtaggcg gtcttgagac ggcggatggt
      541 cgaggtgagg tgtggcaggc ttgagatcca gctgttgggg tgagtactcc ctctcaaaag
      601 cgggcattac ttctgcgcta agattgtcag tttccaaaaa cgaggaggat ttgatattca
      661 cctggcccga tctggccata cacttgagtg acaatgacat ccactttgcc tttctctcca
      721 caggtgtcca ctcccaggtc caagtttgcc accatggagt ttcagaccca ggtactcatg
      781 tccctgctgc tctgcatgtc tggtgcggcc gccaccctgt ctgccgtgga tcctgatgcc
      841 ggcgctgacg ccaacggcag atctagccct gcccctgcct ctgccgtggc cggcgagcac
      901 gtgaccgtgt tcatgtgcgc cggcagcagc gacttcagcg gcgtgcacga gctgggcggc
      961 ctgatgaagg ccatggaaga gacacaggtg ctgagcctga agtactgcca gaccgcccac
     1021 ctgggctaca tcagacacgc cgatggacac gccgtgcgga gcctggtggt caccaccggc
     1081 atcaactaca tcagcagcac cctgtgcacc agaagcgtgc tgaagctgcg gcggcagcgg
     1141 ggctacacct acgacgccat ggtgttcatc ggcaccagcg gctttagccc catggtcgga
     1201 ggctgggacc ccaccaacac aagcagctac gccgccagca aggccagcga gcaggaagcc
     1261 atgatgaagg gcgaagccgc cacagccggc gtgggagctg ctcctcctgc cctgccactg
     1321 aaaaccctgc agcaggtccg agaggaacag gccgaggaag cccagctgag acaggccgct
     1381 gtggacggcg gccagctgtt taccctgcct accgccagcc agctgaactt cgagaacgcc
     1441 cggaccggcg aacaggtgga ctgggacggc tgcagcccta gagtgcccag cgccatcaca
     1501 cccctgagcc tgggcagcgt gtgtgtgacc agcaccgcct acctgatgga aaccggctgc
     1561 tgcaccgagc gcgtgcggca cagccagtgc agcagacccc actgcaccag cttccgcagc
     1621 gagctgggct ccgtgaccaa ggtgttcttc accagcgacg ccctggcccg gaagatcgag
     1681 gccgcctctc accacaaggc ctggcccgcc atgcccgaga tcgtgaagaa aggccagatg
     1741 cggttctggg ccaccaacga ggccgtggaa cctgtgggcg agggcgccga tagacacaga
     1801 gccgccccta gcggccccag cttcgtgaca tgtgccgagg ccaccggcaa caccatcaac
     1861 acaggcgccg agcgggacta cctgtgcaga gagcacacca cccaggccct ggacatggcc
     1921 cagtacagac tggccgagct gcaggacggc aaggccgctc ctcaggaacc ccccctgacc
     1981 acccagaatg tggcctgcgt gcgggctatg gaatccatgg gctttctgca cagcgtggcc
     2041 gaggacagca ccgccagaga catccctgtg gccgtcgtgc ggggagccgc caactacgac
     2101 atgtacccca tgaagaaagt gtacgtgccc cctgccgtgt ggcggagcgc tctggtggct
     2161 ggcagactga aggtgtcccc cagagtggcc gctgccctga aggccgaaga ggccgctgct
     2221 gccaatggcg agggggccct gagagagagc gccattgccg cctacacctg gcaccagaat
     2281 gtgcacttca tgagcgaggc cgagcaccag gccttcgtga acgccagctt ccactacgcc
     2341 gtggaaaccg ccaccttcgt gctgagcaac tacttcttcg gcggcagatg cttcgccagc
     2401 gccagcggag ccagacccgc cagaagcacc gaaggcgcgc ccggcagcct gcaccacatc
     2461 ctggacgccc agaagatgct gtggaaccac agggacagga acctgcctcc tctggcccct
     2521 ctgggccccc accaccatca ccaccattga gtctagacac gtgattaatt aaggatcccc
     2581 cgacctcgac ctctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt
     2641 ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttggtc gagatccctc
     2701 ggagatctct agctagagga tcgatccccg ccccggacga actaaacctg actacgacat
     2761 ctctgcccct tcttcgcggg gcagtgcatg taatcccttc agttggttgg tacaacttgc
     2821 caactgggcc ctgttccaca tgtgacacgg ggggggacca aacacaaagg ggttctctga
     2881 ctgtagttga catccttata aatggatgtg cacatttgcc aacactgagt ggctttcatc
     2941 ctggagcaga ctttgcagtc tgtggactgc aacacaacat tgcctttatg tgtaactctt
     3001 ggctgaagct cttacaccaa tgctggggga catgtacctc ccaggggccc aggaagacta
     3061 cgggaggcta caccaacgtc aatcagaggg gcctgtgtag ctaccgataa gcggaccctc
     3121 aagagggcat tagcaatagt gtttataagg cccccttgtt aaccctaaac gggtagcata
     3181 tgcttcccgg gtagtagtat atactatcca gactaaccct aattcaatag catatgttac
     3241 ccaacgggaa gcatatgcta tcgaattagg gttagtaaaa gggtcctaag gaacagcgat
     3301 atctcccacc ccatgagctg tcacggtttt atttacatgg ggtcaggatt ccacgagggt
     3361 agtgaaccat tttagtcaca agggcagtgg ctgaagatca aggagcgggc agtgaactct
     3421 cctgaatctt cgcctgcttc ttcattctcc ttcgtttagc taatagaata actgctgagt
     3481 tgtgaacagt aaggtgtatg tgaggtgctc gaaaacaagg tttcaggtga cgcccccaga
     3541 ataaaatttg gacggggggt tcagtggtgg cattgtgcta tgacaccaat ataaccctca
     3601 caaacccctt gggcaataaa tactagtgta ggaatgaaac attctgaata tctttaacaa
     3661 tagaaatcca tggggtgggg acaagccgta aagactggat gtccatctca cacgaattta
     3721 tggctatggg caacacataa tcctagtgca atatgatact ggggttatta agatgtgtcc
     3781 caggcaggga ccaagacagg tgaaccatgt tgttacactc tatttgtaac aaggggaaag
     3841 agagtggacg ccgacagcag cggactccac tggttgtctc taacaccccc gaaaattaaa
     3901 cggggctcca cgccaatggg gcccataaac aaagacaagt ggccactctt ttttttgaaa
     3961 ttgtggagtg ggggcacgcg tcagccccca cacgccgccc tgcggttttg gactgtaaaa
     4021 taagggtgta ataacttggc tgattgtaac cccgctaacc actgcggtca aaccacttgc
     4081 ccacaaaacc actaatggca ccccggggaa tacctgcata agtaggtggg cgggccaaga
     4141 taggggcgcg attgctgcga tctggaggac aaattacaca cacttgcgcc tgagcgccaa
     4201 gcacagggtt gttggtcctc atattcacga ggtcgctgag agcacggtgg gctaatgttg
     4261 ccatgggtag catatactac ccaaatatct ggatagcata tgctatccta atctatatct
     4321 gggtagcata ggctatccta atctatatct gggtagcata tgctatccta atctatatct
     4381 gggtagtata tgctatccta atttatatct gggtagcata ggctatccta atctatatct
     4441 gggtagcata tgctatccta atctatatct gggtagtata tgctatccta atctgtatcc
     4501 gggtagcata tgctatccta atagagatta gggtagtata tgctatccta atttatatct
     4561 gggtagcata tactacccaa atatctggat agcatatgct atcctaatct atatctgggt
     4621 agcatatgct atcctaatct atatctgggt agcataggct atcctaatct atatctgggt
     4681 agcatatgct atcctaatct atatctgggt agtatatgct atcctaattt atatctgggt
     4741 agcataggct atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt
     4801 agtatatgct atcctaatct gtatccgggt agcatatgct atcctcatga taagctgtca
     4861 aacatgagaa ttttcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     4921 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     4981 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     5041 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     5101 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     5161 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     5221 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     5281 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga
     5341 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     5401 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     5461 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     5521 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     5581 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac
     5641 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     5701 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     5761 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     5821 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     5881 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     5941 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     6001 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     6061 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     6121 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     6181 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     6241 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     6301 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     6361 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     6421 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     6481 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     6541 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     6601 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     6661 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     6721 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     6781 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     6841 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     6901 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     6961 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
     7021 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     7081 tcacacagga aacagctatg accatgatta cgccaagctc tagctagagg tcgaccaatt
     7141 ctcatgtttg acagcttatc atcgcagatc cgggcaacgt tgttgccatt gctgcaggcg
     7201 cagaactggt aggtatggaa gatctataca ttgaatcaat attggcaatt agccatatta
     7261 gtcattggtt atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta
     7321 tatcataata tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga
     7381 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     7441 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     7501 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     7561 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     7621 catatgccaa gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     7681 gcccagtaca tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc
     7741 gctattacca tg
//
