LOCUS       sequence-274666-        7272 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 7272)
  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 7272)
  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: D93_Ld_361080
FEATURES             Location/Qualifiers
     source          1..7272
                     /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             2050..2067
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2119..2174
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2173..2192)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4410..4428)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4496..4600
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4601..4669,4670..5461)
                     /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(4819..4838)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5632..6220
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6121..6140
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6374..6391
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6508..6529
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6544..6549,6550..6567,6568..6574)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6582..6598
                     /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     6587..6609
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6606..6622
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6606..6622
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        6893..7272
                     /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 gccgatactg ctcatcctca ccattctagg
      841 tcccatacag tcacaataga gtccggaaca ctgcccgaag caaaggcttt tgcgcccttg
      901 aggctgcggg ccggggaatg ggtatataac ttcaatccag gcgaagaagc attcttcttt
      961 gagcccgagg atgccggtaa caggcagtgg cagcagtgga aggctgatct caggttgttg
     1021 caggcctatc aggagaaaca gcgactttcc aactatccct ctggcgggga gaagactgac
     1081 acccccgata agcctggaaa tactgagcgc agagagccgg gcgcgttgcc tattgccgag
     1141 tatttctccg accgcaacgt tcagaggcgg tatgctgaac gcctgaagga acagtctttt
     1201 cataaccttt ccgacgcaca gggtgctgca gatgctcgcg ccctggtaga aggcgagata
     1261 gctgcaatgt ggcatctcgt cagaagtgct ggcagttcag atagccttaa tgatcagcag
     1321 tttctgggac tggtggaaag gcatttcagt catactatag ctaatacctc agcagctgcc
     1381 gcagccgaag ccacaacacc ttcaagcttt cccttcgcct tgcttcgcgt tcccctggta
     1441 ctcgaggcgg acgttctgcc atttatggag agagaaatgg cgcatgtgag ggatttgtgg
     1501 aagtcaagga gagccggtgc cgccaccccc gcagccagct cagtggaagt ggagtctcag
     1561 cttcagcggc tgtcttctgc tgtggagaac tcaaagactg ttggcaccga gctcgctgct
     1621 ctggtggagc ctttggctga tgcagctatc ggatgggtag ctagcactct gacaaccaca
     1681 cgtgaagcga tagtagcaaa cgatgaaagc accgaagcca agcgcagcga gctgaaacag
     1741 cttctgaaaa tgcagtctcg tctcagaaaa gtgagaccaa tacttgagaa acatcgcaaa
     1801 gtttatcagt ccgttctcca aaggggccag ctgcccaatg cccaagcaac cctctttgct
     1861 cttcaactta tagccaagca acgattcggg gacgacgttt ctggccttgt ggaagcggta
     1921 cctctcctct ccacgactgc gcccctgacc ggcggcgcgc ccggcagcct gcaccacatc
     1981 ctggacgccc agaagatgct gtggaaccac agggacagga acctgcctcc tctggcccct
     2041 ctgggccccc accaccatca ccaccattga gtctagacac gtgattaatt aaggatcccc
     2101 cgacctcgac ctctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt
     2161 ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttggtc gagatccctc
     2221 ggagatctct agctagagga tcgatccccg ccccggacga actaaacctg actacgacat
     2281 ctctgcccct tcttcgcggg gcagtgcatg taatcccttc agttggttgg tacaacttgc
     2341 caactgggcc ctgttccaca tgtgacacgg ggggggacca aacacaaagg ggttctctga
     2401 ctgtagttga catccttata aatggatgtg cacatttgcc aacactgagt ggctttcatc
     2461 ctggagcaga ctttgcagtc tgtggactgc aacacaacat tgcctttatg tgtaactctt
     2521 ggctgaagct cttacaccaa tgctggggga catgtacctc ccaggggccc aggaagacta
     2581 cgggaggcta caccaacgtc aatcagaggg gcctgtgtag ctaccgataa gcggaccctc
     2641 aagagggcat tagcaatagt gtttataagg cccccttgtt aaccctaaac gggtagcata
     2701 tgcttcccgg gtagtagtat atactatcca gactaaccct aattcaatag catatgttac
     2761 ccaacgggaa gcatatgcta tcgaattagg gttagtaaaa gggtcctaag gaacagcgat
     2821 atctcccacc ccatgagctg tcacggtttt atttacatgg ggtcaggatt ccacgagggt
     2881 agtgaaccat tttagtcaca agggcagtgg ctgaagatca aggagcgggc agtgaactct
     2941 cctgaatctt cgcctgcttc ttcattctcc ttcgtttagc taatagaata actgctgagt
     3001 tgtgaacagt aaggtgtatg tgaggtgctc gaaaacaagg tttcaggtga cgcccccaga
     3061 ataaaatttg gacggggggt tcagtggtgg cattgtgcta tgacaccaat ataaccctca
     3121 caaacccctt gggcaataaa tactagtgta ggaatgaaac attctgaata tctttaacaa
     3181 tagaaatcca tggggtgggg acaagccgta aagactggat gtccatctca cacgaattta
     3241 tggctatggg caacacataa tcctagtgca atatgatact ggggttatta agatgtgtcc
     3301 caggcaggga ccaagacagg tgaaccatgt tgttacactc tatttgtaac aaggggaaag
     3361 agagtggacg ccgacagcag cggactccac tggttgtctc taacaccccc gaaaattaaa
     3421 cggggctcca cgccaatggg gcccataaac aaagacaagt ggccactctt ttttttgaaa
     3481 ttgtggagtg ggggcacgcg tcagccccca cacgccgccc tgcggttttg gactgtaaaa
     3541 taagggtgta ataacttggc tgattgtaac cccgctaacc actgcggtca aaccacttgc
     3601 ccacaaaacc actaatggca ccccggggaa tacctgcata agtaggtggg cgggccaaga
     3661 taggggcgcg attgctgcga tctggaggac aaattacaca cacttgcgcc tgagcgccaa
     3721 gcacagggtt gttggtcctc atattcacga ggtcgctgag agcacggtgg gctaatgttg
     3781 ccatgggtag catatactac ccaaatatct ggatagcata tgctatccta atctatatct
     3841 gggtagcata ggctatccta atctatatct gggtagcata tgctatccta atctatatct
     3901 gggtagtata tgctatccta atttatatct gggtagcata ggctatccta atctatatct
     3961 gggtagcata tgctatccta atctatatct gggtagtata tgctatccta atctgtatcc
     4021 gggtagcata tgctatccta atagagatta gggtagtata tgctatccta atttatatct
     4081 gggtagcata tactacccaa atatctggat agcatatgct atcctaatct atatctgggt
     4141 agcatatgct atcctaatct atatctgggt agcataggct atcctaatct atatctgggt
     4201 agcatatgct atcctaatct atatctgggt agtatatgct atcctaattt atatctgggt
     4261 agcataggct atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt
     4321 agtatatgct atcctaatct gtatccgggt agcatatgct atcctcatga taagctgtca
     4381 aacatgagaa ttttcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     4441 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     4501 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4561 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     4621 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     4681 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     4741 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     4801 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga
     4861 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     4921 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     4981 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     5041 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     5101 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac
     5161 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     5221 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     5281 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     5341 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     5401 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     5461 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     5521 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     5581 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     5641 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     5701 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     5761 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     5821 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     5881 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5941 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     6001 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     6061 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     6121 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     6181 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     6241 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     6301 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     6361 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     6421 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     6481 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
     6541 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     6601 tcacacagga aacagctatg accatgatta cgccaagctc tagctagagg tcgaccaatt
     6661 ctcatgtttg acagcttatc atcgcagatc cgggcaacgt tgttgccatt gctgcaggcg
     6721 cagaactggt aggtatggaa gatctataca ttgaatcaat attggcaatt agccatatta
     6781 gtcattggtt atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta
     6841 tatcataata tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga
     6901 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     6961 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     7021 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     7081 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     7141 catatgccaa gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     7201 gcccagtaca tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc
     7261 gctattacca tg
//
