LOCUS       sequence-274726-        6582 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 6582)
  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 6582)
  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: D95_Ld_290220
FEATURES             Location/Qualifiers
     source          1..6582
                     /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             1740..1757
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1809..1864
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1863..1882)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4100..4118)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4186..4290
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4291..4359,4360..5151)
                     /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(4509..4528)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5322..5910
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5811..5830
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6064..6081
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6198..6219
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6234..6239,6240..6257,6258..6264)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6272..6288
                     /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     6277..6299
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6296..6312
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6296..6312
                     /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 gcctgggact
     1201 cttctgaaga cgctaaagcc atggccaaaa gggccaagca tgagcagatt cagttttggg
     1261 aacgtgaggt taatattttg cggcagggag aactgactcg tgcttacaac aagctgtatc
     1321 aagctgaggc cgctctcgag agtgccaggg ctaaacaagg cttcttctac actcgccccc
     1381 aggataaagc tacgatcaga ctgctcgatg aagattaccg caggactttg gtcgaggtga
     1441 aagctcttaa ggaacaagag cgacttatca tggccaaact gaagcccctc tacggggtgg
     1501 tgagtctgca tttcgcccag gaacagaagc gcacgatttc tgagtctatc aaaactgtgc
     1561 agtctcttag ctatgacaat gcttggtaca gcagcctctt ttctcttggg gaagccgaga
     1621 gcttcagcga tatcattatg ggtggcgcgc ccggcagcct gcaccacatc ctggacgccc
     1681 agaagatgct gtggaaccac agggacagga acctgcctcc tctggcccct ctgggccccc
     1741 accaccatca ccaccattga gtctagacac gtgattaatt aaggatcccc cgacctcgac
     1801 ctctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct
     1861 ctcactcgga aggacatatg ggagggcaaa tcatttggtc gagatccctc ggagatctct
     1921 agctagagga tcgatccccg ccccggacga actaaacctg actacgacat ctctgcccct
     1981 tcttcgcggg gcagtgcatg taatcccttc agttggttgg tacaacttgc caactgggcc
     2041 ctgttccaca tgtgacacgg ggggggacca aacacaaagg ggttctctga ctgtagttga
     2101 catccttata aatggatgtg cacatttgcc aacactgagt ggctttcatc ctggagcaga
     2161 ctttgcagtc tgtggactgc aacacaacat tgcctttatg tgtaactctt ggctgaagct
     2221 cttacaccaa tgctggggga catgtacctc ccaggggccc aggaagacta cgggaggcta
     2281 caccaacgtc aatcagaggg gcctgtgtag ctaccgataa gcggaccctc aagagggcat
     2341 tagcaatagt gtttataagg cccccttgtt aaccctaaac gggtagcata tgcttcccgg
     2401 gtagtagtat atactatcca gactaaccct aattcaatag catatgttac ccaacgggaa
     2461 gcatatgcta tcgaattagg gttagtaaaa gggtcctaag gaacagcgat atctcccacc
     2521 ccatgagctg tcacggtttt atttacatgg ggtcaggatt ccacgagggt agtgaaccat
     2581 tttagtcaca agggcagtgg ctgaagatca aggagcgggc agtgaactct cctgaatctt
     2641 cgcctgcttc ttcattctcc ttcgtttagc taatagaata actgctgagt tgtgaacagt
     2701 aaggtgtatg tgaggtgctc gaaaacaagg tttcaggtga cgcccccaga ataaaatttg
     2761 gacggggggt tcagtggtgg cattgtgcta tgacaccaat ataaccctca caaacccctt
     2821 gggcaataaa tactagtgta ggaatgaaac attctgaata tctttaacaa tagaaatcca
     2881 tggggtgggg acaagccgta aagactggat gtccatctca cacgaattta tggctatggg
     2941 caacacataa tcctagtgca atatgatact ggggttatta agatgtgtcc caggcaggga
     3001 ccaagacagg tgaaccatgt tgttacactc tatttgtaac aaggggaaag agagtggacg
     3061 ccgacagcag cggactccac tggttgtctc taacaccccc gaaaattaaa cggggctcca
     3121 cgccaatggg gcccataaac aaagacaagt ggccactctt ttttttgaaa ttgtggagtg
     3181 ggggcacgcg tcagccccca cacgccgccc tgcggttttg gactgtaaaa taagggtgta
     3241 ataacttggc tgattgtaac cccgctaacc actgcggtca aaccacttgc ccacaaaacc
     3301 actaatggca ccccggggaa tacctgcata agtaggtggg cgggccaaga taggggcgcg
     3361 attgctgcga tctggaggac aaattacaca cacttgcgcc tgagcgccaa gcacagggtt
     3421 gttggtcctc atattcacga ggtcgctgag agcacggtgg gctaatgttg ccatgggtag
     3481 catatactac ccaaatatct ggatagcata tgctatccta atctatatct gggtagcata
     3541 ggctatccta atctatatct gggtagcata tgctatccta atctatatct gggtagtata
     3601 tgctatccta atttatatct gggtagcata ggctatccta atctatatct gggtagcata
     3661 tgctatccta atctatatct gggtagtata tgctatccta atctgtatcc gggtagcata
     3721 tgctatccta atagagatta gggtagtata tgctatccta atttatatct gggtagcata
     3781 tactacccaa atatctggat agcatatgct atcctaatct atatctgggt agcatatgct
     3841 atcctaatct atatctgggt agcataggct atcctaatct atatctgggt agcatatgct
     3901 atcctaatct atatctgggt agtatatgct atcctaattt atatctgggt agcataggct
     3961 atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt agtatatgct
     4021 atcctaatct gtatccgggt agcatatgct atcctcatga taagctgtca aacatgagaa
     4081 ttttcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     4141 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     4201 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     4261 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     4321 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     4381 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     4441 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     4501 aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt
     4561 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     4621 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     4681 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     4741 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     4801 ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa
     4861 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     4921 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     4981 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     5041 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     5101 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     5161 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5221 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5281 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5341 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     5401 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     5461 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     5521 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     5581 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     5641 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     5701 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     5761 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     5821 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     5881 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     5941 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     6001 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     6061 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     6121 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     6181 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     6241 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     6301 aacagctatg accatgatta cgccaagctc tagctagagg tcgaccaatt ctcatgtttg
     6361 acagcttatc atcgcagatc cgggcaacgt tgttgccatt gctgcaggcg cagaactggt
     6421 aggtatggaa gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt
     6481 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata
     6541 tgtacattta tattggctca tgtccaatat gaccgccatg tt
//
