LOCUS       sequence-262351-        7266 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 7266)
  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 7266)
  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: D59_Ld_312380
FEATURES             Location/Qualifiers
     source          1..7266
                     /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"
     regulatory      1194..1203
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2430..2447
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2499..2554
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2553..2572)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4790..4808)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4876..4980
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4981..5049,5050..5841)
                     /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(5199..5218)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6012..6600
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6501..6520
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6754..6771
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6888..6909
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6924..6929,6930..6947,6948..6954)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6962..6978
                     /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     6967..6989
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6986..7002
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6986..7002
                     /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 tggccagagc cagatttctg cagctgctgc tggtcaccct gaccctgctg tctacagccg
     1261 ccctgcctgt gtccgcctgg tggtccaagg gccacatgag cgtggccctg atcgccaaga
     1321 gacacatggg cgccagcctg gtggaaaagg ccgagctggc cgccaaggtg ctgagcttca
     1381 gcggccccta ccccaagagc cccgacatgg tgcagaccgc cccctgggcc gacgacatca
     1441 agaccatcgg cctgaaaacc ctgagcacct ggcactacat caccaccccc tactacaccg
     1501 acgaggactt caccctggac gtgtcccccg tgcagaccgt gaacgtggcc agcgtgatcc
     1561 ccatgctgca gacagccatc gagaagccca ccgccaacag cgacgtgatc gtgcagagcc
     1621 tggccctgct gctgcacttc atgggcgaca tccaccagcc cctgcacaac gtgaacctgt
     1681 tcagcaacca gtaccccgag agcgacctgg gcggcaacaa gcagctggtg gtcatcgaca
     1741 gcaagggcac caagatgctg ctgcatgcct actgggacag catggccgag ggcaagagcg
     1801 gcgaggacgt gcccagacct ctgagcgagg ccgactacga cgacctgaac aacttcgccg
     1861 actacctgga agccacctac gccagcaccc tgaccgacaa agaaaagaac ctggtggaca
     1921 ccaccgagat cagcaaagag acattcgacc tggccctgaa gtacgcctac cctggcgccg
     1981 acaatggcgc caccctgagc gacgagtaca agaccaacgc caagaagatc agcgagagac
     2041 aggtgctgct ggccggctac cggctggcca aaatgctgaa caccaccctg aagtccgtgt
     2101 ccatggatac catcctgcag ggcctgaagt ccatccagag cgaggtggac accgagaaca
     2161 aggccgaggt gcacaaccac tacgaccaga agggcatctc tgccgccgtg accgccattg
     2221 tggccgtggc cctgtttatc gccggcatca tcattgccac cctggtggtg ctggctctga
     2281 agtgctacct gcccaagcgg gacagattcg gcagctacga gcacgtggcc ctgggcgcgc
     2341 ccggcagcct gcaccacatc ctggacgccc agaagatgct gtggaaccac agggacagga
     2401 acctgcctcc tctggcccct ctgggccccc accaccatca ccaccattga gtctagacac
     2461 gtgattaatt aaggatcccc cgacctcgac ctctggctaa taaaggaaat ttattttcat
     2521 tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa
     2581 tcatttggtc gagatccctc ggagatctct agctagagga tcgatccccg ccccggacga
     2641 actaaacctg actacgacat ctctgcccct tcttcgcggg gcagtgcatg taatcccttc
     2701 agttggttgg tacaacttgc caactgggcc ctgttccaca tgtgacacgg ggggggacca
     2761 aacacaaagg ggttctctga ctgtagttga catccttata aatggatgtg cacatttgcc
     2821 aacactgagt ggctttcatc ctggagcaga ctttgcagtc tgtggactgc aacacaacat
     2881 tgcctttatg tgtaactctt ggctgaagct cttacaccaa tgctggggga catgtacctc
     2941 ccaggggccc aggaagacta cgggaggcta caccaacgtc aatcagaggg gcctgtgtag
     3001 ctaccgataa gcggaccctc aagagggcat tagcaatagt gtttataagg cccccttgtt
     3061 aaccctaaac gggtagcata tgcttcccgg gtagtagtat atactatcca gactaaccct
     3121 aattcaatag catatgttac ccaacgggaa gcatatgcta tcgaattagg gttagtaaaa
     3181 gggtcctaag gaacagcgat atctcccacc ccatgagctg tcacggtttt atttacatgg
     3241 ggtcaggatt ccacgagggt agtgaaccat tttagtcaca agggcagtgg ctgaagatca
     3301 aggagcgggc agtgaactct cctgaatctt cgcctgcttc ttcattctcc ttcgtttagc
     3361 taatagaata actgctgagt tgtgaacagt aaggtgtatg tgaggtgctc gaaaacaagg
     3421 tttcaggtga cgcccccaga ataaaatttg gacggggggt tcagtggtgg cattgtgcta
     3481 tgacaccaat ataaccctca caaacccctt gggcaataaa tactagtgta ggaatgaaac
     3541 attctgaata tctttaacaa tagaaatcca tggggtgggg acaagccgta aagactggat
     3601 gtccatctca cacgaattta tggctatggg caacacataa tcctagtgca atatgatact
     3661 ggggttatta agatgtgtcc caggcaggga ccaagacagg tgaaccatgt tgttacactc
     3721 tatttgtaac aaggggaaag agagtggacg ccgacagcag cggactccac tggttgtctc
     3781 taacaccccc gaaaattaaa cggggctcca cgccaatggg gcccataaac aaagacaagt
     3841 ggccactctt ttttttgaaa ttgtggagtg ggggcacgcg tcagccccca cacgccgccc
     3901 tgcggttttg gactgtaaaa taagggtgta ataacttggc tgattgtaac cccgctaacc
     3961 actgcggtca aaccacttgc ccacaaaacc actaatggca ccccggggaa tacctgcata
     4021 agtaggtggg cgggccaaga taggggcgcg attgctgcga tctggaggac aaattacaca
     4081 cacttgcgcc tgagcgccaa gcacagggtt gttggtcctc atattcacga ggtcgctgag
     4141 agcacggtgg gctaatgttg ccatgggtag catatactac ccaaatatct ggatagcata
     4201 tgctatccta atctatatct gggtagcata ggctatccta atctatatct gggtagcata
     4261 tgctatccta atctatatct gggtagtata tgctatccta atttatatct gggtagcata
     4321 ggctatccta atctatatct gggtagcata tgctatccta atctatatct gggtagtata
     4381 tgctatccta atctgtatcc gggtagcata tgctatccta atagagatta gggtagtata
     4441 tgctatccta atttatatct gggtagcata tactacccaa atatctggat agcatatgct
     4501 atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt agcataggct
     4561 atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt agtatatgct
     4621 atcctaattt atatctgggt agcataggct atcctaatct atatctgggt agcatatgct
     4681 atcctaatct atatctgggt agtatatgct atcctaatct gtatccgggt agcatatgct
     4741 atcctcatga taagctgtca aacatgagaa ttttcttgaa gacgaaaggg cctcgtgata
     4801 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     4861 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     4921 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     4981 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     5041 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     5101 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     5161 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     5221 cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     5281 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     5341 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     5401 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     5461 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     5521 cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     5581 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     5641 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     5701 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     5761 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     5821 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     5881 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     5941 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     6001 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     6061 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     6121 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     6181 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     6241 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     6301 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     6361 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     6421 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     6481 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     6541 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     6601 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     6661 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     6721 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     6781 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     6841 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     6901 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     6961 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagctc
     7021 tagctagagg tcgaccaatt ctcatgtttg acagcttatc atcgcagatc cgggcaacgt
     7081 tgttgccatt gctgcaggcg cagaactggt aggtatggaa gatctataca ttgaatcaat
     7141 attggcaatt agccatatta gtcattggtt atatagcata aatcaatatt ggctattggc
     7201 cattgcatac gttgtatcta tatcataata tgtacattta tattggctca tgtccaatat
     7261 gaccgc
//
