LOCUS       sequence-274510-        7800 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 7800)
  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 7800)
  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: D3_Ld_030930
FEATURES             Location/Qualifiers
     source          1..7800
                     /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             2578..2595
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2647..2702
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2701..2720)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4938..4956)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5024..5128
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5129..5197,5198..5989)
                     /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(5347..5366)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6160..6748
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6649..6668
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6902..6919
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7036..7057
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7072..7077,7078..7095,7096..7102)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7110..7126
                     /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     7115..7137
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7134..7150
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7134..7150
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7421..7800
                     /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 gccaccgctg atgctgtgtc cagccccgtg
      841 atcctgccca gaggccggtc caacaccaaa gtgaccctgt acgtgcggac cggcagagtg
      901 tccatccccc tggaatggat cgacggcaaa gaggtgttct tcgagtacac cggcagactg
      961 tacgaagtgg gcaacagcgg ccccatgctg cctggcccca ccctgaaagt gaaccctggc
     1021 ggcagaatcg tgctgaagct ggtcaacgac ctgggcaaag agggcatggc caacatgacc
     1081 ggccggatga acaccctgca cggccccaac atcaccaacg tgcacttcca cggcatgcac
     1141 agcgacccca agatggacaa ccccttcaag gtggccctgc ccggcgagac actggtgtac
     1201 acaatcaacg tgccccggga ccacgagccc ggcctgcact ggtatcacgc ccacagccac
     1261 ggggccgtgt actaccacgt gatgggcggc atgttcggcg ccatcgacgt gggcgagggc
     1321 gacttcacca agacccccag acaccccttc cacggctggg acagccagat cctgatggtg
     1381 cacctgtacc ggctgaacac cagcgagaga tgcgacggca tgcccatgag cagcctggac
     1441 gccgccatgt ccagcctgct gcccttcgac ccccggatcg tggaccggaa gggcaacgag
     1501 tacgagatgc ccgccgagct gttcctggtc aacggccagc accggcccac cgtcagagtg
     1561 aagaggggcc accctatgct gctgcggatc ggctttgccg ccggaagctg ctacatcaac
     1621 atgagcctgc ccaagcagtg cgccttccac atgacagcca tcgacggcgc ccagctgccc
     1681 cggaccgtgg aagtggtgga aggctggctg tacttcacca ccgccacccg gcgaagcctg
     1741 gtggtcgtgt gtgacgaaga gggcacctac cccgtgtccc acaccggcga tcccagcgac
     1801 gccatcttct acctgaagtc ccggggcaag caccagaaag gcggcagagc cgtggccttc
     1861 cccgtgacca tgcacaagta cagccccggc tacctgcagc tgaagggcca gaacaccttc
     1921 tacagagaga tcagcttcag ccagcgggac ctgcccatca ccaagcccta ctacgtgctg
     1981 ggccagggcc ccaactgcca gagcctgcag aacagcagca cctgtcacta cgagtacttc
     2041 cagggcgaga tcggcagacg gatcgagggc taccacggct tcaccgtgcc tctgcacgcc
     2101 gtggtcaccg ccagagtgta cggcgacccc accgacaaga ggccccaccc cctgcacttt
     2161 cacgtgaacc acttccagtt tctgagcttc gagcccagac ctggcggcaa ccacgagaac
     2221 gagacaatgg ctatgtacgg cgtgcactcc ggcgagttcc gggacaccat ccccatcctg
     2281 gacggcgtga ccaccatccg gtggaaggcc gccacctatg tgggcgaggt ggtgtaccac
     2341 tgccacgccc tgcagcacga ggaccggggc atgatgatga gctacctggt gtacagcccc
     2401 cggtccaaag agggaaaggc cgtccagaga tacgagcaga gccagaccgg cgccttcctg
     2461 ctgacctggc ctcacagatc tggcgcgccc ggcagcctgc accacatcct ggacgcccag
     2521 aagatgctgt ggaaccacag ggacaggaac ctgcctcctc tggcccctct gggcccccac
     2581 caccatcacc accattgagt ctagacacgt gattaattaa ggatcccccg acctcgacct
     2641 ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct
     2701 cactcggaag gacatatggg agggcaaatc atttggtcga gatccctcgg agatctctag
     2761 ctagaggatc gatccccgcc ccggacgaac taaacctgac tacgacatct ctgccccttc
     2821 ttcgcggggc agtgcatgta atcccttcag ttggttggta caacttgcca actgggccct
     2881 gttccacatg tgacacgggg ggggaccaaa cacaaagggg ttctctgact gtagttgaca
     2941 tccttataaa tggatgtgca catttgccaa cactgagtgg ctttcatcct ggagcagact
     3001 ttgcagtctg tggactgcaa cacaacattg cctttatgtg taactcttgg ctgaagctct
     3061 tacaccaatg ctgggggaca tgtacctccc aggggcccag gaagactacg ggaggctaca
     3121 ccaacgtcaa tcagaggggc ctgtgtagct accgataagc ggaccctcaa gagggcatta
     3181 gcaatagtgt ttataaggcc cccttgttaa ccctaaacgg gtagcatatg cttcccgggt
     3241 agtagtatat actatccaga ctaaccctaa ttcaatagca tatgttaccc aacgggaagc
     3301 atatgctatc gaattagggt tagtaaaagg gtcctaagga acagcgatat ctcccacccc
     3361 atgagctgtc acggttttat ttacatgggg tcaggattcc acgagggtag tgaaccattt
     3421 tagtcacaag ggcagtggct gaagatcaag gagcgggcag tgaactctcc tgaatcttcg
     3481 cctgcttctt cattctcctt cgtttagcta atagaataac tgctgagttg tgaacagtaa
     3541 ggtgtatgtg aggtgctcga aaacaaggtt tcaggtgacg cccccagaat aaaatttgga
     3601 cggggggttc agtggtggca ttgtgctatg acaccaatat aaccctcaca aaccccttgg
     3661 gcaataaata ctagtgtagg aatgaaacat tctgaatatc tttaacaata gaaatccatg
     3721 gggtggggac aagccgtaaa gactggatgt ccatctcaca cgaatttatg gctatgggca
     3781 acacataatc ctagtgcaat atgatactgg ggttattaag atgtgtccca ggcagggacc
     3841 aagacaggtg aaccatgttg ttacactcta tttgtaacaa ggggaaagag agtggacgcc
     3901 gacagcagcg gactccactg gttgtctcta acacccccga aaattaaacg gggctccacg
     3961 ccaatggggc ccataaacaa agacaagtgg ccactctttt ttttgaaatt gtggagtggg
     4021 ggcacgcgtc agcccccaca cgccgccctg cggttttgga ctgtaaaata agggtgtaat
     4081 aacttggctg attgtaaccc cgctaaccac tgcggtcaaa ccacttgccc acaaaaccac
     4141 taatggcacc ccggggaata cctgcataag taggtgggcg ggccaagata ggggcgcgat
     4201 tgctgcgatc tggaggacaa attacacaca cttgcgcctg agcgccaagc acagggttgt
     4261 tggtcctcat attcacgagg tcgctgagag cacggtgggc taatgttgcc atgggtagca
     4321 tatactaccc aaatatctgg atagcatatg ctatcctaat ctatatctgg gtagcatagg
     4381 ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagtatatg
     4441 ctatcctaat ttatatctgg gtagcatagg ctatcctaat ctatatctgg gtagcatatg
     4501 ctatcctaat ctatatctgg gtagtatatg ctatcctaat ctgtatccgg gtagcatatg
     4561 ctatcctaat agagattagg gtagtatatg ctatcctaat ttatatctgg gtagcatata
     4621 ctacccaaat atctggatag catatgctat cctaatctat atctgggtag catatgctat
     4681 cctaatctat atctgggtag cataggctat cctaatctat atctgggtag catatgctat
     4741 cctaatctat atctgggtag tatatgctat cctaatttat atctgggtag cataggctat
     4801 cctaatctat atctgggtag catatgctat cctaatctat atctgggtag tatatgctat
     4861 cctaatctgt atccgggtag catatgctat cctcatgata agctgtcaaa catgagaatt
     4921 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     4981 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     5041 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     5101 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     5161 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     5221 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     5281 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     5341 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
     5401 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     5461 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     5521 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     5581 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     5641 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact
     5701 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     5761 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     5821 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     5881 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     5941 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     6001 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     6061 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     6121 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     6181 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     6241 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     6301 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     6361 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     6421 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     6481 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     6541 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     6601 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     6661 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     6721 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     6781 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     6841 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     6901 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     6961 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     7021 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     7081 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     7141 cagctatgac catgattacg ccaagctcta gctagaggtc gaccaattct catgtttgac
     7201 agcttatcat cgcagatccg ggcaacgttg ttgccattgc tgcaggcgca gaactggtag
     7261 gtatggaaga tctatacatt gaatcaatat tggcaattag ccatattagt cattggttat
     7321 atagcataaa tcaatattgg ctattggcca ttgcatacgt tgtatctata tcataatatg
     7381 tacatttata ttggctcatg tccaatatga ccgccatgtt gacattgatt attgactagt
     7441 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     7501 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     7561 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     7621 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     7681 ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     7741 accttacggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
//
