LOCUS       sequence-262313-        9324 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 9324)
  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 9324)
  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: D18_Ld_100590
FEATURES             Location/Qualifiers
     source          1..9324
                     /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"
     CDS             4488..4505
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4557..4612
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4611..4630)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6848..6866)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6934..7038
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7039..7107,7108..7899)
                     /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(7257..7276)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8070..8658
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8559..8578
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8812..8829
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8946..8967
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8982..8987,8988..9005,9006..9012)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9020..9036
                     /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     9025..9047
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9044..9060
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9044..9060
                     /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 ccgctgaggg aaagctgcag cccccctaca ccaagcccgt gatccagacc agacacgacg
     1261 acgccgtgca ccccgagagc ggcgcctatg ctagcgtgct gcccgtgtcc ctgaacgcta
     1321 ccgtggccga ccctttcctg acccctgcca ccctgatgga aggcgtgcgg agcaccggca
     1381 aggccatcac cccccagcac atcttcacca acgccgccct gcggctggtg ttcgacgtgc
     1441 ccaaccccgg cctggacttc gtgatcaacc tgaccagcgc ccagtacccc ctgctgcagg
     1501 accccaatgg caccctgtgg cgctctgtgc tgcctagcaa gatcgaggtg gtcaagcaga
     1561 gcagcaagta cacctacgac gtgggcaacc tgaacctgcg gagctaccag agccagagcg
     1621 agtccgtgga agtggtggct ggcgcccctg ccgccgatgc cgagtgtttc accctgaagg
     1681 cctgctgcgc cgtgaacagc agctactacc agaaatacga gctggtggac agccccgtgc
     1741 ccccttacgt gtgtaccggc gtgcccatcc cttacctgct gagacacttc accttcggcg
     1801 tggactccac cgtgaagttc agagaggccg ccgagggcgc cagctgcgag atgaaggcca
     1861 accggtggac cgaggaaagc gccacctgta ccgccaatct gaatgccgtg ctggccggcg
     1921 tgaccgccag ccctcctcac agactgccca gcagagcctt cagcctgtgc cggcagggcg
     1981 acaatctgac caactccgag tccggcagca gccccatccc cgtcgtgatt accgccttct
     2041 gcagcgagag ccggtgcatc ggctacaccc ccgccaacta cagaagcgac agaagcacct
     2101 gtctgaacta ccctctgggc ttctacacaa gcgccgacga gaacaacacc aaccagttca
     2161 gcagcttcga ggacaatgcc gccgctttct ggccctacct gttcaacgac tgcaagtgga
     2221 tcgacgacgt gaccatgtcc agccccacca ccatccagga catcagcctg cagtgtgatg
     2281 tgcccgcctc caccggctgg tccctgagct acgagaactt ctccgcccac ctgaccgaca
     2341 caatcgcccc cctgttcccc atcggcagac ccacaagagt ggccggccag tacgcctccc
     2401 ccaccgacaa ggacaccatc ctgttcgagc tggaatacgt gtccttcagc tccttcaagg
     2461 ccggcagatg gaccttccgg atctgcacca cccggcagtg cgccaccagc acagtgcctg
     2521 tggcaggcga caccggccac taccccaacg tgatgagagt gaatgtgtcc gccagcaagc
     2581 tgggcctgtc tgcctctcca cacgagcccg ccgtgggcgt cgaggtgtac agcgccacaa
     2641 cccctaccac ctcccccttt cggaccctgc tggcccccag cgtggtgtac gtggacaagg
     2701 ccgctacccc cctgagccag gacacagtga acatgaccct ggcccgggac cctcccgaga
     2761 gagtgcctag cccttacgac gccagcacca gcggcagaac caccagccgg gtgctgtgcg
     2821 agggcgacta ccggttcagc accgtgacca acaactgcca gcccctgaca gacgaggact
     2881 gtgccaccaa gtaccggggc aggcggagca agttcgaccc cgtggccaga gcctgcgcct
     2941 accctgtgcc tcccctgaat ggccccctgg tgtttaagcc cctggccgag cccaagcccc
     3001 ccagaaccta ctcccccgag gaactgcggg ccatcctgaa aacagtgaag ctgccccagt
     3061 tcctgcggac catggaaaag tcctacgagg attacgagag acagatggcc agatgggagg
     3121 gcagaagcgt gcggttccct ccaggcagag ccgccgcagc cgaggataca gatgtggtgg
     3181 ccgtggctgc cgctgatgag tccgccgtga cctacatcaa cgagagcccc atgcctggcg
     3241 ccagaggcct gagcgtgacc tgtatcgccg ccacatgcgt gctgtggacc gtggcactgc
     3301 tgcgggacgt gctggaaaag ctgttcggcc tgggcccttg gcacgaccag cggagatgcg
     3361 ccttctttgc cggcgctgtg aactgctggc actggctgag cggagcttgc gccgaaggcc
     3421 acaacgaggc cagacctcgg cagcggaggg ccacctctaa taccaccgcc acccccacaa
     3481 ccgcccagcc tagcgcccct agatccgcca cacagagcgg ccagtctggc cagcacgcca
     3541 accctgccct gcagaagaga aagggcaagg gcgcaacctt caccgccgac acccctctgc
     3601 agagcaccag aaagagcccc cacggcgctc cccgggaacc tgtgacctct gccccatttc
     3661 agcccaggga ccccacagcc acaccccaga gagagaagca ggacggcggc cagaagcggc
     3721 atctgcctag aacccacaac agccgggacg ccgagcgcga gcacagacag tctgtgtctg
     3781 ccggcaccac cggccctgcc agctcttggg gagatgcccg gcactacgtc agcgccgctc
     3841 acccacccgc ctacgatgtg ttcggcttca gcgaggcccc tcacgcccct tttggctctg
     3901 gcgccggatt cgccgctccc ggcgatatgc cttttctggg cggcgactat accggccaga
     3961 gagtgtggct gtccgacgag ggcgtgggcc tgggatacgt gccagccgcc tttgccagac
     4021 cccactccac cagctcccac cggtgccaca gctgccctag ctccccctac cctacctcca
     4081 gcccacccag ccccgagtgg cacccctttg aggacgtgta ccccaccgtg ccctccccat
     4141 gctacgcccc ctatgatgcc ccactgcccg agtttagcct gaaccccagc agcacctcct
     4201 accccggctt cgccagagct gctcctccta gccccttcgc cggctttgac gccttcccac
     4261 ctgtgtcctc cgtgcggcag tccagcaggg tggaagtcct gagcagcgat accgacggcg
     4321 cccacgggga caatagagcc gctgaagccc ctagcccagc tggcaggacc agacggccca
     4381 acagcatggt cggcgcgccc ggcagcctgc accacatcct ggacgcccag aagatgctgt
     4441 ggaaccacag ggacaggaac ctgcctcctc tggcccctct gggcccccac caccatcacc
     4501 accattgagt ctagacacgt gattaattaa ggatcccccg acctcgacct ctggctaata
     4561 aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag
     4621 gacatatggg agggcaaatc atttggtcga gatccctcgg agatctctag ctagaggatc
     4681 gatccccgcc ccggacgaac taaacctgac tacgacatct ctgccccttc ttcgcggggc
     4741 agtgcatgta atcccttcag ttggttggta caacttgcca actgggccct gttccacatg
     4801 tgacacgggg ggggaccaaa cacaaagggg ttctctgact gtagttgaca tccttataaa
     4861 tggatgtgca catttgccaa cactgagtgg ctttcatcct ggagcagact ttgcagtctg
     4921 tggactgcaa cacaacattg cctttatgtg taactcttgg ctgaagctct tacaccaatg
     4981 ctgggggaca tgtacctccc aggggcccag gaagactacg ggaggctaca ccaacgtcaa
     5041 tcagaggggc ctgtgtagct accgataagc ggaccctcaa gagggcatta gcaatagtgt
     5101 ttataaggcc cccttgttaa ccctaaacgg gtagcatatg cttcccgggt agtagtatat
     5161 actatccaga ctaaccctaa ttcaatagca tatgttaccc aacgggaagc atatgctatc
     5221 gaattagggt tagtaaaagg gtcctaagga acagcgatat ctcccacccc atgagctgtc
     5281 acggttttat ttacatgggg tcaggattcc acgagggtag tgaaccattt tagtcacaag
     5341 ggcagtggct gaagatcaag gagcgggcag tgaactctcc tgaatcttcg cctgcttctt
     5401 cattctcctt cgtttagcta atagaataac tgctgagttg tgaacagtaa ggtgtatgtg
     5461 aggtgctcga aaacaaggtt tcaggtgacg cccccagaat aaaatttgga cggggggttc
     5521 agtggtggca ttgtgctatg acaccaatat aaccctcaca aaccccttgg gcaataaata
     5581 ctagtgtagg aatgaaacat tctgaatatc tttaacaata gaaatccatg gggtggggac
     5641 aagccgtaaa gactggatgt ccatctcaca cgaatttatg gctatgggca acacataatc
     5701 ctagtgcaat atgatactgg ggttattaag atgtgtccca ggcagggacc aagacaggtg
     5761 aaccatgttg ttacactcta tttgtaacaa ggggaaagag agtggacgcc gacagcagcg
     5821 gactccactg gttgtctcta acacccccga aaattaaacg gggctccacg ccaatggggc
     5881 ccataaacaa agacaagtgg ccactctttt ttttgaaatt gtggagtggg ggcacgcgtc
     5941 agcccccaca cgccgccctg cggttttgga ctgtaaaata agggtgtaat aacttggctg
     6001 attgtaaccc cgctaaccac tgcggtcaaa ccacttgccc acaaaaccac taatggcacc
     6061 ccggggaata cctgcataag taggtgggcg ggccaagata ggggcgcgat tgctgcgatc
     6121 tggaggacaa attacacaca cttgcgcctg agcgccaagc acagggttgt tggtcctcat
     6181 attcacgagg tcgctgagag cacggtgggc taatgttgcc atgggtagca tatactaccc
     6241 aaatatctgg atagcatatg ctatcctaat ctatatctgg gtagcatagg ctatcctaat
     6301 ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat
     6361 ttatatctgg gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat
     6421 ctatatctgg gtagtatatg ctatcctaat ctgtatccgg gtagcatatg ctatcctaat
     6481 agagattagg gtagtatatg ctatcctaat ttatatctgg gtagcatata ctacccaaat
     6541 atctggatag catatgctat cctaatctat atctgggtag catatgctat cctaatctat
     6601 atctgggtag cataggctat cctaatctat atctgggtag catatgctat cctaatctat
     6661 atctgggtag tatatgctat cctaatttat atctgggtag cataggctat cctaatctat
     6721 atctgggtag catatgctat cctaatctat atctgggtag tatatgctat cctaatctgt
     6781 atccgggtag catatgctat cctcatgata agctgtcaaa catgagaatt ttcttgaaga
     6841 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     6901 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     6961 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     7021 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     7081 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     7141 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     7201 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     7261 tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac
     7321 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     7381 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     7441 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     7501 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     7561 gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc
     7621 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     7681 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     7741 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     7801 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     7861 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     7921 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     7981 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     8041 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     8101 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     8161 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     8221 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     8281 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     8341 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     8401 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     8461 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     8521 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     8581 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     8641 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     8701 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     8761 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     8821 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     8881 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     8941 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     9001 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     9061 catgattacg ccaagctcta gctagaggtc gaccaattct catgtttgac agcttatcat
     9121 cgcagatccg ggcaacgttg ttgccattgc tgcaggcgca gaactggtag gtatggaaga
     9181 tctatacatt gaatcaatat tggcaattag ccatattagt cattggttat atagcataaa
     9241 tcaatattgg ctattggcca ttgcatacgt tgtatctata tcataatatg tacatttata
     9301 ttggctcatg tccaatatga ccgc
//
