LOCUS       sequence-274659-        6825 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 6825)
  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 6825)
  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: D54_ Ld_061160
FEATURES             Location/Qualifiers
     source          1..6825
                     /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             1603..1620
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1672..1727
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1726..1745)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3963..3981)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4049..4153
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4154..4222,4223..5014)
                     /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(4372..4391)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5185..5773
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5674..5693
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5927..5944
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6061..6082
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6097..6102,6103..6120,6121..6127)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6135..6151
                     /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     6140..6162
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6159..6175
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6159..6175
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        6446..6825
                     /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 gccgtgacag ccgtgcagag cggcagctgc
      841 atcttcctga ccagcccccc cagctacacc ttcatcggcg gaggcttcca catgcagctg
      901 gccctggaat accccctgcc cgccaaggac gtgcggctga gcttcgacct gcccaagagc
      961 ttcttcgtgg acgaggccga ggccgagcag ctgtaccgga tggaagtgct gagcgccgac
     1021 cccatgacac agggcacagc cgccgtggtg gtggccgacg tgacaagagc ctacagcccc
     1081 ctgcggatga gcagcagata cgccttcgac atcgaggccc ccgtgttcaa agtgaactac
     1141 accaccaacc acgtggaact gaccttccag cagctgggcg gcgacggcgc tggaagcgcc
     1201 ctggatgcct atctggccga agatggcgcc caggcccagg tgcccttccg ggccagactg
     1261 gtcatcccca tccactccag atacgaggtg ctggacacca agaccccctt cagcctgctg
     1321 cggttcatca ccggcgaggg cgcctacgtg cggcggtgcc tgacaagagt ggatgtggcc
     1381 ggcagagccg acagcagatg cctgaccggc gcctacagat ccagcgccac cgtggccaag
     1441 agcgaggaaa agccccacta caagcactgc ctggacctgc ccgtgggcct gctggctgat
     1501 ctgcctggcg cgcccggcag cctgcaccac atcctggacg cccagaagat gctgtggaac
     1561 cacagggaca ggaacctgcc tcctctggcc cctctgggcc cccaccacca tcaccaccat
     1621 tgagtctaga cacgtgatta attaaggatc ccccgacctc gacctctggc taataaagga
     1681 aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat
     1741 atgggagggc aaatcatttg gtcgagatcc ctcggagatc tctagctaga ggatcgatcc
     1801 ccgccccgga cgaactaaac ctgactacga catctctgcc ccttcttcgc ggggcagtgc
     1861 atgtaatccc ttcagttggt tggtacaact tgccaactgg gccctgttcc acatgtgaca
     1921 cgggggggga ccaaacacaa aggggttctc tgactgtagt tgacatcctt ataaatggat
     1981 gtgcacattt gccaacactg agtggctttc atcctggagc agactttgca gtctgtggac
     2041 tgcaacacaa cattgccttt atgtgtaact cttggctgaa gctcttacac caatgctggg
     2101 ggacatgtac ctcccagggg cccaggaaga ctacgggagg ctacaccaac gtcaatcaga
     2161 ggggcctgtg tagctaccga taagcggacc ctcaagaggg cattagcaat agtgtttata
     2221 aggccccctt gttaacccta aacgggtagc atatgcttcc cgggtagtag tatatactat
     2281 ccagactaac cctaattcaa tagcatatgt tacccaacgg gaagcatatg ctatcgaatt
     2341 agggttagta aaagggtcct aaggaacagc gatatctccc accccatgag ctgtcacggt
     2401 tttatttaca tggggtcagg attccacgag ggtagtgaac cattttagtc acaagggcag
     2461 tggctgaaga tcaaggagcg ggcagtgaac tctcctgaat cttcgcctgc ttcttcattc
     2521 tccttcgttt agctaataga ataactgctg agttgtgaac agtaaggtgt atgtgaggtg
     2581 ctcgaaaaca aggtttcagg tgacgccccc agaataaaat ttggacgggg ggttcagtgg
     2641 tggcattgtg ctatgacacc aatataaccc tcacaaaccc cttgggcaat aaatactagt
     2701 gtaggaatga aacattctga atatctttaa caatagaaat ccatggggtg gggacaagcc
     2761 gtaaagactg gatgtccatc tcacacgaat ttatggctat gggcaacaca taatcctagt
     2821 gcaatatgat actggggtta ttaagatgtg tcccaggcag ggaccaagac aggtgaacca
     2881 tgttgttaca ctctatttgt aacaagggga aagagagtgg acgccgacag cagcggactc
     2941 cactggttgt ctctaacacc cccgaaaatt aaacggggct ccacgccaat ggggcccata
     3001 aacaaagaca agtggccact cttttttttg aaattgtgga gtgggggcac gcgtcagccc
     3061 ccacacgccg ccctgcggtt ttggactgta aaataagggt gtaataactt ggctgattgt
     3121 aaccccgcta accactgcgg tcaaaccact tgcccacaaa accactaatg gcaccccggg
     3181 gaatacctgc ataagtaggt gggcgggcca agataggggc gcgattgctg cgatctggag
     3241 gacaaattac acacacttgc gcctgagcgc caagcacagg gttgttggtc ctcatattca
     3301 cgaggtcgct gagagcacgg tgggctaatg ttgccatggg tagcatatac tacccaaata
     3361 tctggatagc atatgctatc ctaatctata tctgggtagc ataggctatc ctaatctata
     3421 tctgggtagc atatgctatc ctaatctata tctgggtagt atatgctatc ctaatttata
     3481 tctgggtagc ataggctatc ctaatctata tctgggtagc atatgctatc ctaatctata
     3541 tctgggtagt atatgctatc ctaatctgta tccgggtagc atatgctatc ctaatagaga
     3601 ttagggtagt atatgctatc ctaatttata tctgggtagc atatactacc caaatatctg
     3661 gatagcatat gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg
     3721 ggtagcatag gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg
     3781 ggtagtatat gctatcctaa tttatatctg ggtagcatag gctatcctaa tctatatctg
     3841 ggtagcatat gctatcctaa tctatatctg ggtagtatat gctatcctaa tctgtatccg
     3901 ggtagcatat gctatcctca tgataagctg tcaaacatga gaattttctt gaagacgaaa
     3961 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     4021 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     4081 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     4141 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     4201 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     4261 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     4321 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     4381 cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc
     4441 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     4501 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     4561 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     4621 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     4681 tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     4741 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     4801 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     4861 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     4921 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     4981 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     5041 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     5101 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     5161 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     5221 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     5281 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     5341 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     5401 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     5461 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     5521 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     5581 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     5641 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     5701 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     5761 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     5821 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     5881 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     5941 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     6001 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     6061 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     6121 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
     6181 ttacgccaag ctctagctag aggtcgacca attctcatgt ttgacagctt atcatcgcag
     6241 atccgggcaa cgttgttgcc attgctgcag gcgcagaact ggtaggtatg gaagatctat
     6301 acattgaatc aatattggca attagccata ttagtcattg gttatatagc ataaatcaat
     6361 attggctatt ggccattgca tacgttgtat ctatatcata atatgtacat ttatattggc
     6421 tcatgtccaa tatgaccgcc atgttgacat tgattattga ctagttatta atagtaatca
     6481 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
     6541 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
     6601 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
     6661 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtccgcc ccctattgac
     6721 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt acgggacttt
     6781 cctacttggc agtacatcta cgtattagtc atcgctatta ccatg
//
