LOCUS       sequence-274527-        6555 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 6555)
  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 6555)
  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: D12_Ld_040370
FEATURES             Location/Qualifiers
     source          1..6555
                     /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             1333..1350
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1402..1457
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1456..1475)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3693..3711)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3779..3883
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3884..3952,3953..4744)
                     /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(4102..4121)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4915..5503
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5404..5423
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5657..5674
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5791..5812
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5827..5832,5833..5850,5851..5857)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5865..5881
                     /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     5870..5892
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5889..5905
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5889..5905
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        6176..6555
                     /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 gccaccgata tgctgctggc ccccagcgac
      841 agcatcccct tcaccacctt cggccagctg cagcaggtct gcgccgccac acaggccgga
      901 agcacccctg tggccatgta caccgccacc atgaccgccc tggtggccaa cctgatgagc
      961 gagaagaaag tgacccacgc ctacgtgtcc gcctacctga acgccgaccg gcggtggact
     1021 tggtttgtgt acgagggcga cgaagtgatc gagatgctga tgaacagcag ccagtgggcc
     1081 accggcaacc ccacaggcgc ccacaccacc taccggcacg tggtgtacag caccaccgac
     1141 aagggcctga tcagcgccac aggctacgac aactaccccg tcgtgtgcaa atacgagaac
     1201 cagaccgact acagcccccc tggcaagcgg ttccctggcg cgcccggcag cctgcaccac
     1261 atcctggacg cccagaagat gctgtggaac cacagggaca ggaacctgcc tcctctggcc
     1321 cctctgggcc cccaccacca tcaccaccat tgagtctaga cacgtgatta attaaggatc
     1381 ccccgacctc gacctctggc taataaagga aatttatttt cattgcaata gtgtgttgga
     1441 attttttgtg tctctcactc ggaaggacat atgggagggc aaatcatttg gtcgagatcc
     1501 ctcggagatc tctagctaga ggatcgatcc ccgccccgga cgaactaaac ctgactacga
     1561 catctctgcc ccttcttcgc ggggcagtgc atgtaatccc ttcagttggt tggtacaact
     1621 tgccaactgg gccctgttcc acatgtgaca cgggggggga ccaaacacaa aggggttctc
     1681 tgactgtagt tgacatcctt ataaatggat gtgcacattt gccaacactg agtggctttc
     1741 atcctggagc agactttgca gtctgtggac tgcaacacaa cattgccttt atgtgtaact
     1801 cttggctgaa gctcttacac caatgctggg ggacatgtac ctcccagggg cccaggaaga
     1861 ctacgggagg ctacaccaac gtcaatcaga ggggcctgtg tagctaccga taagcggacc
     1921 ctcaagaggg cattagcaat agtgtttata aggccccctt gttaacccta aacgggtagc
     1981 atatgcttcc cgggtagtag tatatactat ccagactaac cctaattcaa tagcatatgt
     2041 tacccaacgg gaagcatatg ctatcgaatt agggttagta aaagggtcct aaggaacagc
     2101 gatatctccc accccatgag ctgtcacggt tttatttaca tggggtcagg attccacgag
     2161 ggtagtgaac cattttagtc acaagggcag tggctgaaga tcaaggagcg ggcagtgaac
     2221 tctcctgaat cttcgcctgc ttcttcattc tccttcgttt agctaataga ataactgctg
     2281 agttgtgaac agtaaggtgt atgtgaggtg ctcgaaaaca aggtttcagg tgacgccccc
     2341 agaataaaat ttggacgggg ggttcagtgg tggcattgtg ctatgacacc aatataaccc
     2401 tcacaaaccc cttgggcaat aaatactagt gtaggaatga aacattctga atatctttaa
     2461 caatagaaat ccatggggtg gggacaagcc gtaaagactg gatgtccatc tcacacgaat
     2521 ttatggctat gggcaacaca taatcctagt gcaatatgat actggggtta ttaagatgtg
     2581 tcccaggcag ggaccaagac aggtgaacca tgttgttaca ctctatttgt aacaagggga
     2641 aagagagtgg acgccgacag cagcggactc cactggttgt ctctaacacc cccgaaaatt
     2701 aaacggggct ccacgccaat ggggcccata aacaaagaca agtggccact cttttttttg
     2761 aaattgtgga gtgggggcac gcgtcagccc ccacacgccg ccctgcggtt ttggactgta
     2821 aaataagggt gtaataactt ggctgattgt aaccccgcta accactgcgg tcaaaccact
     2881 tgcccacaaa accactaatg gcaccccggg gaatacctgc ataagtaggt gggcgggcca
     2941 agataggggc gcgattgctg cgatctggag gacaaattac acacacttgc gcctgagcgc
     3001 caagcacagg gttgttggtc ctcatattca cgaggtcgct gagagcacgg tgggctaatg
     3061 ttgccatggg tagcatatac tacccaaata tctggatagc atatgctatc ctaatctata
     3121 tctgggtagc ataggctatc ctaatctata tctgggtagc atatgctatc ctaatctata
     3181 tctgggtagt atatgctatc ctaatttata tctgggtagc ataggctatc ctaatctata
     3241 tctgggtagc atatgctatc ctaatctata tctgggtagt atatgctatc ctaatctgta
     3301 tccgggtagc atatgctatc ctaatagaga ttagggtagt atatgctatc ctaatttata
     3361 tctgggtagc atatactacc caaatatctg gatagcatat gctatcctaa tctatatctg
     3421 ggtagcatat gctatcctaa tctatatctg ggtagcatag gctatcctaa tctatatctg
     3481 ggtagcatat gctatcctaa tctatatctg ggtagtatat gctatcctaa tttatatctg
     3541 ggtagcatag gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg
     3601 ggtagtatat gctatcctaa tctgtatccg ggtagcatat gctatcctca tgataagctg
     3661 tcaaacatga gaattttctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt
     3721 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     3781 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     3841 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     3901 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     3961 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     4021 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     4081 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
     4141 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     4201 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     4261 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     4321 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     4381 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac
     4441 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     4501 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     4561 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     4621 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     4681 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     4741 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     4801 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     4861 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     4921 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     4981 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5041 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     5101 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5161 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5221 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5281 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     5341 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     5401 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     5461 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     5521 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     5581 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     5641 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     5701 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     5761 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
     5821 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
     5881 atttcacaca ggaaacagct atgaccatga ttacgccaag ctctagctag aggtcgacca
     5941 attctcatgt ttgacagctt atcatcgcag atccgggcaa cgttgttgcc attgctgcag
     6001 gcgcagaact ggtaggtatg gaagatctat acattgaatc aatattggca attagccata
     6061 ttagtcattg gttatatagc ataaatcaat attggctatt ggccattgca tacgttgtat
     6121 ctatatcata atatgtacat ttatattggc tcatgtccaa tatgaccgcc atgttgacat
     6181 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
     6241 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     6301 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     6361 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     6421 tatcatatgc caagtccgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     6481 tatgcccagt acatgacctt acgggacttt cctacttggc agtacatcta cgtattagtc
     6541 atcgctatta ccatg
//
