LOCUS       sequence-274748-        6708 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 6708)
  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 6708)
  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: D78_Ld_362280
FEATURES             Location/Qualifiers
     source          1..6708
                     /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             1486..1503
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1555..1610
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1609..1628)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3846..3864)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3932..4036
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4037..4105,4106..4897)
                     /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(4255..4274)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5068..5656
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5557..5576
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5810..5827
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5944..5965
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5980..5985,5986..6003,6004..6010)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6018..6034
                     /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     6023..6045
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6042..6058
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6042..6058
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        6329..6708
                     /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 ttccagtaca 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 gccctgacct accactttgt agattcaaag
      841 cccctgtgct tcagtgagat catcgagaat gttcagacct ctcagatcac cggagtgtac
      901 gactggaaac cgtctgccca cagcccggcc agtcaggtgc gcctgcggct gagcgctaaa
      961 gatggcaccg gcaacgtcta ctatgacaaa gagatgatgg aaggggaaca ttcatttgca
     1021 gtgcagctga accctaacgt ccttagcggg gaacagctca tttgctttac agccagcagc
     1081 aattttgtcg ccagcgaaga gaaaccagta aaagtgagca ttgagctgga ccaggccccc
     1141 aagaatgagt ttaacgccga caaggtggtc gttgagacaa tacagaagcg gcgccagata
     1201 gacgggttgg acgtgtacac gtaccaagag gcaggtggag agctgaaaga tatcctgcaa
     1261 cccagagctt atctggagac aatcgaccgc gagttgtcag caatggaacg cttgttggac
     1321 cagttggtga ctaacctggg gacaagcgtg atgagagagt cccgtatgag agagacatcc
     1381 gagagtacgg gcgcgcccgg cagcctgcac cacatcctgg acgcccagaa gatgctgtgg
     1441 aaccacaggg acaggaacct gcctcctctg gcccctctgg gcccccacca ccatcaccac
     1501 cattgagtct agacacgtga ttaattaagg atcccccgac ctcgacctct ggctaataaa
     1561 ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga
     1621 catatgggag ggcaaatcat ttggtcgaga tccctcggag atctctagct agaggatcga
     1681 tccccgcccc ggacgaacta aacctgacta cgacatctct gccccttctt cgcggggcag
     1741 tgcatgtaat cccttcagtt ggttggtaca acttgccaac tgggccctgt tccacatgtg
     1801 acacgggggg ggaccaaaca caaaggggtt ctctgactgt agttgacatc cttataaatg
     1861 gatgtgcaca tttgccaaca ctgagtggct ttcatcctgg agcagacttt gcagtctgtg
     1921 gactgcaaca caacattgcc tttatgtgta actcttggct gaagctctta caccaatgct
     1981 gggggacatg tacctcccag gggcccagga agactacggg aggctacacc aacgtcaatc
     2041 agaggggcct gtgtagctac cgataagcgg accctcaaga gggcattagc aatagtgttt
     2101 ataaggcccc cttgttaacc ctaaacgggt agcatatgct tcccgggtag tagtatatac
     2161 tatccagact aaccctaatt caatagcata tgttacccaa cgggaagcat atgctatcga
     2221 attagggtta gtaaaagggt cctaaggaac agcgatatct cccaccccat gagctgtcac
     2281 ggttttattt acatggggtc aggattccac gagggtagtg aaccatttta gtcacaaggg
     2341 cagtggctga agatcaagga gcgggcagtg aactctcctg aatcttcgcc tgcttcttca
     2401 ttctccttcg tttagctaat agaataactg ctgagttgtg aacagtaagg tgtatgtgag
     2461 gtgctcgaaa acaaggtttc aggtgacgcc cccagaataa aatttggacg gggggttcag
     2521 tggtggcatt gtgctatgac accaatataa ccctcacaaa ccccttgggc aataaatact
     2581 agtgtaggaa tgaaacattc tgaatatctt taacaataga aatccatggg gtggggacaa
     2641 gccgtaaaga ctggatgtcc atctcacacg aatttatggc tatgggcaac acataatcct
     2701 agtgcaatat gatactgggg ttattaagat gtgtcccagg cagggaccaa gacaggtgaa
     2761 ccatgttgtt acactctatt tgtaacaagg ggaaagagag tggacgccga cagcagcgga
     2821 ctccactggt tgtctctaac acccccgaaa attaaacggg gctccacgcc aatggggccc
     2881 ataaacaaag acaagtggcc actctttttt ttgaaattgt ggagtggggg cacgcgtcag
     2941 cccccacacg ccgccctgcg gttttggact gtaaaataag ggtgtaataa cttggctgat
     3001 tgtaaccccg ctaaccactg cggtcaaacc acttgcccac aaaaccacta atggcacccc
     3061 ggggaatacc tgcataagta ggtgggcggg ccaagatagg ggcgcgattg ctgcgatctg
     3121 gaggacaaat tacacacact tgcgcctgag cgccaagcac agggttgttg gtcctcatat
     3181 tcacgaggtc gctgagagca cggtgggcta atgttgccat gggtagcata tactacccaa
     3241 atatctggat agcatatgct atcctaatct atatctgggt agcataggct atcctaatct
     3301 atatctgggt agcatatgct atcctaatct atatctgggt agtatatgct atcctaattt
     3361 atatctgggt agcataggct atcctaatct atatctgggt agcatatgct atcctaatct
     3421 atatctgggt agtatatgct atcctaatct gtatccgggt agcatatgct atcctaatag
     3481 agattagggt agtatatgct atcctaattt atatctgggt agcatatact acccaaatat
     3541 ctggatagca tatgctatcc taatctatat ctgggtagca tatgctatcc taatctatat
     3601 ctgggtagca taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat
     3661 ctgggtagta tatgctatcc taatttatat ctgggtagca taggctatcc taatctatat
     3721 ctgggtagca tatgctatcc taatctatat ctgggtagta tatgctatcc taatctgtat
     3781 ccgggtagca tatgctatcc tcatgataag ctgtcaaaca tgagaatttt cttgaagacg
     3841 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     3901 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     3961 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4021 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     4081 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     4141 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     4201 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     4261 tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     4321 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     4381 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     4441 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     4501 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     4561 gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga
     4621 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     4681 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     4741 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     4801 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     4861 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     4921 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     4981 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5041 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5101 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5161 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     5221 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5281 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5341 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     5401 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     5461 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     5521 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     5581 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     5641 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     5701 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     5761 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     5821 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     5881 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     5941 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     6001 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
     6061 tgattacgcc aagctctagc tagaggtcga ccaattctca tgtttgacag cttatcatcg
     6121 cagatccggg caacgttgtt gccattgctg caggcgcaga actggtaggt atggaagatc
     6181 tatacattga atcaatattg gcaattagcc atattagtca ttggttatat agcataaatc
     6241 aatattggct attggccatt gcatacgttg tatctatatc ataatatgta catttatatt
     6301 ggctcatgtc caatatgacc gccatgttga cattgattat tgactagtta ttaatagtaa
     6361 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     6421 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     6481 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     6541 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccacgtcc gccccctatt
     6601 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttacgggac
     6661 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatg
//
