LOCUS       sequence-274735-        7518 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Leishmania infantum ectodomain in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7518)
  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 7518)
  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: D82_Li_04_0170
FEATURES             Location/Qualifiers
     source          1..7518
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     1086..1105
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1128..1137
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2676..2693
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2745..2800
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2799..2818)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5036..5054)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5122..5226
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5227..5295,5296..6087)
                     /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(5445..5464)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6258..6846
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6747..6766
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7000..7017
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7134..7155
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7170..7175,7176..7193,7194..7200)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7208..7224
                     /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     7213..7235
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7232..7248
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7232..7248
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agatcctcac tctcttccgc atcgctgtct gcgagggcca gctgttgggc tcgcggttga
      661 ggacaaactc ttcgcggtct ttccagtact cttggatcgg aaacccgtcg gcctccgaac
      721 ggtactccgc caccgaggga cctgagcgag tccgcatcga ccggatcgga aaacctctcg
      781 agaaaggcgt ctaaccagtc acagtcgcaa ggtaggctga gcaccgtggc gggcggcagc
      841 gggtggcggt cggggttgtt tctggcggag gtgctgctga tgatgtaatt aaagtaggcg
      901 gtcttgagac ggcggatggt cgaggtgagg tgtggcaggc ttgagatcca gctgttgggg
      961 tgagtactcc ctctcaaaag cgggcattac ttctgcgcta agattgtcag tttccaaaaa
     1021 cgaggaggat ttgatattca cctggcccga tctggccata cacttgagtg acaatgacat
     1081 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caagtttgcc accatggagt
     1141 ttcagaccca ggtactcatg tccctgctgc tctgcatgtc tggtgcggcc gcctacacaa
     1201 cgagtcaatg tcgggaagtc agcagatgtg acgtagacgg caccctggtg tacgcgttct
     1261 ccggggagtg tgtctgctgc cccgagatgc ccacaggaga catcctgcca gtacaatgtg
     1321 tcccatcaac agctcactgc ccgtcagtac ccagctgtcg cgcatgtgat acaaccgcgg
     1381 gcatgtgtga cgcctgtatg acaggctatg tgctggaacc aacaaatatt tgtaacgaat
     1441 gtctggcttc catgtgcgac gtctgccagt ctagcagcga aggtaagcga tgtaagctgt
     1501 gctccgttgg gtactattgg gtttctggcg gtgagtgtca cgcagtgagc aactcccaag
     1561 ctagcgagtc taatatgcag tgtcgcgtgg ccttttgcag cacctgtgcg agcaccactg
     1621 catccacctg cgcaaaatgc aacgacgggt acgctaagga ctccaacgga ttttgcgttt
     1681 ccgactgtaa tgtccaacag tgtgacatgt gttataccgg gaatagtgcc gactgtcagc
     1741 tctgctccgc cggcaacacc tggtctgcca cgggatgttc cgctggagcc tcaggatgca
     1801 gggtgggaca ctgcacaact tgcgtagcag gcgacaacac aaattgcgcg acatgcgagt
     1861 ccggatacac actcaccgct gggtactgcc tgccacagca actttgtgca gatgcgaatt
     1921 gcgccagctg cccatccgac gcgggtactt gtacacaatg cgccaatggc tatggtcttg
     1981 tggacggagc atgcgttcga tgtcaggttc ctaactgttt ctcctgcgat tctgatgcca
     2041 ataaatgcgc ccagtgcgca cccaactact atctgactcc actgcttact tgcagccccg
     2101 tggcttgtaa tatcgaacac tgcatgcaat gcgatcctca gactccttca cggtgccaag
     2161 aatgtgtggc accgtatgtg gtggattcct atgacggact gtgcaggctg agcgatgcct
     2221 gtagcgtccc caactgtaag aaatgtgaga cagaaacaag cagactgtgc gccgagtgcg
     2281 acaccggata cacccttacc gccgacgcaa caagctgtag ctcccctacg actcagccct
     2341 gcgacgtgga gcactgcaac acctgtgtta acggagatag cacgaggtgc gcttattgta
     2401 acaccgggta ctacgtctct gatggaaaat gcaagaccat gcaggggtgt tatgtctcca
     2461 attgtgccca gtgcatgctg ctggattcaa ctaagtgctc cacatgcatg aaaggttatc
     2521 tcttgacgag ttcctatggc tgtgtgtccc agaaggtcat caacggtgta gcagcccctg
     2581 gcgcgcccgg cagcctgcac cacatcctgg acgcccagaa gatgctgtgg aaccacaggg
     2641 acaggaacct gcctcctctg gcccctctgg gcccccacca ccatcaccac cattgagtct
     2701 agacacgtga ttaattaagg atcccccgac ctcgacctct ggctaataaa ggaaatttat
     2761 tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag
     2821 ggcaaatcat ttggtcgaga tccctcggag atctctagct agaggatcga tccccgcccc
     2881 ggacgaacta aacctgacta cgacatctct gccccttctt cgcggggcag tgcatgtaat
     2941 cccttcagtt ggttggtaca acttgccaac tgggccctgt tccacatgtg acacgggggg
     3001 ggaccaaaca caaaggggtt ctctgactgt agttgacatc cttataaatg gatgtgcaca
     3061 tttgccaaca ctgagtggct ttcatcctgg agcagacttt gcagtctgtg gactgcaaca
     3121 caacattgcc tttatgtgta actcttggct gaagctctta caccaatgct gggggacatg
     3181 tacctcccag gggcccagga agactacggg aggctacacc aacgtcaatc agaggggcct
     3241 gtgtagctac cgataagcgg accctcaaga gggcattagc aatagtgttt ataaggcccc
     3301 cttgttaacc ctaaacgggt agcatatgct tcccgggtag tagtatatac tatccagact
     3361 aaccctaatt caatagcata tgttacccaa cgggaagcat atgctatcga attagggtta
     3421 gtaaaagggt cctaaggaac agcgatatct cccaccccat gagctgtcac ggttttattt
     3481 acatggggtc aggattccac gagggtagtg aaccatttta gtcacaaggg cagtggctga
     3541 agatcaagga gcgggcagtg aactctcctg aatcttcgcc tgcttcttca ttctccttcg
     3601 tttagctaat agaataactg ctgagttgtg aacagtaagg tgtatgtgag gtgctcgaaa
     3661 acaaggtttc aggtgacgcc cccagaataa aatttggacg gggggttcag tggtggcatt
     3721 gtgctatgac accaatataa ccctcacaaa ccccttgggc aataaatact agtgtaggaa
     3781 tgaaacattc tgaatatctt taacaataga aatccatggg gtggggacaa gccgtaaaga
     3841 ctggatgtcc atctcacacg aatttatggc tatgggcaac acataatcct agtgcaatat
     3901 gatactgggg ttattaagat gtgtcccagg cagggaccaa gacaggtgaa ccatgttgtt
     3961 acactctatt tgtaacaagg ggaaagagag tggacgccga cagcagcgga ctccactggt
     4021 tgtctctaac acccccgaaa attaaacggg gctccacgcc aatggggccc ataaacaaag
     4081 acaagtggcc actctttttt ttgaaattgt ggagtggggg cacgcgtcag cccccacacg
     4141 ccgccctgcg gttttggact gtaaaataag ggtgtaataa cttggctgat tgtaaccccg
     4201 ctaaccactg cggtcaaacc acttgcccac aaaaccacta atggcacccc ggggaatacc
     4261 tgcataagta ggtgggcggg ccaagatagg ggcgcgattg ctgcgatctg gaggacaaat
     4321 tacacacact tgcgcctgag cgccaagcac agggttgttg gtcctcatat tcacgaggtc
     4381 gctgagagca cggtgggcta atgttgccat gggtagcata tactacccaa atatctggat
     4441 agcatatgct atcctaatct atatctgggt agcataggct atcctaatct atatctgggt
     4501 agcatatgct atcctaatct atatctgggt agtatatgct atcctaattt atatctgggt
     4561 agcataggct atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt
     4621 agtatatgct atcctaatct gtatccgggt agcatatgct atcctaatag agattagggt
     4681 agtatatgct atcctaattt atatctgggt agcatatact acccaaatat ctggatagca
     4741 tatgctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagca
     4801 taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagta
     4861 tatgctatcc taatttatat ctgggtagca taggctatcc taatctatat ctgggtagca
     4921 tatgctatcc taatctatat ctgggtagta tatgctatcc taatctgtat ccgggtagca
     4981 tatgctatcc tcatgataag ctgtcaaaca tgagaatttt cttgaagacg aaagggcctc
     5041 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     5101 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     5161 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     5221 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     5281 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     5341 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     5401 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     5461 ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     5521 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     5581 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     5641 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     5701 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     5761 acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     5821 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     5881 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     5941 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     6001 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     6061 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     6121 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     6181 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     6241 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     6301 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     6361 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     6421 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     6481 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     6541 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     6601 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     6661 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     6721 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     6781 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     6841 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     6901 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     6961 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     7021 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     7081 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     7141 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     7201 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     7261 aagctctagc tagaggtcga ccaattctca tgtttgacag cttatcatcg cagatccggg
     7321 caacgttgtt gccattgctg caggcgcaga actggtaggt atggaagatc tatacattga
     7381 atcaatattg gcaattagcc atattagtca ttggttatat agcataaatc aatattggct
     7441 attggccatt gcatacgttg tatctatatc ataatatgta catttatatt ggctcatgtc
     7501 caatatgacc gccatgtt
//
