LOCUS       sequence-262341-        9492 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 9492)
  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 9492)
  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: D49_Ld_320280
FEATURES             Location/Qualifiers
     source          1..9492
                     /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             4656..4673
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4725..4780
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4779..4798)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(7016..7034)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7102..7206
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7207..7275,7276..8067)
                     /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(7425..7444)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8238..8826
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8727..8746
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8980..8997
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9114..9135
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9150..9155,9156..9173,9174..9180)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9188..9204
                     /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     9193..9215
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9212..9228
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9212..9228
                     /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 ccaccaagat cacctacgtg ggcgtgttcc aggccaccga gggcgagaca accaccgtgc
     1261 tgctgaaagt gggcgccctg aaggccgagt gtatcgccgg caccctgacc tacaacgtgg
     1321 acgacgacca cggcaacaag gccaccttcc ccaacatgca cgacccccgg atggaattca
     1381 ccgccgccat caccgacatg accagcggcg tgcggaccac ctatgtggac ctggtggtcg
     1441 tgtgcaacga caccacccct gtgggcggca ccagccgggt gtacttccgg atcagccccc
     1501 ggaccactac caccaccctg gcccccagca tgacctgcaa gtctccaggc gacgccgtcg
     1561 tcgtgggccc catgtctgcc gtgttccccg ccaacaccta caccagcagc tgccaccatc
     1621 ccagcctgga tatcgccgac gtgaccggcc ccagctacga cgccaagagc ctgttcagcg
     1681 tggaaatgac caacaagagc caggccctgg tgttcaacgc caacaaaacc gacaccctgc
     1741 ccggcaagta caccgccaca atcctggtgt cctgcggcag cagctcctgc gtggtcaagc
     1801 aggtcgtggt ggtgctgccc cccaagacca ccacaacagc cgcccctgtg atgcagtgcc
     1861 ctggcaacta caactacagc ctgatcctga acagcgacgg catcctgatc gagcagatct
     1921 tcaacctgag cgccagagcc gccgctaccg gcatgttctg cacctccggc atcatcagcg
     1981 ccaccctgaa agaacccgcc cacggggccc tgcggtggaa caacaccggc atcttcgtgt
     2041 acacccctag caccctggaa gagggcgacg atatcgacta cttcgagttc cagctgacat
     2101 gcagcaacga gggcaccaag tgcgccggaa ccgtgatcat ctccatcgtg tccgaggaaa
     2161 aggacgagat ctactacgcc atggaaggcg ccattgtgtg ccggggcaca tgcgacgctg
     2221 ccgcctggcg gacaagccct accgccctgg acctgttcga cgtgaacaag acaggcgtgg
     2281 ccgtgacccc cagaaaggac gtgcgagccg tggacggcgt ggacttcggc tttaccgcct
     2341 ccggcgacct ggtcatccac gcctacacaa ccatcggcaa cctggccgct agattcgtga
     2401 ccttctaccc tatcaccgag gccagcgtgc ccggcctgtt taacagcacc gaggtgcccg
     2461 acggcagcca cgtgtccttc gagcccagct gcctcaacaa gcaggccatc acaggccagg
     2521 gcaccgatat ctggtcctgg accgacctgg acgtgctgaa gggccacctg aacacccagg
     2581 ccgatgccta caagagcggc gacaactact accagaagtt cggcggcaac cacagacact
     2641 gcgacgtgta cagagtggac ccctgcaaat acgcccccct gatgacccct accctgaacg
     2701 acaacaacaa ctccgtgtac aacaacgcca caccctccgt gcagtggaag ctgtacatca
     2761 acgattgcga cgccacctgg gtcggaaagg cctccgtgga aagcctgcgg gccctgagac
     2821 agccagacgg ctcccccacc ttcagactgg aaggcaacgg cacctacctc gtgggggccg
     2881 tgtactccca gtccgtgaag cctgccagct ggaccagccc tctggccggc atcgtgcaca
     2941 gcgaaaccgc ctacgaagtg cggctgaaga tccacaacat gatcatcgtg gacgccctgg
     3001 ccagacccag ctccgatgcc acagcccctc tgctgcagat gagcagcgac atccagtaca
     3061 gcgccggcaa gcggaccgat acccaggaac ggctgtacag atacagcgtc ctgctgtacc
     3121 cctactttga gacaaacaag accgccgctg acatcacctc cctgagcctg aaggtggtgt
     3181 ccaccacact gctgaactcc acctggacta gccccagcaa ggtggaatgc cctatgtgca
     3241 ccggcaccat gatcagctgc accgggaccg gcaatggctt cgagacagac tgcggcgata
     3301 aggctctgat gacattcgag gctctggact acaacgcctc cgacttcgag ggcagcatcg
     3361 tgaacgatgg ctccagcggc aagagcgcct ccaccagcgt gtccccaaag aacgctctga
     3421 gagtgacctt tgccgtgcgc gtgaacggcg ctggcagcgc tgacgccaca gatgcctatc
     3481 ccgtgggcac cttcgccatc agcgtgaagc tgtcctccgg ccaggtgttc gatgtggtgg
     3541 tcaaccagac cgactacatc agcgagctga atacccggtt tctggacacc cagatctgcc
     3601 ggcccagcgc ctactggcct gtgccagatc ctctgggcag ctctgtgccc gtgaaaccct
     3661 actccgccga cacactggcc ttccctagcg gcagctacac cacagaggaa caggaagtga
     3721 agcgcgagag cgagtcctac tacggcgtgc cagagcggct gacctcccct agcgtgtgcg
     3781 tgacaccccc caacgccgac cggacccaca tcgatctgga caatatgacc gccactatcg
     3841 agaacgagaa gggacaggtc acagtgtcca tgtgcgccgt ggccgacgag cggaccttcg
     3901 gcaccaccga ctgggtcatg ttcagcttcc ccagaatcca cgaggaaaca aaagagatca
     3961 acgacgacga gattcagagc atcatcgatg acgaaaccct gagcaacaat tcctacaccg
     4021 tggcacagct gcagtacctg accctgtccg tgcacgccag cgaagtgatg tccagcctgc
     4081 tggctggcga cagcgccaac ggctatgtga acatcctgct ggacggggac aacgccccca
     4141 agtcccagcg gggcgaggaa gagacatgga tgcaccccga gtcctccagc agcagcggcc
     4201 agtctgacgc ctacctgcct tgggagatgt acgccagcag cctgaactac cggcggatcg
     4261 cccaccatac caacgctaac aagtccgaaa ccgagccctt caacttcgcc ttcatccccg
     4321 gctccctgct gcactccgga agcagcatca gagtgcccat gattatcagg gccgccatta
     4381 agtttgtgac ctacgccttc acagccgcta atagcaccca cagcagccag agagtgaaga
     4441 agtccgagag agaggaaacc ctcctgtata tcgtgcctgt gtccaggggc atctccagcg
     4501 cactgagatt cgataccgac atctacaccc cagtgatcac acagagcgga atcgcccctg
     4561 gcgcgcccgg cagcctgcac cacatcctgg acgcccagaa gatgctgtgg aaccacaggg
     4621 acaggaacct gcctcctctg gcccctctgg gcccccacca ccatcaccac cattgagtct
     4681 agacacgtga ttaattaagg atcccccgac ctcgacctct ggctaataaa ggaaatttat
     4741 tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag
     4801 ggcaaatcat ttggtcgaga tccctcggag atctctagct agaggatcga tccccgcccc
     4861 ggacgaacta aacctgacta cgacatctct gccccttctt cgcggggcag tgcatgtaat
     4921 cccttcagtt ggttggtaca acttgccaac tgggccctgt tccacatgtg acacgggggg
     4981 ggaccaaaca caaaggggtt ctctgactgt agttgacatc cttataaatg gatgtgcaca
     5041 tttgccaaca ctgagtggct ttcatcctgg agcagacttt gcagtctgtg gactgcaaca
     5101 caacattgcc tttatgtgta actcttggct gaagctctta caccaatgct gggggacatg
     5161 tacctcccag gggcccagga agactacggg aggctacacc aacgtcaatc agaggggcct
     5221 gtgtagctac cgataagcgg accctcaaga gggcattagc aatagtgttt ataaggcccc
     5281 cttgttaacc ctaaacgggt agcatatgct tcccgggtag tagtatatac tatccagact
     5341 aaccctaatt caatagcata tgttacccaa cgggaagcat atgctatcga attagggtta
     5401 gtaaaagggt cctaaggaac agcgatatct cccaccccat gagctgtcac ggttttattt
     5461 acatggggtc aggattccac gagggtagtg aaccatttta gtcacaaggg cagtggctga
     5521 agatcaagga gcgggcagtg aactctcctg aatcttcgcc tgcttcttca ttctccttcg
     5581 tttagctaat agaataactg ctgagttgtg aacagtaagg tgtatgtgag gtgctcgaaa
     5641 acaaggtttc aggtgacgcc cccagaataa aatttggacg gggggttcag tggtggcatt
     5701 gtgctatgac accaatataa ccctcacaaa ccccttgggc aataaatact agtgtaggaa
     5761 tgaaacattc tgaatatctt taacaataga aatccatggg gtggggacaa gccgtaaaga
     5821 ctggatgtcc atctcacacg aatttatggc tatgggcaac acataatcct agtgcaatat
     5881 gatactgggg ttattaagat gtgtcccagg cagggaccaa gacaggtgaa ccatgttgtt
     5941 acactctatt tgtaacaagg ggaaagagag tggacgccga cagcagcgga ctccactggt
     6001 tgtctctaac acccccgaaa attaaacggg gctccacgcc aatggggccc ataaacaaag
     6061 acaagtggcc actctttttt ttgaaattgt ggagtggggg cacgcgtcag cccccacacg
     6121 ccgccctgcg gttttggact gtaaaataag ggtgtaataa cttggctgat tgtaaccccg
     6181 ctaaccactg cggtcaaacc acttgcccac aaaaccacta atggcacccc ggggaatacc
     6241 tgcataagta ggtgggcggg ccaagatagg ggcgcgattg ctgcgatctg gaggacaaat
     6301 tacacacact tgcgcctgag cgccaagcac agggttgttg gtcctcatat tcacgaggtc
     6361 gctgagagca cggtgggcta atgttgccat gggtagcata tactacccaa atatctggat
     6421 agcatatgct atcctaatct atatctgggt agcataggct atcctaatct atatctgggt
     6481 agcatatgct atcctaatct atatctgggt agtatatgct atcctaattt atatctgggt
     6541 agcataggct atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt
     6601 agtatatgct atcctaatct gtatccgggt agcatatgct atcctaatag agattagggt
     6661 agtatatgct atcctaattt atatctgggt agcatatact acccaaatat ctggatagca
     6721 tatgctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagca
     6781 taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagta
     6841 tatgctatcc taatttatat ctgggtagca taggctatcc taatctatat ctgggtagca
     6901 tatgctatcc taatctatat ctgggtagta tatgctatcc taatctgtat ccgggtagca
     6961 tatgctatcc tcatgataag ctgtcaaaca tgagaatttt cttgaagacg aaagggcctc
     7021 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     7081 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     7141 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     7201 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     7261 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     7321 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     7381 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     7441 ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     7501 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     7561 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     7621 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     7681 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     7741 acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     7801 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     7861 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     7921 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     7981 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     8041 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     8101 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     8161 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     8221 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     8281 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     8341 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     8401 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     8461 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     8521 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     8581 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     8641 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     8701 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     8761 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     8821 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     8881 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     8941 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     9001 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     9061 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     9121 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     9181 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     9241 aagctctagc tagaggtcga ccaattctca tgtttgacag cttatcatcg cagatccggg
     9301 caacgttgtt gccattgctg caggcgcaga actggtaggt atggaagatc tatacattga
     9361 atcaatattg gcaattagcc atattagtca ttggttatat agcataaatc aatattggct
     9421 attggccatt gcatacgttg tatctatatc ataatatgta catttatatt ggctcatgtc
     9481 caatatgacc gc
//
