LOCUS       sequence-274728-        9354 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 9354)
  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 9354)
  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: D96_Ld_250480
FEATURES             Location/Qualifiers
     source          1..9354
                     /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             4132..4149
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4201..4256
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4255..4274)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6492..6510)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6578..6682
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6683..6751,6752..7543)
                     /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(6901..6920)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7714..8302
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8203..8222
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8456..8473
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8590..8611
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8626..8631,8632..8649,8650..8656)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8664..8680
                     /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     8669..8691
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8688..8704
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8688..8704
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        8975..9354
                     /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 gccgctagag ctgctgctgc aggatctgct
      841 tgtggatctc tggcccacgt gtactacctg cctgacggcc aggatttcag aggcaccgtg
      901 catgtgacag aaggcggcat cccttgccag ctgagaagca gccagacacc taatagccac
      961 gaggtgccac tggattctgc cacaggcgtg ggcaaccaca actactgtag agatgctggc
     1021 ggactgggca gacctggctg ttttgctgct acacctgtgg gcgcctatca gctgtgtgcc
     1081 gttagagccg tgtgtaccgc tagaagcctg gctgccccta cagtgcagtt ttggcctgtg
     1141 tctggaacaa agagagccgc cgacggaagc agctcttacc tggccattag ctgtcacccc
     1201 agaccttgca gcatccacta tgtgctggga gccgaagcca ttgcctctgc cagatctcca
     1261 gtgtacacac agcctctgct gctggaacag agcaccaccg tgaaaaccta cgtcagatac
     1321 gagacagccg agcctatgag agcccaggcc agatatacag tgacccctgc cgctacatct
     1381 accccagccg cctctatcca gtttatccct tccgacgccg tggtgtactc tcaccccatc
     1441 tttgtggaac tgcagggcat caccaatgcc tccgaggtgc tgctgttctg ggccgacgat
     1501 ttcatgaacc ccaccacata caccggcgga gccttcagag tggaaagatc tacatctgtg
     1561 gtggccgtgg tcaacagcac ctacctggtg tctgccagct acaagctgga catcacagcc
     1621 cctccactga acgtgtaccc tccatctggc acatatgtcg gcggcgtgac agtgctcgtt
     1681 agcgatcctc agcctcctgc cacctactat atgtacgtga accagagcac cgactggcag
     1741 tctctgggca cactgctgtt tccatggaca gccatcggca ccagcgagct ggatattaga
     1801 gcactgcacg tggaaggcat cgagagcgcc gagaaggtgg tgtatgaagt ggtgcctgcc
     1861 acacctccta ccatcctgcc tgatgcctct atcgcccacc atagacctgc caacgtgacc
     1921 tgtacagccc cactgggaac accagtggct ctgtctgtgt acagagatgc cgtgatgcag
     1981 gatgccgtgg ccgaaggcgt tttctctgct gtgctttctg agcctggcgc ctacaccgtg
     2041 tcttgcacat atgccgatga cctgcacgcc acacacacca caatggctat tctgcagctg
     2101 gtggctgtgc ccatgcagcc tccaactttt acccctgcct gcggcacctc ttttcccgcc
     2161 attgctctgt ctctgcagat gcagctgaat gtgtccaacc tgctgcctgg cgatctggcc
     2221 acatcttggt ctatttctgc ttctgccggc ggaggcgcca gaattcctat gatcaccaga
     2281 tgcgccgacg acgacggcgt gttcgaatac gatgtgctgc agagcaagag catccaagag
     2341 cctaccacca tggaagtgta cagcaccgct cggagcctga atcctctgga agccgattct
     2401 ccccggacca gctgtaccta ctctctgtat cctctgggcg ccagcgccac accttacttc
     2461 tctgccattc ctgactgtgc caccacatct gccctggcct ctccatcttg tatcatggcc
     2521 ctgaagctcc agctggcctc ctgtctgcac ttctactcta ccgagctgct gagcattgac
     2581 gctctgggac ctatcgtggc catgagaatg acaggactgg ccgctgccat ccagaccgac
     2641 atgtatacca gaatcggcga gtgtctgaca ggcctgcagc atagaggcgt tctgtccgac
     2701 gtgcggaatg agacatctgg cgagatcatg ctggccacaa gctggcatgt ggccactcct
     2761 acaagaatgg ccgctcaggt gtacacagga ctgcccatca cagtccaggt gtccggcttt
     2821 catacagacg ctggcgctta tcacctcgtg cggagcgatt actcttgcga ggatatcgga
     2881 gtgctgcccg aggcagttct ggaaacccct ccatatcatc ctacagccgc cggaagctct
     2941 agagtgtctc ctagcacaga cggcagcaca tacctgacct ttagagtgcc tatcgccggc
     3001 cagtataagc tgtgcagcta catcagcaac gccctgtatg aggtgccctt cagctttagc
     3061 acaggcggag cagctgcacc cgccgtgaat cagtatctgg atgtggttgt gcagccctgg
     3121 ccaacagttg atgccgtgcc tgctgaagaa tgcggaggac tggtgcctcc tgacaaagag
     3181 gatgtcgcct ttagcctgtc tatggccgct gtgccagcag gcgcttatgc cagcctggcc
     3241 tacagcttta atagcgccgg atggggcagc gtgccattcg ctattgatgc tgccacacag
     3301 cttggcggcg ctgcctctct ttctatcggc cctctgaacc ggtacaccag accactggaa
     3361 gtgctggatg cttctctggg ccctacaaca ggcgccgctc agacctgcat ctttttcgtg
     3421 tctgccgcca acagcagcgt gcagagagat gagatcacct acctcttcta caagatgaac
     3481 tccagcgagg tgcccgccga tcttacaggc gttatgctgc tgctccaagg ccagtttcag
     3541 cccgccgaga tgatcgtgat cgacgtgtac gagaacgtga ccctgaatcc aaccaccgcc
     3601 aagaagtcta gcagcctgca gctcctgaga agagtgacag ccagacctag cgccgctaca
     3661 gcatttgctg ctgcccttcc agatgccatc cttggattgc ctgccggcgc tcagcacatc
     3721 ccttatgtga tccatgtgac cctggacggc gtgcacctta cagcagctcc atctactgtg
     3781 gccctgtctc ctctggctct ggccatggaa gcctctacag cttgtcctag cggactggct
     3841 ctgaagggcc agtgcctgtg tagaaacaag cggaccaaag tgatcagcgc ctgcggaacc
     3901 gccatcgagg attctggatc ttctgatgcc ggcgagcctc agtctagaag ctctgccgat
     3961 agctctagct gctccaccac cgatagcagc gacgacaccg ataccaagac agtgtccctg
     4021 ggacagagac tggctggcgc gcccggcagc ctgcaccaca tcctggacgc ccagaagatg
     4081 ctgtggaacc acagggacag gaacctgcct cctctggccc ctctgggccc ccaccaccat
     4141 caccaccatt gagtctagac acgtgattaa ttaaggatcc cccgacctcg acctctggct
     4201 aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg
     4261 gaaggacata tgggagggca aatcatttgg tcgagatccc tcggagatct ctagctagag
     4321 gatcgatccc cgccccggac gaactaaacc tgactacgac atctctgccc cttcttcgcg
     4381 gggcagtgca tgtaatccct tcagttggtt ggtacaactt gccaactggg ccctgttcca
     4441 catgtgacac ggggggggac caaacacaaa ggggttctct gactgtagtt gacatcctta
     4501 taaatggatg tgcacatttg ccaacactga gtggctttca tcctggagca gactttgcag
     4561 tctgtggact gcaacacaac attgccttta tgtgtaactc ttggctgaag ctcttacacc
     4621 aatgctgggg gacatgtacc tcccaggggc ccaggaagac tacgggaggc tacaccaacg
     4681 tcaatcagag gggcctgtgt agctaccgat aagcggaccc tcaagagggc attagcaata
     4741 gtgtttataa ggcccccttg ttaaccctaa acgggtagca tatgcttccc gggtagtagt
     4801 atatactatc cagactaacc ctaattcaat agcatatgtt acccaacggg aagcatatgc
     4861 tatcgaatta gggttagtaa aagggtccta aggaacagcg atatctccca ccccatgagc
     4921 tgtcacggtt ttatttacat ggggtcagga ttccacgagg gtagtgaacc attttagtca
     4981 caagggcagt ggctgaagat caaggagcgg gcagtgaact ctcctgaatc ttcgcctgct
     5041 tcttcattct ccttcgttta gctaatagaa taactgctga gttgtgaaca gtaaggtgta
     5101 tgtgaggtgc tcgaaaacaa ggtttcaggt gacgccccca gaataaaatt tggacggggg
     5161 gttcagtggt ggcattgtgc tatgacacca atataaccct cacaaacccc ttgggcaata
     5221 aatactagtg taggaatgaa acattctgaa tatctttaac aatagaaatc catggggtgg
     5281 ggacaagccg taaagactgg atgtccatct cacacgaatt tatggctatg ggcaacacat
     5341 aatcctagtg caatatgata ctggggttat taagatgtgt cccaggcagg gaccaagaca
     5401 ggtgaaccat gttgttacac tctatttgta acaaggggaa agagagtgga cgccgacagc
     5461 agcggactcc actggttgtc tctaacaccc ccgaaaatta aacggggctc cacgccaatg
     5521 gggcccataa acaaagacaa gtggccactc ttttttttga aattgtggag tgggggcacg
     5581 cgtcagcccc cacacgccgc cctgcggttt tggactgtaa aataagggtg taataacttg
     5641 gctgattgta accccgctaa ccactgcggt caaaccactt gcccacaaaa ccactaatgg
     5701 caccccgggg aatacctgca taagtaggtg ggcgggccaa gataggggcg cgattgctgc
     5761 gatctggagg acaaattaca cacacttgcg cctgagcgcc aagcacaggg ttgttggtcc
     5821 tcatattcac gaggtcgctg agagcacggt gggctaatgt tgccatgggt agcatatact
     5881 acccaaatat ctggatagca tatgctatcc taatctatat ctgggtagca taggctatcc
     5941 taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagta tatgctatcc
     6001 taatttatat ctgggtagca taggctatcc taatctatat ctgggtagca tatgctatcc
     6061 taatctatat ctgggtagta tatgctatcc taatctgtat ccgggtagca tatgctatcc
     6121 taatagagat tagggtagta tatgctatcc taatttatat ctgggtagca tatactaccc
     6181 aaatatctgg atagcatatg ctatcctaat ctatatctgg gtagcatatg ctatcctaat
     6241 ctatatctgg gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat
     6301 ctatatctgg gtagtatatg ctatcctaat ttatatctgg gtagcatagg ctatcctaat
     6361 ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat
     6421 ctgtatccgg gtagcatatg ctatcctcat gataagctgt caaacatgag aattttcttg
     6481 aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     6541 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     6601 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     6661 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     6721 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     6781 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     6841 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     6901 gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat
     6961 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     7021 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     7081 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     7141 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     7201 cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac
     7261 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     7321 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     7381 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     7441 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     7501 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     7561 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     7621 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     7681 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     7741 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     7801 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     7861 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     7921 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     7981 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     8041 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     8101 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     8161 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     8221 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     8281 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     8341 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     8401 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     8461 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     8521 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     8581 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     8641 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
     8701 tgaccatgat tacgccaagc tctagctaga ggtcgaccaa ttctcatgtt tgacagctta
     8761 tcatcgcaga tccgggcaac gttgttgcca ttgctgcagg cgcagaactg gtaggtatgg
     8821 aagatctata cattgaatca atattggcaa ttagccatat tagtcattgg ttatatagca
     8881 taaatcaata ttggctattg gccattgcat acgttgtatc tatatcataa tatgtacatt
     8941 tatattggct catgtccaat atgaccgcca tgttgacatt gattattgac tagttattaa
     9001 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     9061 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     9121 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     9181 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtccgccc
     9241 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     9301 cgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catg
//
