LOCUS       sequence-274664-        6618 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 6618)
  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 6618)
  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: D56_Ld_141480
FEATURES             Location/Qualifiers
     source          1..6618
                     /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             1776..1793
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1845..1900
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1899..1918)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4136..4154)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4222..4326
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4327..4395,4396..5187)
                     /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(4545..4564)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5358..5946
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5847..5866
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6100..6117
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6234..6255
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6270..6275,6276..6293,6294..6300)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6308..6324
                     /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     6313..6335
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6332..6348
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6332..6348
                     /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 gccgtgctgc
     1201 ctgctacagc cctgagagtg aagcccacca tcaccgccct gaccttcgac gtgggcaaag
     1261 aggaagtctg cctgtacagc atgggcaagg acttctacga gggcgtgacc atgcactacc
     1321 acgtgatgga aggcgagaac gacttcgacg tgttcatccg ggacgtggac ggccacgtga
     1381 tctacgccag ctatgccggc gagcacggcg ccgaggaccg ggtgtacttt accacccacg
     1441 ccaacaaaga gcacgcctac tgcatcgaca accgggacta cagcggcgag ctgaaagtga
     1501 tcaagatgga agtgggcctg accagcctga agcggtggaa gaaccggatc gaccccctgc
     1561 ggaagctgat gacccggacc gacggcttcg tgatgggcat gcacgacgac cagatcctgc
     1621 tgcggatgaa ggaacagggc atccgcgagt gggtggaaaa gatcttcacc atgctgacag
     1681 gcgcgcccgg cagcctgcac cacatcctgg acgcccagaa gatgctgtgg aaccacaggg
     1741 acaggaacct gcctcctctg gcccctctgg gcccccacca ccatcaccac cattgagtct
     1801 agacacgtga ttaattaagg atcccccgac ctcgacctct ggctaataaa ggaaatttat
     1861 tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag
     1921 ggcaaatcat ttggtcgaga tccctcggag atctctagct agaggatcga tccccgcccc
     1981 ggacgaacta aacctgacta cgacatctct gccccttctt cgcggggcag tgcatgtaat
     2041 cccttcagtt ggttggtaca acttgccaac tgggccctgt tccacatgtg acacgggggg
     2101 ggaccaaaca caaaggggtt ctctgactgt agttgacatc cttataaatg gatgtgcaca
     2161 tttgccaaca ctgagtggct ttcatcctgg agcagacttt gcagtctgtg gactgcaaca
     2221 caacattgcc tttatgtgta actcttggct gaagctctta caccaatgct gggggacatg
     2281 tacctcccag gggcccagga agactacggg aggctacacc aacgtcaatc agaggggcct
     2341 gtgtagctac cgataagcgg accctcaaga gggcattagc aatagtgttt ataaggcccc
     2401 cttgttaacc ctaaacgggt agcatatgct tcccgggtag tagtatatac tatccagact
     2461 aaccctaatt caatagcata tgttacccaa cgggaagcat atgctatcga attagggtta
     2521 gtaaaagggt cctaaggaac agcgatatct cccaccccat gagctgtcac ggttttattt
     2581 acatggggtc aggattccac gagggtagtg aaccatttta gtcacaaggg cagtggctga
     2641 agatcaagga gcgggcagtg aactctcctg aatcttcgcc tgcttcttca ttctccttcg
     2701 tttagctaat agaataactg ctgagttgtg aacagtaagg tgtatgtgag gtgctcgaaa
     2761 acaaggtttc aggtgacgcc cccagaataa aatttggacg gggggttcag tggtggcatt
     2821 gtgctatgac accaatataa ccctcacaaa ccccttgggc aataaatact agtgtaggaa
     2881 tgaaacattc tgaatatctt taacaataga aatccatggg gtggggacaa gccgtaaaga
     2941 ctggatgtcc atctcacacg aatttatggc tatgggcaac acataatcct agtgcaatat
     3001 gatactgggg ttattaagat gtgtcccagg cagggaccaa gacaggtgaa ccatgttgtt
     3061 acactctatt tgtaacaagg ggaaagagag tggacgccga cagcagcgga ctccactggt
     3121 tgtctctaac acccccgaaa attaaacggg gctccacgcc aatggggccc ataaacaaag
     3181 acaagtggcc actctttttt ttgaaattgt ggagtggggg cacgcgtcag cccccacacg
     3241 ccgccctgcg gttttggact gtaaaataag ggtgtaataa cttggctgat tgtaaccccg
     3301 ctaaccactg cggtcaaacc acttgcccac aaaaccacta atggcacccc ggggaatacc
     3361 tgcataagta ggtgggcggg ccaagatagg ggcgcgattg ctgcgatctg gaggacaaat
     3421 tacacacact tgcgcctgag cgccaagcac agggttgttg gtcctcatat tcacgaggtc
     3481 gctgagagca cggtgggcta atgttgccat gggtagcata tactacccaa atatctggat
     3541 agcatatgct atcctaatct atatctgggt agcataggct atcctaatct atatctgggt
     3601 agcatatgct atcctaatct atatctgggt agtatatgct atcctaattt atatctgggt
     3661 agcataggct atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt
     3721 agtatatgct atcctaatct gtatccgggt agcatatgct atcctaatag agattagggt
     3781 agtatatgct atcctaattt atatctgggt agcatatact acccaaatat ctggatagca
     3841 tatgctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagca
     3901 taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagta
     3961 tatgctatcc taatttatat ctgggtagca taggctatcc taatctatat ctgggtagca
     4021 tatgctatcc taatctatat ctgggtagta tatgctatcc taatctgtat ccgggtagca
     4081 tatgctatcc tcatgataag ctgtcaaaca tgagaatttt cttgaagacg aaagggcctc
     4141 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     4201 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     4261 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     4321 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     4381 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     4441 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     4501 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     4561 ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     4621 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     4681 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     4741 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     4801 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     4861 acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     4921 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     4981 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     5041 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     5101 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     5161 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     5221 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     5281 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     5341 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     5401 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     5461 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     5521 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     5581 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     5641 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     5701 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     5761 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     5821 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     5881 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     5941 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     6001 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     6061 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     6121 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     6181 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     6241 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     6301 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     6361 aagctctagc tagaggtcga ccaattctca tgtttgacag cttatcatcg cagatccggg
     6421 caacgttgtt gccattgctg caggcgcaga actggtaggt atggaagatc tatacattga
     6481 atcaatattg gcaattagcc atattagtca ttggttatat agcataaatc aatattggct
     6541 attggccatt gcatacgttg tatctatatc ataatatgta catttatatt ggctcatgtc
     6601 caatatgacc gccatgtt
//
