LOCUS       sequence-234767-        7056 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expression of recombinant S. mansoni protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7056)
  AUTHORS   Cecile Crosnier, Anna V. Protasio, Gabriel Rinaldi, Shona Wilson, 
            Matthew Berriman, Gavin J. Wright
  TITLE     A library of cell-surface and secreted proteins from Schistosoma 
            mansoni identifies early serological markers of infection
  JOURNAL   bioRXiv
REFERENCE   2  (bases 1 to 7056)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRXiv"
COMMENT     Sequence Label: Smp_138060-Cd4-bio-His
FEATURES             Location/Qualifiers
     source          1..7056
                     /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             2214..2231
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2283..2338
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2337..2356)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4574..4592)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4660..4764
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4765..4833,4834..5625)
                     /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(4983..5002)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5796..6384
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6285..6304
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6538..6555
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6672..6693
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6708..6713,6714..6731,6732..6738)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6746..6762
                     /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     6751..6773
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6770..6786
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6770..6786
                     /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 gccgctcagc
     1201 aggaatgcga gaagaactgc aagggcgaca acgagtacgt gtcccccaac tgcggcatcc
     1261 tgtgcagcgg caccatcggc ccccagacct tctactgcta cctgggctgc agccacaacg
     1321 ccaccaagca gagcgagttc gacaactgca agaccaagtg cgacggcggc gtgcagctga
     1381 ccaaagaggc ctgcctgagc aactgcggcc tgatcaccac ccaccccgag ctgtgtgatg
     1441 ccgtgtgcgg cggcagcctg tacaactgcg accagaagca caccgacccc cggaaggatg
     1501 gcgccgacgg cagcgaggac ttcgataagt gcaagacaaa atgctacaag atggccggcc
     1561 agggcgcgcc gtcgacctcc atcacggcct ataagagtga gggggagtca gcggagttct
     1621 ccttcccact caaccttgga gaggaaagcc tgcagggaga gttgagatgg aaggcagaga
     1681 aggctccttc ttcccagtcc tggatcacct tctccctaaa gaaccaaaag gtgtctgtgc
     1741 agaagtctac tagcaacccc aagttccagc tgtccgaaac gctcccactc acccttcaga
     1801 taccccaggt ctcccttcag tttgctggtt ctggcaacct gaccctgact ctggacagag
     1861 ggatactgta tcaggaagtg aacctggtgg tgatgaaagt gactcagccc gacagcaaca
     1921 ctttgacctg tgaggtgatg ggacccacct cacccaagat gagactgatc ttgaagcagg
     1981 agaatcagga ggccagggtc tccaggcagg agaaagtgat tcaagtgcag gcccctgaag
     2041 caggggtgtg gcaatgtcta ctgagtgaag gtgaagaggt caagatggac tccaagatcc
     2101 aggttttatc caaagggttg aattccggca gcctgcacca catcctggac gcccagaaga
     2161 tgctgtggaa ccacagggac aggaacctgc ctcctctggc ccctctgggc ccccaccacc
     2221 atcaccacca ttgagtctag acacgtgatt aattaaggat cccccgacct cgacctctgg
     2281 ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact
     2341 cggaaggaca tatgggaggg caaatcattt ggtcgagatc cctcggagat ctctagctag
     2401 aggatcgatc cccgccccgg acgaactaaa cctgactacg acatctctgc cccttcttcg
     2461 cggggcagtg catgtaatcc cttcagttgg ttggtacaac ttgccaactg ggccctgttc
     2521 cacatgtgac acgggggggg accaaacaca aaggggttct ctgactgtag ttgacatcct
     2581 tataaatgga tgtgcacatt tgccaacact gagtggcttt catcctggag cagactttgc
     2641 agtctgtgga ctgcaacaca acattgcctt tatgtgtaac tcttggctga agctcttaca
     2701 ccaatgctgg gggacatgta cctcccaggg gcccaggaag actacgggag gctacaccaa
     2761 cgtcaatcag aggggcctgt gtagctaccg ataagcggac cctcaagagg gcattagcaa
     2821 tagtgtttat aaggccccct tgttaaccct aaacgggtag catatgcttc ccgggtagta
     2881 gtatatacta tccagactaa ccctaattca atagcatatg ttacccaacg ggaagcatat
     2941 gctatcgaat tagggttagt aaaagggtcc taaggaacag cgatatctcc caccccatga
     3001 gctgtcacgg ttttatttac atggggtcag gattccacga gggtagtgaa ccattttagt
     3061 cacaagggca gtggctgaag atcaaggagc gggcagtgaa ctctcctgaa tcttcgcctg
     3121 cttcttcatt ctccttcgtt tagctaatag aataactgct gagttgtgaa cagtaaggtg
     3181 tatgtgaggt gctcgaaaac aaggtttcag gtgacgcccc cagaataaaa tttggacggg
     3241 gggttcagtg gtggcattgt gctatgacac caatataacc ctcacaaacc ccttgggcaa
     3301 taaatactag tgtaggaatg aaacattctg aatatcttta acaatagaaa tccatggggt
     3361 ggggacaagc cgtaaagact ggatgtccat ctcacacgaa tttatggcta tgggcaacac
     3421 ataatcctag tgcaatatga tactggggtt attaagatgt gtcccaggca gggaccaaga
     3481 caggtgaacc atgttgttac actctatttg taacaagggg aaagagagtg gacgccgaca
     3541 gcagcggact ccactggttg tctctaacac ccccgaaaat taaacggggc tccacgccaa
     3601 tggggcccat aaacaaagac aagtggccac tctttttttt gaaattgtgg agtgggggca
     3661 cgcgtcagcc cccacacgcc gccctgcggt tttggactgt aaaataaggg tgtaataact
     3721 tggctgattg taaccccgct aaccactgcg gtcaaaccac ttgcccacaa aaccactaat
     3781 ggcaccccgg ggaatacctg cataagtagg tgggcgggcc aagatagggg cgcgattgct
     3841 gcgatctgga ggacaaatta cacacacttg cgcctgagcg ccaagcacag ggttgttggt
     3901 cctcatattc acgaggtcgc tgagagcacg gtgggctaat gttgccatgg gtagcatata
     3961 ctacccaaat atctggatag catatgctat cctaatctat atctgggtag cataggctat
     4021 cctaatctat atctgggtag catatgctat cctaatctat atctgggtag tatatgctat
     4081 cctaatttat atctgggtag cataggctat cctaatctat atctgggtag catatgctat
     4141 cctaatctat atctgggtag tatatgctat cctaatctgt atccgggtag catatgctat
     4201 cctaatagag attagggtag tatatgctat cctaatttat atctgggtag catatactac
     4261 ccaaatatct ggatagcata tgctatccta atctatatct gggtagcata tgctatccta
     4321 atctatatct gggtagcata ggctatccta atctatatct gggtagcata tgctatccta
     4381 atctatatct gggtagtata tgctatccta atttatatct gggtagcata ggctatccta
     4441 atctatatct gggtagcata tgctatccta atctatatct gggtagtata tgctatccta
     4501 atctgtatcc gggtagcata tgctatcctc atgataagct gtcaaacatg agaattttct
     4561 tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     4621 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     4681 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     4741 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     4801 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     4861 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     4921 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     4981 ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc
     5041 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     5101 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     5161 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     5221 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     5281 aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta
     5341 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     5401 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     5461 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     5521 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     5581 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     5641 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     5701 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     5761 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     5821 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     5881 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     5941 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6001 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6061 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6121 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6181 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6241 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6301 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6361 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6421 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     6481 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     6541 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     6601 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     6661 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     6721 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     6781 tatgaccatg attacgccaa gctctagcta gaggtcgacc aattctcatg tttgacagct
     6841 tatcatcgca gatccgggca acgttgttgc cattgctgca ggcgcagaac tggtaggtat
     6901 ggaagatcta tacattgaat caatattggc aattagccat attagtcatt ggttatatag
     6961 cataaatcaa tattggctat tggccattgc atacgttgta tctatatcat aatatgtaca
     7021 tttatattgg ctcatgtcca atatgaccgc catgtt
//
