LOCUS       sequence-234473-        7011 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 7011)
  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 7011)
  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_105450-Cd4-bio-His
FEATURES             Location/Qualifiers
     source          1..7011
                     /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             1789..1806
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1858..1913
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1912..1931)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4149..4167)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4235..4339
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4340..4408,4409..5200)
                     /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(4558..4577)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5371..5959
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5860..5879
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6113..6130
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6247..6268
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6283..6288,6289..6306,6307..6313)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6321..6337
                     /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     6326..6348
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6345..6361
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6345..6361
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        6632..7011
                     /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 gcctacagcg tgaagaacgt ggacatgagc
      841 aacaccaaag aaaccgtgga ctaccagctg gacaacaaca gcacctgtag catgtgcatc
      901 cagttcatca ccaagtggca gacctacctg aacagcacca gcgtggacga gaagatcgag
      961 cagttcatcc ggtctacctg cagcttctac ttcttcttcc ggtatcgctg cgaggaattc
     1021 atggaacgct acgtgaacaa gaccgtgtac atcatccagc acaacaacgc caccaccctg
     1081 tgcaagttca gcatcatcga gtgcatgctg atcatcctga agctgacccc cctgtgtggc
     1141 gcgccgtcga cctccatcac ggcctataag agtgaggggg agtcagcgga gttctccttc
     1201 ccactcaacc ttggagagga aagcctgcag ggagagttga gatggaaggc agagaaggct
     1261 ccttcttccc agtcctggat caccttctcc ctaaagaacc aaaaggtgtc tgtgcagaag
     1321 tctactagca accccaagtt ccagctgtcc gaaacgctcc cactcaccct tcagataccc
     1381 caggtctccc ttcagtttgc tggttctggc aacctgaccc tgactctgga cagagggata
     1441 ctgtatcagg aagtgaacct ggtggtgatg aaagtgactc agcccgacag caacactttg
     1501 acctgtgagg tgatgggacc cacctcaccc aagatgagac tgatcttgaa gcaggagaat
     1561 caggaggcca gggtctccag gcaggagaaa gtgattcaag tgcaggcccc tgaagcaggg
     1621 gtgtggcaat gtctactgag tgaaggtgaa gaggtcaaga tggactccaa gatccaggtt
     1681 ttatccaaag ggttgaattc cggcagcctg caccacatcc tggacgccca gaagatgctg
     1741 tggaaccaca gggacaggaa cctgcctcct ctggcccctc tgggccccca ccaccatcac
     1801 caccattgag tctagacacg tgattaatta aggatccccc gacctcgacc tctggctaat
     1861 aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa
     1921 ggacatatgg gagggcaaat catttggtcg agatccctcg gagatctcta gctagaggat
     1981 cgatccccgc cccggacgaa ctaaacctga ctacgacatc tctgcccctt cttcgcgggg
     2041 cagtgcatgt aatcccttca gttggttggt acaacttgcc aactgggccc tgttccacat
     2101 gtgacacggg gggggaccaa acacaaaggg gttctctgac tgtagttgac atccttataa
     2161 atggatgtgc acatttgcca acactgagtg gctttcatcc tggagcagac tttgcagtct
     2221 gtggactgca acacaacatt gcctttatgt gtaactcttg gctgaagctc ttacaccaat
     2281 gctgggggac atgtacctcc caggggccca ggaagactac gggaggctac accaacgtca
     2341 atcagagggg cctgtgtagc taccgataag cggaccctca agagggcatt agcaatagtg
     2401 tttataaggc ccccttgtta accctaaacg ggtagcatat gcttcccggg tagtagtata
     2461 tactatccag actaacccta attcaatagc atatgttacc caacgggaag catatgctat
     2521 cgaattaggg ttagtaaaag ggtcctaagg aacagcgata tctcccaccc catgagctgt
     2581 cacggtttta tttacatggg gtcaggattc cacgagggta gtgaaccatt ttagtcacaa
     2641 gggcagtggc tgaagatcaa ggagcgggca gtgaactctc ctgaatcttc gcctgcttct
     2701 tcattctcct tcgtttagct aatagaataa ctgctgagtt gtgaacagta aggtgtatgt
     2761 gaggtgctcg aaaacaaggt ttcaggtgac gcccccagaa taaaatttgg acggggggtt
     2821 cagtggtggc attgtgctat gacaccaata taaccctcac aaaccccttg ggcaataaat
     2881 actagtgtag gaatgaaaca ttctgaatat ctttaacaat agaaatccat ggggtgggga
     2941 caagccgtaa agactggatg tccatctcac acgaatttat ggctatgggc aacacataat
     3001 cctagtgcaa tatgatactg gggttattaa gatgtgtccc aggcagggac caagacaggt
     3061 gaaccatgtt gttacactct atttgtaaca aggggaaaga gagtggacgc cgacagcagc
     3121 ggactccact ggttgtctct aacacccccg aaaattaaac ggggctccac gccaatgggg
     3181 cccataaaca aagacaagtg gccactcttt tttttgaaat tgtggagtgg gggcacgcgt
     3241 cagcccccac acgccgccct gcggttttgg actgtaaaat aagggtgtaa taacttggct
     3301 gattgtaacc ccgctaacca ctgcggtcaa accacttgcc cacaaaacca ctaatggcac
     3361 cccggggaat acctgcataa gtaggtgggc gggccaagat aggggcgcga ttgctgcgat
     3421 ctggaggaca aattacacac acttgcgcct gagcgccaag cacagggttg ttggtcctca
     3481 tattcacgag gtcgctgaga gcacggtggg ctaatgttgc catgggtagc atatactacc
     3541 caaatatctg gatagcatat gctatcctaa tctatatctg ggtagcatag gctatcctaa
     3601 tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagtatat gctatcctaa
     3661 tttatatctg ggtagcatag gctatcctaa tctatatctg ggtagcatat gctatcctaa
     3721 tctatatctg ggtagtatat gctatcctaa tctgtatccg ggtagcatat gctatcctaa
     3781 tagagattag ggtagtatat gctatcctaa tttatatctg ggtagcatat actacccaaa
     3841 tatctggata gcatatgcta tcctaatcta tatctgggta gcatatgcta tcctaatcta
     3901 tatctgggta gcataggcta tcctaatcta tatctgggta gcatatgcta tcctaatcta
     3961 tatctgggta gtatatgcta tcctaattta tatctgggta gcataggcta tcctaatcta
     4021 tatctgggta gcatatgcta tcctaatcta tatctgggta gtatatgcta tcctaatctg
     4081 tatccgggta gcatatgcta tcctcatgat aagctgtcaa acatgagaat tttcttgaag
     4141 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     4201 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     4261 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     4321 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     4381 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     4441 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     4501 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     4561 atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca
     4621 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     4681 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     4741 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     4801 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     4861 cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg
     4921 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     4981 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     5041 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     5101 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     5161 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     5221 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     5281 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     5341 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5401 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5461 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     5521 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5581 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5641 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     5701 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     5761 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     5821 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     5881 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     5941 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6001 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     6061 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     6121 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     6181 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     6241 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     6301 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     6361 ccatgattac gccaagctct agctagaggt cgaccaattc tcatgtttga cagcttatca
     6421 tcgcagatcc gggcaacgtt gttgccattg ctgcaggcgc agaactggta ggtatggaag
     6481 atctatacat tgaatcaata ttggcaatta gccatattag tcattggtta tatagcataa
     6541 atcaatattg gctattggcc attgcatacg ttgtatctat atcataatat gtacatttat
     6601 attggctcat gtccaatatg accgccatgt tgacattgat tattgactag ttattaatag
     6661 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
     6721 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     6781 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     6841 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct
     6901 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg
     6961 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat g
//
