LOCUS       sequence-234779-        7410 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of recombinant S. mansoni protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7410)
  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 7410)
  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: "bioRXiv"
COMMENT     Sequence Label: Smp_061310-Cd4-bio-His
FEATURES             Location/Qualifiers
     source          1..7410
                     /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             2188..2205
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2257..2312
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2311..2330)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4548..4566)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4634..4738
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4739..4807,4808..5599)
                     /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(4957..4976)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5770..6358
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6259..6278
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6512..6529
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6646..6667
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6682..6687,6688..6705,6706..6712)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6720..6736
                     /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     6725..6747
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6744..6760
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6744..6760
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7031..7410
                     /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 gcctatgtga tcgagggcac cgtggtcatg
      841 ccccccgagg cccctccaga ctggcacgtg aacacccgga tcaatatcgc cggcgaccag
      901 ttcgtgggct tcgtgaaggc cgacggcaac ttcgaagtga ccaacgtgcc cagcggcagc
      961 tacatcgtgg aagtgatcaa ccccgtgtac tattttcagg gcgtgcgggt ggacatcagc
     1021 agcaagggcc ggatccgggc cagacagctg aacgcccctc agcccaacgc cgtgaaagag
     1081 ctgccctacc ccctgcggct gagcagcagc ggcagagccg tgtacttcaa ggccagagag
     1141 cagctgcgga ccctggacct gctgttcaac cccaacgtgc tgtacgtgct ggtgcccctg
     1201 ctgctggtgg tcatcctgac caagctggtc aacaccaacg accccagcgt gcagaaagaa
     1261 atgcagcaga tgaacatcct gaacccccag cagcagctgc ccgatatgag cgagttcctg
     1321 agcagcctga gcctgttcgg cggcaacacc aaaaagacca accccaccac catcaacaag
     1381 agaaagagcg ccgagaagca ccggcaggtc acaaacagcg ccggcatcag cgagagcccc
     1441 agccagtaca ccaccgtgtc caacaccagc aacaccacca gctccagcag caacagcgcc
     1501 agcaagcgga ccaccaccaa ggcccggaag gccgaaggcg cgccgtcgac ctccatcacg
     1561 gcctataaga gtgaggggga gtcagcggag ttctccttcc cactcaacct tggagaggaa
     1621 agcctgcagg gagagttgag atggaaggca gagaaggctc cttcttccca gtcctggatc
     1681 accttctccc taaagaacca aaaggtgtct gtgcagaagt ctactagcaa ccccaagttc
     1741 cagctgtccg aaacgctccc actcaccctt cagatacccc aggtctccct tcagtttgct
     1801 ggttctggca acctgaccct gactctggac agagggatac tgtatcagga agtgaacctg
     1861 gtggtgatga aagtgactca gcccgacagc aacactttga cctgtgaggt gatgggaccc
     1921 acctcaccca agatgagact gatcttgaag caggagaatc aggaggccag ggtctccagg
     1981 caggagaaag tgattcaagt gcaggcccct gaagcagggg tgtggcaatg tctactgagt
     2041 gaaggtgaag aggtcaagat ggactccaag atccaggttt tatccaaagg gttgaattcc
     2101 ggcagcctgc accacatcct ggacgcccag aagatgctgt ggaaccacag ggacaggaac
     2161 ctgcctcctc tggcccctct gggcccccac caccatcacc accattgagt ctagacacgt
     2221 gattaattaa ggatcccccg acctcgacct ctggctaata aaggaaattt attttcattg
     2281 caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc
     2341 atttggtcga gatccctcgg agatctctag ctagaggatc gatccccgcc ccggacgaac
     2401 taaacctgac tacgacatct ctgccccttc ttcgcggggc agtgcatgta atcccttcag
     2461 ttggttggta caacttgcca actgggccct gttccacatg tgacacgggg ggggaccaaa
     2521 cacaaagggg ttctctgact gtagttgaca tccttataaa tggatgtgca catttgccaa
     2581 cactgagtgg ctttcatcct ggagcagact ttgcagtctg tggactgcaa cacaacattg
     2641 cctttatgtg taactcttgg ctgaagctct tacaccaatg ctgggggaca tgtacctccc
     2701 aggggcccag gaagactacg ggaggctaca ccaacgtcaa tcagaggggc ctgtgtagct
     2761 accgataagc ggaccctcaa gagggcatta gcaatagtgt ttataaggcc cccttgttaa
     2821 ccctaaacgg gtagcatatg cttcccgggt agtagtatat actatccaga ctaaccctaa
     2881 ttcaatagca tatgttaccc aacgggaagc atatgctatc gaattagggt tagtaaaagg
     2941 gtcctaagga acagcgatat ctcccacccc atgagctgtc acggttttat ttacatgggg
     3001 tcaggattcc acgagggtag tgaaccattt tagtcacaag ggcagtggct gaagatcaag
     3061 gagcgggcag tgaactctcc tgaatcttcg cctgcttctt cattctcctt cgtttagcta
     3121 atagaataac tgctgagttg tgaacagtaa ggtgtatgtg aggtgctcga aaacaaggtt
     3181 tcaggtgacg cccccagaat aaaatttgga cggggggttc agtggtggca ttgtgctatg
     3241 acaccaatat aaccctcaca aaccccttgg gcaataaata ctagtgtagg aatgaaacat
     3301 tctgaatatc tttaacaata gaaatccatg gggtggggac aagccgtaaa gactggatgt
     3361 ccatctcaca cgaatttatg gctatgggca acacataatc ctagtgcaat atgatactgg
     3421 ggttattaag atgtgtccca ggcagggacc aagacaggtg aaccatgttg ttacactcta
     3481 tttgtaacaa ggggaaagag agtggacgcc gacagcagcg gactccactg gttgtctcta
     3541 acacccccga aaattaaacg gggctccacg ccaatggggc ccataaacaa agacaagtgg
     3601 ccactctttt ttttgaaatt gtggagtggg ggcacgcgtc agcccccaca cgccgccctg
     3661 cggttttgga ctgtaaaata agggtgtaat aacttggctg attgtaaccc cgctaaccac
     3721 tgcggtcaaa ccacttgccc acaaaaccac taatggcacc ccggggaata cctgcataag
     3781 taggtgggcg ggccaagata ggggcgcgat tgctgcgatc tggaggacaa attacacaca
     3841 cttgcgcctg agcgccaagc acagggttgt tggtcctcat attcacgagg tcgctgagag
     3901 cacggtgggc taatgttgcc atgggtagca tatactaccc aaatatctgg atagcatatg
     3961 ctatcctaat ctatatctgg gtagcatagg ctatcctaat ctatatctgg gtagcatatg
     4021 ctatcctaat ctatatctgg gtagtatatg ctatcctaat ttatatctgg gtagcatagg
     4081 ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagtatatg
     4141 ctatcctaat ctgtatccgg gtagcatatg ctatcctaat agagattagg gtagtatatg
     4201 ctatcctaat ttatatctgg gtagcatata ctacccaaat atctggatag catatgctat
     4261 cctaatctat atctgggtag catatgctat cctaatctat atctgggtag cataggctat
     4321 cctaatctat atctgggtag catatgctat cctaatctat atctgggtag tatatgctat
     4381 cctaatttat atctgggtag cataggctat cctaatctat atctgggtag catatgctat
     4441 cctaatctat atctgggtag tatatgctat cctaatctgt atccgggtag catatgctat
     4501 cctcatgata agctgtcaaa catgagaatt ttcttgaaga cgaaagggcc tcgtgatacg
     4561 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     4621 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     4681 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     4741 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     4801 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     4861 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     4921 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     4981 tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     5041 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     5101 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     5161 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     5221 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     5281 tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     5341 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     5401 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     5461 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     5521 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     5581 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     5641 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     5701 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     5761 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     5821 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     5881 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     5941 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     6001 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     6061 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     6121 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     6181 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     6241 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     6301 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     6361 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     6421 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     6481 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     6541 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     6601 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
     6661 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
     6721 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagctcta
     6781 gctagaggtc gaccaattct catgtttgac agcttatcat cgcagatccg ggcaacgttg
     6841 ttgccattgc tgcaggcgca gaactggtag gtatggaaga tctatacatt gaatcaatat
     6901 tggcaattag ccatattagt cattggttat atagcataaa tcaatattgg ctattggcca
     6961 ttgcatacgt tgtatctata tcataatatg tacatttata ttggctcatg tccaatatga
     7021 ccgccatgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
     7081 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
     7141 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     7201 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     7261 gcagtacatc aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa
     7321 tggcccgcct ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac
     7381 atctacgtat tagtcatcgc tattaccatg
//
