LOCUS       sequence-229244-        7164 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 7164)
  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 7164)
  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_063330-Cd4-bio-His
FEATURES             Location/Qualifiers
     source          1..7164
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        7..386
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        387..590
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     541..561
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     587..611
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     1092..1111
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1134..1143
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2328..2345
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2397..2452
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2451..2470)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4688..4706)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4774..4878
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4879..4947,4948..5739)
                     /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(5097..5116)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5910..6498
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6399..6418
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6652..6669
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6786..6807
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6822..6827,6828..6845,6846..6852)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6860..6876
                     /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     6865..6887
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6884..6900
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6884..6900
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 catgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
       61 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      121 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      181 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      241 tacatcaagt gtatcatatg ccaagtccgc cccctattga cgtcaatgac ggtaaatggc
      301 ccgcctggca ttatgcccag tacatgacct tacgggactt tcctacttgg cagtacatct
      361 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacacc aatgggcgtg
      421 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      481 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taataacccc gccccgttga
      541 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga
      601 accgtcagat cctcactctc ttccgcatcg ctgtctgcga gggccagctg ttgggctcgc
      661 ggttgaggac aaactcttcg cggtctttcc agtactcttg gatcggaaac ccgtcggcct
      721 ccgaacggta ctccgccacc gagggacctg agcgagtccg catcgaccgg atcggaaaac
      781 ctctcgagaa aggcgtctaa ccagtcacag tcgcaaggta ggctgagcac cgtggcgggc
      841 ggcagcgggt ggcggtcggg gttgtttctg gcggaggtgc tgctgatgat gtaattaaag
      901 taggcggtct tgagacggcg gatggtcgag gtgaggtgtg gcaggcttga gatccagctg
      961 ttggggtgag tactccctct caaaagcggg cattacttct gcgctaagat tgtcagtttc
     1021 caaaaacgag gaggatttga tattcacctg gcccgatctg gccatacact tgagtgacaa
     1081 tgacatccac tttgcctttc tctccacagg tgtccactcc caggtccaag tttgccacca
     1141 tggagtttca gacccaggta ctcatgtccc tgctgctctg catgtctggt gcggccgcca
     1201 agcctgtgca cctgaagtcc gagacaagcg agagcggcga cgagctgctg gacatcaaca
     1261 aggacgacca gtacagcgac tacgacgagc tgaagcggat cttccaggaa ggcagcatca
     1321 acgaccggca gcccaccaag accaccccca gcatggccac caccacccag aggcccatca
     1381 ccgagagcga caaagaggac aacgaggaac ccatcgtgga cgacagcgac gaccagagcc
     1441 cttgcagcta cggcggcacc gtgttcacct acaccatcca cggcgacatc gacggcaacg
     1501 accacatgga aagcaacagc cacgacgagg acgactacat gcccgccgac gactacgact
     1561 tccaccccat ctacagctgg ctgggccggg acgacagctt ccaccagcgg cccagaaagc
     1621 cctaccacaa gcccacccgg aacacctggt acagcagata cgacaccttc ccctttggcg
     1681 cgccgtcgac ctccatcacg gcctataaga gtgaggggga gtcagcggag ttctccttcc
     1741 cactcaacct tggagaggaa agcctgcagg gagagttgag atggaaggca gagaaggctc
     1801 cttcttccca gtcctggatc accttctccc taaagaacca aaaggtgtct gtgcagaagt
     1861 ctactagcaa ccccaagttc cagctgtccg aaacgctccc actcaccctt cagatacccc
     1921 aggtctccct tcagtttgct ggttctggca acctgaccct gactctggac agagggatac
     1981 tgtatcagga agtgaacctg gtggtgatga aagtgactca gcccgacagc aacactttga
     2041 cctgtgaggt gatgggaccc acctcaccca agatgagact gatcttgaag caggagaatc
     2101 aggaggccag ggtctccagg caggagaaag tgattcaagt gcaggcccct gaagcagggg
     2161 tgtggcaatg tctactgagt gaaggtgaag aggtcaagat ggactccaag atccaggttt
     2221 tatccaaagg gttgaattcc ggcagcctgc accacatcct ggacgcccag aagatgctgt
     2281 ggaaccacag ggacaggaac ctgcctcctc tggcccctct gggcccccac caccatcacc
     2341 accattgagt ctagacacgt gattaattaa ggatcccccg acctcgacct ctggctaata
     2401 aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag
     2461 gacatatggg agggcaaatc atttggtcga gatccctcgg agatctctag ctagaggatc
     2521 gatccccgcc ccggacgaac taaacctgac tacgacatct ctgccccttc ttcgcggggc
     2581 agtgcatgta atcccttcag ttggttggta caacttgcca actgggccct gttccacatg
     2641 tgacacgggg ggggaccaaa cacaaagggg ttctctgact gtagttgaca tccttataaa
     2701 tggatgtgca catttgccaa cactgagtgg ctttcatcct ggagcagact ttgcagtctg
     2761 tggactgcaa cacaacattg cctttatgtg taactcttgg ctgaagctct tacaccaatg
     2821 ctgggggaca tgtacctccc aggggcccag gaagactacg ggaggctaca ccaacgtcaa
     2881 tcagaggggc ctgtgtagct accgataagc ggaccctcaa gagggcatta gcaatagtgt
     2941 ttataaggcc cccttgttaa ccctaaacgg gtagcatatg cttcccgggt agtagtatat
     3001 actatccaga ctaaccctaa ttcaatagca tatgttaccc aacgggaagc atatgctatc
     3061 gaattagggt tagtaaaagg gtcctaagga acagcgatat ctcccacccc atgagctgtc
     3121 acggttttat ttacatgggg tcaggattcc acgagggtag tgaaccattt tagtcacaag
     3181 ggcagtggct gaagatcaag gagcgggcag tgaactctcc tgaatcttcg cctgcttctt
     3241 cattctcctt cgtttagcta atagaataac tgctgagttg tgaacagtaa ggtgtatgtg
     3301 aggtgctcga aaacaaggtt tcaggtgacg cccccagaat aaaatttgga cggggggttc
     3361 agtggtggca ttgtgctatg acaccaatat aaccctcaca aaccccttgg gcaataaata
     3421 ctagtgtagg aatgaaacat tctgaatatc tttaacaata gaaatccatg gggtggggac
     3481 aagccgtaaa gactggatgt ccatctcaca cgaatttatg gctatgggca acacataatc
     3541 ctagtgcaat atgatactgg ggttattaag atgtgtccca ggcagggacc aagacaggtg
     3601 aaccatgttg ttacactcta tttgtaacaa ggggaaagag agtggacgcc gacagcagcg
     3661 gactccactg gttgtctcta acacccccga aaattaaacg gggctccacg ccaatggggc
     3721 ccataaacaa agacaagtgg ccactctttt ttttgaaatt gtggagtggg ggcacgcgtc
     3781 agcccccaca cgccgccctg cggttttgga ctgtaaaata agggtgtaat aacttggctg
     3841 attgtaaccc cgctaaccac tgcggtcaaa ccacttgccc acaaaaccac taatggcacc
     3901 ccggggaata cctgcataag taggtgggcg ggccaagata ggggcgcgat tgctgcgatc
     3961 tggaggacaa attacacaca cttgcgcctg agcgccaagc acagggttgt tggtcctcat
     4021 attcacgagg tcgctgagag cacggtgggc taatgttgcc atgggtagca tatactaccc
     4081 aaatatctgg atagcatatg ctatcctaat ctatatctgg gtagcatagg ctatcctaat
     4141 ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat
     4201 ttatatctgg gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat
     4261 ctatatctgg gtagtatatg ctatcctaat ctgtatccgg gtagcatatg ctatcctaat
     4321 agagattagg gtagtatatg ctatcctaat ttatatctgg gtagcatata ctacccaaat
     4381 atctggatag catatgctat cctaatctat atctgggtag catatgctat cctaatctat
     4441 atctgggtag cataggctat cctaatctat atctgggtag catatgctat cctaatctat
     4501 atctgggtag tatatgctat cctaatttat atctgggtag cataggctat cctaatctat
     4561 atctgggtag catatgctat cctaatctat atctgggtag tatatgctat cctaatctgt
     4621 atccgggtag catatgctat cctcatgata agctgtcaaa catgagaatt ttcttgaaga
     4681 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     4741 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     4801 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4861 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     4921 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     4981 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     5041 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     5101 tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac
     5161 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     5221 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     5281 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     5341 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     5401 gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc
     5461 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     5521 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     5581 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     5641 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     5701 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     5761 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     5821 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5881 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5941 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     6001 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     6061 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     6121 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     6181 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     6241 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     6301 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     6361 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     6421 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6481 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6541 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     6601 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     6661 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     6721 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     6781 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     6841 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     6901 catgattacg ccaagctcta gctagaggtc gaccaattct catgtttgac agcttatcat
     6961 cgcagatccg ggcaacgttg ttgccattgc tgcaggcgca gaactggtag gtatggaaga
     7021 tctatacatt gaatcaatat tggcaattag ccatattagt cattggttat atagcataaa
     7081 tcaatattgg ctattggcca ttgcatacgt tgtatctata tcataatatg tacatttata
     7141 ttggctcatg tccaatatga ccgc
//
