LOCUS       sequence-229137-        8916 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 8916)
  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 8916)
  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_127820-Cd4-bio-His
FEATURES             Location/Qualifiers
     source          1..8916
                     /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             4080..4097
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4149..4204
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4203..4222)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6440..6458)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6526..6630
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6631..6699,6700..7491)
                     /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(6849..6868)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7662..8250
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8151..8170
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8404..8421
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8538..8559
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8574..8579,8580..8597,8598..8604)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8612..8628
                     /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     8617..8639
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8636..8652
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8636..8652
                     /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 gcggccgccg
     1201 tgtcctacct gaagttcgag aagatcaacg gcgccagcga ggaaagcctg atcttcctga
     1261 acgacgccaa ccccaccgcc ctgctggacg ccagcgccaa cacccagcgg atcgtgtact
     1321 tcgcccagaa gtccccccca ctgggcctga agctgacctt cgcctgccac gtgaacaccc
     1381 aggccaagcc cctggacaag gtgtccctgt gctgccggtc cagcgtggaa gataccaagc
     1441 ggacctgcgt gcagcccgag gccgagggca ccttcagctg cctgaacacc gagttccact
     1501 tcagcgccga cgagacaacc ggcgccaact tcagcatcag cctgacccag gacggcctgc
     1561 tgagcaaccc catcagcggc ctgttcagct gcatcgtggg caccgacgag ctgaacgtga
     1621 tcgagagcaa cgagatcgag atccgggacg ccgtgttcta catcctgcag ctgaagggca
     1681 cccccagcgc caagatcgac gtggtgcccc ctatcagcga gttcccctac gagctgagat
     1741 gcgtggacga catcagcatc gtgtcccggt ggccctactg gatgcaccag ctggacggca
     1801 gatacggccc ctacagatgg gagtactgcc tgaccaccgg caagcccaac gccatcacct
     1861 gttacaagcg gcccatccag ttcagcccca acagcatcgt ggaattcatc aacggcagcg
     1921 tgtacctgac cgacgcccac ctggtggaaa gcaagtctat cgccatcgtg tgcaagcacg
     1981 tgggccagct gggccccatc cggaccttcg ccggcaaagt gaccagcacc gagagccccg
     2041 tggtgctgga aggcctgagc atcgacaaga ccaagaacag cagcagcacc aagctgtacc
     2101 ccctgaccac cctgagcagc agctacgtga tcaacaaggg cacaaagctg caggacttca
     2161 tgctgcgggc cctgtacaga cagccccagg gcgactacga gtttcactgg ttcaaggacg
     2221 agcagatgct gcacaaccgg atcgccacca agtacagcta caccctgccc agcctgatcg
     2281 acgagagcgt gtccggcgtg tacagcctgg tggtcaagtc caacgaggac atcagcgacc
     2341 ggctgatctt cacctacgac gtgaaggtgg tgtccccccc attcttcaag gacccccggt
     2401 gcctgaaaga actgttctac gtgatggaag gcaacaactt cggcaccgtg tgcgagttcg
     2461 acggccggtg gaacaccgtg gtgtacgtgg gcgtggccag cttcgaggcc cccacctacc
     2521 aggaactgcg gatgtccatc gataagaacc tgaagtacag caagcagcag ctgagccagc
     2581 tggacatcat ctttcacgtg gacggcgagg aaatgaacaa gcggatcacc gtgaacatca
     2641 ccaagctgaa gctgaaccag gactgcgagc tgagcatccg gctgcacagc acctacggcc
     2701 agagccggat cagcacctcc ctgaaagtgg tgcccaagcc ccagctgacc atcagcccca
     2761 gcgacgagaa ctgcatcaag gactgcgacg agacacccta caacgtgtcc tgcgccctga
     2821 acgagaacat cgtgcaggaa tggaagtcca agttcaacat cgagcccagc atcgactgga
     2881 tcatccagaa ccactggacc aaggcccacc tggaaagcac cggcctgggc cagttcatca
     2941 ccagctacgg cgagcacgac accaccctga ccgtgtggcc cgtgggcaag cccatcccct
     3001 tcatcgtgaa gcacgaagag agccacaaca agcagcccaa agaggccggc atcgaagaag
     3061 aggaagagga agtcatcgag aaagaaatca tgaacctgga caccgaggac gagcccgtgc
     3121 agatccccag cacaacaacc ctgaagcagt tcctggccca gaagatcggc ggcagcgagg
     3181 cccagccccc caaggatctg atcctgagct gctggatccg gctgagcgtg accggaaaag
     3241 aggccgagtt cgagatcccc aacggcctga tcagccacca cagcttcgcc agacaggccc
     3301 agttcgccca gagcagcgcc accaacagac tgatctacga cagcagatgg gagacagaca
     3361 ccgagctggc caagcagctg accgccttca gaaccttcac ccccgacatc atccccgcca
     3421 ccgcctctgg cgcgccgtcg acctccatca cggcctataa gagtgagggg gagtcagcgg
     3481 agttctcctt cccactcaac cttggagagg aaagcctgca gggagagttg agatggaagg
     3541 cagagaaggc tccttcttcc cagtcctgga tcaccttctc cctaaagaac caaaaggtgt
     3601 ctgtgcagaa gtctactagc aaccccaagt tccagctgtc cgaaacgctc ccactcaccc
     3661 ttcagatacc ccaggtctcc cttcagtttg ctggttctgg caacctgacc ctgactctgg
     3721 acagagggat actgtatcag gaagtgaacc tggtggtgat gaaagtgact cagcccgaca
     3781 gcaacacttt gacctgtgag gtgatgggac ccacctcacc caagatgaga ctgatcttga
     3841 agcaggagaa tcaggaggcc agggtctcca ggcaggagaa agtgattcaa gtgcaggccc
     3901 ctgaagcagg ggtgtggcaa tgtctactga gtgaaggtga agaggtcaag atggactcca
     3961 agatccaggt tttatccaaa gggttgaatt ccggcagcct gcaccacatc ctggacgccc
     4021 agaagatgct gtggaaccac agggacagga acctgcctcc tctggcccct ctgggccccc
     4081 accaccatca ccaccattga gtctagacac gtgattaatt aaggatcccc cgacctcgac
     4141 ctctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct
     4201 ctcactcgga aggacatatg ggagggcaaa tcatttggtc gagatccctc ggagatctct
     4261 agctagagga tcgatccccg ccccggacga actaaacctg actacgacat ctctgcccct
     4321 tcttcgcggg gcagtgcatg taatcccttc agttggttgg tacaacttgc caactgggcc
     4381 ctgttccaca tgtgacacgg ggggggacca aacacaaagg ggttctctga ctgtagttga
     4441 catccttata aatggatgtg cacatttgcc aacactgagt ggctttcatc ctggagcaga
     4501 ctttgcagtc tgtggactgc aacacaacat tgcctttatg tgtaactctt ggctgaagct
     4561 cttacaccaa tgctggggga catgtacctc ccaggggccc aggaagacta cgggaggcta
     4621 caccaacgtc aatcagaggg gcctgtgtag ctaccgataa gcggaccctc aagagggcat
     4681 tagcaatagt gtttataagg cccccttgtt aaccctaaac gggtagcata tgcttcccgg
     4741 gtagtagtat atactatcca gactaaccct aattcaatag catatgttac ccaacgggaa
     4801 gcatatgcta tcgaattagg gttagtaaaa gggtcctaag gaacagcgat atctcccacc
     4861 ccatgagctg tcacggtttt atttacatgg ggtcaggatt ccacgagggt agtgaaccat
     4921 tttagtcaca agggcagtgg ctgaagatca aggagcgggc agtgaactct cctgaatctt
     4981 cgcctgcttc ttcattctcc ttcgtttagc taatagaata actgctgagt tgtgaacagt
     5041 aaggtgtatg tgaggtgctc gaaaacaagg tttcaggtga cgcccccaga ataaaatttg
     5101 gacggggggt tcagtggtgg cattgtgcta tgacaccaat ataaccctca caaacccctt
     5161 gggcaataaa tactagtgta ggaatgaaac attctgaata tctttaacaa tagaaatcca
     5221 tggggtgggg acaagccgta aagactggat gtccatctca cacgaattta tggctatggg
     5281 caacacataa tcctagtgca atatgatact ggggttatta agatgtgtcc caggcaggga
     5341 ccaagacagg tgaaccatgt tgttacactc tatttgtaac aaggggaaag agagtggacg
     5401 ccgacagcag cggactccac tggttgtctc taacaccccc gaaaattaaa cggggctcca
     5461 cgccaatggg gcccataaac aaagacaagt ggccactctt ttttttgaaa ttgtggagtg
     5521 ggggcacgcg tcagccccca cacgccgccc tgcggttttg gactgtaaaa taagggtgta
     5581 ataacttggc tgattgtaac cccgctaacc actgcggtca aaccacttgc ccacaaaacc
     5641 actaatggca ccccggggaa tacctgcata agtaggtggg cgggccaaga taggggcgcg
     5701 attgctgcga tctggaggac aaattacaca cacttgcgcc tgagcgccaa gcacagggtt
     5761 gttggtcctc atattcacga ggtcgctgag agcacggtgg gctaatgttg ccatgggtag
     5821 catatactac ccaaatatct ggatagcata tgctatccta atctatatct gggtagcata
     5881 ggctatccta atctatatct gggtagcata tgctatccta atctatatct gggtagtata
     5941 tgctatccta atttatatct gggtagcata ggctatccta atctatatct gggtagcata
     6001 tgctatccta atctatatct gggtagtata tgctatccta atctgtatcc gggtagcata
     6061 tgctatccta atagagatta gggtagtata tgctatccta atttatatct gggtagcata
     6121 tactacccaa atatctggat agcatatgct atcctaatct atatctgggt agcatatgct
     6181 atcctaatct atatctgggt agcataggct atcctaatct atatctgggt agcatatgct
     6241 atcctaatct atatctgggt agtatatgct atcctaattt atatctgggt agcataggct
     6301 atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt agtatatgct
     6361 atcctaatct gtatccgggt agcatatgct atcctcatga taagctgtca aacatgagaa
     6421 ttttcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     6481 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     6541 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     6601 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     6661 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     6721 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     6781 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     6841 aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt
     6901 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     6961 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     7021 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     7081 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     7141 ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa
     7201 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     7261 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     7321 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     7381 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     7441 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     7501 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     7561 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     7621 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     7681 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     7741 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     7801 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     7861 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     7921 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     7981 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     8041 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     8101 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     8161 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     8221 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     8281 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     8341 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     8401 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     8461 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     8521 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     8581 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     8641 aacagctatg accatgatta cgccaagctc tagctagagg tcgaccaatt ctcatgtttg
     8701 acagcttatc atcgcagatc cgggcaacgt tgttgccatt gctgcaggcg cagaactggt
     8761 aggtatggaa gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt
     8821 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata
     8881 tgtacattta tattggctca tgtccaatat gaccgc
//
