LOCUS       sequence-234412-        7500 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 7500)
  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 7500)
  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_180600-Cd4-bio-His
FEATURES             Location/Qualifiers
     source          1..7500
                     /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             2278..2295
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2347..2402
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2401..2420)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4638..4656)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4724..4828
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4829..4897,4898..5689)
                     /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(5047..5066)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5860..6448
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6349..6368
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6602..6619
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6736..6757
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6772..6777,6778..6795,6796..6802)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6810..6826
                     /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     6815..6837
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6834..6850
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6834..6850
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7121..7500
                     /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 gccagccagt acgacaacga cgatcaccac
      841 cactactacg agcccagcaa gagcctgcag agccccacca tcctgaagca gatcgaggac
      901 gagtgcaaca agtgcgaccg gaagaagtgc agccccatca tccagtgcaa cgtgggcatc
      961 gtgcgggaca agtgcggctg ctgcgacatc tgcgccctgg aagaggccca gctgtgcaac
     1021 atccccaacg acttcatcaa cggcattctg aagcagggca tcacttggta cggcatctgc
     1081 ggcaccgacc tggaatgccg gaagcggacc gacatcgaca gcaagatcat cgagagccag
     1141 agcatctgct actgcgtgaa gtccggcaaa gtgtgcggca gcgacggcat cacctacagc
     1201 aagtgcaaga tgaacgccac catcatcagc agcaacggca ccgtgatccc catcagcgag
     1261 ggcccctgtc agaccgtgcc cagcgtgaag ctgaccgtgc tgaaccagac cgtgatcgag
     1321 ggcaccaacg tgaccctgat ctgcgaggcc agaggctacc ccatccccac catcaactgg
     1381 tacttcaaca agcccaacca gaagcaggac aagatccaga tgcccggcaa ctacaaggac
     1441 gtgaccgtgt ccgtgcgggg aggcaccgag gaaacccaca ccatcagcta cctgcagatc
     1501 accagcttca ccctggaata cgagggcgag tacatctgca tggccgacaa catcgtgggc
     1561 atcaaggccc agggcaccag cctcgtgcgg aacgccgcca gcagcatcta cagcagcagc
     1621 agctctggcg cgccgtcgac ctccatcacg gcctataaga gtgaggggga gtcagcggag
     1681 ttctccttcc cactcaacct tggagaggaa agcctgcagg gagagttgag atggaaggca
     1741 gagaaggctc cttcttccca gtcctggatc accttctccc taaagaacca aaaggtgtct
     1801 gtgcagaagt ctactagcaa ccccaagttc cagctgtccg aaacgctccc actcaccctt
     1861 cagatacccc aggtctccct tcagtttgct ggttctggca acctgaccct gactctggac
     1921 agagggatac tgtatcagga agtgaacctg gtggtgatga aagtgactca gcccgacagc
     1981 aacactttga cctgtgaggt gatgggaccc acctcaccca agatgagact gatcttgaag
     2041 caggagaatc aggaggccag ggtctccagg caggagaaag tgattcaagt gcaggcccct
     2101 gaagcagggg tgtggcaatg tctactgagt gaaggtgaag aggtcaagat ggactccaag
     2161 atccaggttt tatccaaagg gttgaattcc ggcagcctgc accacatcct ggacgcccag
     2221 aagatgctgt ggaaccacag ggacaggaac ctgcctcctc tggcccctct gggcccccac
     2281 caccatcacc accattgagt ctagacacgt gattaattaa ggatcccccg acctcgacct
     2341 ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct
     2401 cactcggaag gacatatggg agggcaaatc atttggtcga gatccctcgg agatctctag
     2461 ctagaggatc gatccccgcc ccggacgaac taaacctgac tacgacatct ctgccccttc
     2521 ttcgcggggc agtgcatgta atcccttcag ttggttggta caacttgcca actgggccct
     2581 gttccacatg tgacacgggg ggggaccaaa cacaaagggg ttctctgact gtagttgaca
     2641 tccttataaa tggatgtgca catttgccaa cactgagtgg ctttcatcct ggagcagact
     2701 ttgcagtctg tggactgcaa cacaacattg cctttatgtg taactcttgg ctgaagctct
     2761 tacaccaatg ctgggggaca tgtacctccc aggggcccag gaagactacg ggaggctaca
     2821 ccaacgtcaa tcagaggggc ctgtgtagct accgataagc ggaccctcaa gagggcatta
     2881 gcaatagtgt ttataaggcc cccttgttaa ccctaaacgg gtagcatatg cttcccgggt
     2941 agtagtatat actatccaga ctaaccctaa ttcaatagca tatgttaccc aacgggaagc
     3001 atatgctatc gaattagggt tagtaaaagg gtcctaagga acagcgatat ctcccacccc
     3061 atgagctgtc acggttttat ttacatgggg tcaggattcc acgagggtag tgaaccattt
     3121 tagtcacaag ggcagtggct gaagatcaag gagcgggcag tgaactctcc tgaatcttcg
     3181 cctgcttctt cattctcctt cgtttagcta atagaataac tgctgagttg tgaacagtaa
     3241 ggtgtatgtg aggtgctcga aaacaaggtt tcaggtgacg cccccagaat aaaatttgga
     3301 cggggggttc agtggtggca ttgtgctatg acaccaatat aaccctcaca aaccccttgg
     3361 gcaataaata ctagtgtagg aatgaaacat tctgaatatc tttaacaata gaaatccatg
     3421 gggtggggac aagccgtaaa gactggatgt ccatctcaca cgaatttatg gctatgggca
     3481 acacataatc ctagtgcaat atgatactgg ggttattaag atgtgtccca ggcagggacc
     3541 aagacaggtg aaccatgttg ttacactcta tttgtaacaa ggggaaagag agtggacgcc
     3601 gacagcagcg gactccactg gttgtctcta acacccccga aaattaaacg gggctccacg
     3661 ccaatggggc ccataaacaa agacaagtgg ccactctttt ttttgaaatt gtggagtggg
     3721 ggcacgcgtc agcccccaca cgccgccctg cggttttgga ctgtaaaata agggtgtaat
     3781 aacttggctg attgtaaccc cgctaaccac tgcggtcaaa ccacttgccc acaaaaccac
     3841 taatggcacc ccggggaata cctgcataag taggtgggcg ggccaagata ggggcgcgat
     3901 tgctgcgatc tggaggacaa attacacaca cttgcgcctg agcgccaagc acagggttgt
     3961 tggtcctcat attcacgagg tcgctgagag cacggtgggc taatgttgcc atgggtagca
     4021 tatactaccc aaatatctgg atagcatatg ctatcctaat ctatatctgg gtagcatagg
     4081 ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagtatatg
     4141 ctatcctaat ttatatctgg gtagcatagg ctatcctaat ctatatctgg gtagcatatg
     4201 ctatcctaat ctatatctgg gtagtatatg ctatcctaat ctgtatccgg gtagcatatg
     4261 ctatcctaat agagattagg gtagtatatg ctatcctaat ttatatctgg gtagcatata
     4321 ctacccaaat atctggatag catatgctat cctaatctat atctgggtag catatgctat
     4381 cctaatctat atctgggtag cataggctat cctaatctat atctgggtag catatgctat
     4441 cctaatctat atctgggtag tatatgctat cctaatttat atctgggtag cataggctat
     4501 cctaatctat atctgggtag catatgctat cctaatctat atctgggtag tatatgctat
     4561 cctaatctgt atccgggtag catatgctat cctcatgata agctgtcaaa catgagaatt
     4621 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     4681 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     4741 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4801 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     4861 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     4921 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     4981 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     5041 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
     5101 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     5161 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     5221 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     5281 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     5341 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact
     5401 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     5461 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     5521 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     5581 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     5641 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     5701 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     5761 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     5821 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     5881 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     5941 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     6001 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     6061 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     6121 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     6181 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     6241 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     6301 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     6361 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     6421 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     6481 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     6541 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     6601 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     6661 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     6721 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     6781 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     6841 cagctatgac catgattacg ccaagctcta gctagaggtc gaccaattct catgtttgac
     6901 agcttatcat cgcagatccg ggcaacgttg ttgccattgc tgcaggcgca gaactggtag
     6961 gtatggaaga tctatacatt gaatcaatat tggcaattag ccatattagt cattggttat
     7021 atagcataaa tcaatattgg ctattggcca ttgcatacgt tgtatctata tcataatatg
     7081 tacatttata ttggctcatg tccaatatga ccgccatgtt gacattgatt attgactagt
     7141 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     7201 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     7261 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     7321 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     7381 ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     7441 accttacggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
//
