LOCUS       sequence-413051-        5248 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses fucosidase E1_10125 from Ruminococcus gnavus in BL21 
            cells, purified using His-tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5248)
  AUTHORS   Spruit C, Palme DI, Li T, Rios Carrasco M, Gabarroca Garcia A, Sweet
            I, Kuryshko M, Maliepaard JCL, Reiding KR, Scheibner D, Boons G-J, 
            Abdelwhab EM, de Vries RP
  TITLE     H7 influenza A viruses bind sialyl-LewisX, a potential intermediate 
            receptor between species
  JOURNAL   bioRxiv 2023.12.15.571923
REFERENCE   2  (bases 1 to 5248)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2023.12.15.571923"
COMMENT     Sequence Label: pET23A-Fucosidase-E1_10125-Ruminococcus_gnavus...
FEATURES             Location/Qualifiers
     source          1..5248
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      12..467
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(99..118)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     309..330
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        493..597
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(598..666,667..1458)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(816..835)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1629..2217
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2118..2137
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2371..2388
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2403..2542
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2468..2487)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2628..2650
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2644..2835)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     3344..3363
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3344..3362
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             3394..3416
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4190..4201
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             5086..5109
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(5162..5180)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5176..5223
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
       61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
      121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
      241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
      361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
      421 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
      481 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
      541 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
      601 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
      661 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
      721 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
      781 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
      841 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
      901 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
      961 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     1021 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     1081 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     1141 gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     1201 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     1261 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     1321 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     1381 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     1441 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     1501 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     1561 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     1621 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     1681 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     1741 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     1801 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     1861 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     1921 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     1981 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     2041 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     2101 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     2161 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     2221 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     2281 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     2341 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     2401 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg caatggtgca
     2461 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct
     2521 acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
     2581 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     2641 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc
     2701 agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag
     2761 tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt
     2821 ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat
     2881 accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt
     2941 actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat
     3001 cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca
     3061 gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc
     3121 cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt
     3181 tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt
     3241 aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca tgcgcacccg
     3301 tggccaggac ccaacgctgc ccgagatctc gatcccgcga aattaatacg actcactata
     3361 gggagaccac aacggtttcc ctctagaaat aattttgttt aactttaaga aggagatata
     3421 catatggaaa caacaaattg gattggggat gaaaatttaa caggaaatgc ggaagctcca
     3481 gcaaaagatg atgtagtgcc ggataagaac cagttccgtt atcagaaaga ggagctggca
     3541 gcattttgtc attttggacc gaatacattc aatgagattg aatggggaga gcactatggg
     3601 aatcagaagc caagtgagat ttttacattg aaaaatgatt ttgatgcgga gactttagta
     3661 aaaacattaa aagatgctgg ttttaaaaag ctgattgtga ctgcaaagca tcatgatgga
     3721 ttctgtattt gggatagtga acacacagaa tatgatgtaa aagcatctgg ctataagaat
     3781 aaaaacggag aatcggatat tttggcagag atatcaaaag catgtaccga tcagaatatg
     3841 gacatgggat tgtatttgtc tccgtgggat attcatgagc caagttatgg ttataaagat
     3901 gaacatggaa atccaacaac tccagacaaa gatgcaaagg attataatga gttttacaat
     3961 aatcaactgg aagagattct tggaaatcca aagtatggaa atgatggtca ttttgttgag
     4021 gtatggatgg atggtgccaa aggaagtggc gctaatgcac aggaatacga ctttaaaaaa
     4081 tggtttaaga caattcagga taacgaggga aaagcagcgg gatatgatgc agactgtatg
     4141 ttatttgggg cagaagcata taccacagta cgctggattg gaaatgaatt gggaattgca
     4201 ggaaaggata cgtggtcaaa atctaaagta gacaaggata agaacacaat taatagtaac
     4261 aagcagggaa atgcgacggt tggttttgaa gatggagatc agtggacagt tccggaggca
     4321 gatgcgcgta ttacttctgg ttggttctgg ggaactaaga agaatactcc aaaaacgatg
     4381 gaagaactga gtgatatgta ttttaattca gttggacata atgcaacatt gctactgaat
     4441 gtacctccga acaatcaggg aacagtagac aaagcaattc tggatcgtgt gacagaattt
     4501 ggaaataata tcaaagcaac atttaaaact aatctggcaa aagcagaggg ggcttctgta
     4561 aaagtttctg aagtcagagg tggtgcaaaa gaatataaac caggcaatat gattgacgat
     4621 aatgatgaga catattgggc tacaagtgat ggcaaaaaat ccggtgaaat tctgattgat
     4681 ttaggaaaag aaacaaagtt tgatgttgta tcaattgagg aggcaattca gaacggtcaa
     4741 agaattaata attacaaagt agagtatcga aatggtgata gtggtacatg gactctgttg
     4801 gaagaaggaa aaactattgg tgcgaaacgc ctttgcagaa caagtgagac aaccgcacgt
     4861 cagattaaaa ttacagtggg tacgtgtgat ggaaaagttc ctatgatcag tgaaattgga
     4921 gtatacaagt ctactgagga tatggaaaaa ccaaatccga ttccaaaggg aatggaagtc
     4981 attgatgtgg aagataagga tgtagcagat ggaaaaggat ttacgttcaa aggaaaatgg
     5041 aatccagaaa atcagccgca gtatatcaat ggtgtcgacc tcgagcacca ccaccaccac
     5101 caccaccact gagatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
     5161 accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     5221 ttgctgaaag gaggaactat atccggat
//
