LOCUS       sequence-392565-        6169 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pSN18 derivative encoding C.jejuni PglB with C-terminal FLAG epitope
            tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6169)
  AUTHORS   Hershewe JM, Warfel KF, Iyer SM, Peruzzi JA, Sullivan CJ, Roth EW, 
            DeLisa MP, Kamat NP, Jewett MC
  TITLE     Improving cell-free glycoprotein synthesis by characterizing and 
            enriching native membrane vesicles.
  JOURNAL   Nat Commun. 2021 Apr 22;12(1):2363. doi: 10.1038/s41467-021-22329-3.
  PUBMED    33888690
REFERENCE   2  (bases 1 to 6169)
  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: "Nat 
            Commun."; date: "2021-04-22"; volume: "12(1):2363. doi"; pages: " 
            10.1038/s41467-021-22329-3"
COMMENT     Sequence Label: pSF-CjPglB
FEATURES             Location/Qualifiers
     source          1..6169
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2243..2266
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2331..2348)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2501..2587
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2679..2706
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2725..2816
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2817..2885,2886..3677)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3035..3054)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3848..4436
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4337..4356
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4590..4607
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4622..4762
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4688..4707)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4848..4870
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5026..5904)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        join(5931..6169,1..46)
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     6138..6157
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
ORIGIN
        1 gatcctacct gacgcttttt atcgcaactc tctactgttt ctccataccc gttttttggg
       61 ctaacaggag gaattaacca tggcatctag tatgttgaaa aaagagtatt taaaaaaccc
      121 ttatttagtt ttgtttgcga tgattatatt agcttatgtt tttagtgtat tttgcaggtt
      181 ttattgggtt tggtgggcaa gtgagtttaa tgagtatttt ttcaataatc agttaatgat
      241 catttcaaat gatggctatg cttttgctga gggcgcaaga gatatgatag caggttttca
      301 tcagcctaat gatttgagtt attatggatc ttctttatcc gcgcttactt attggcttta
      361 taaaatcaca cctttttctt ttgaaagtat cattttatat atgagtactt ttttatcttc
      421 tttggtggtg attcctacta ttttgctagc taacgaatac aaacgtcctt taatgggctt
      481 tgtagctgct cttttagcaa gtatagcaaa cagttattat aatcgcacta tgagtgggta
      541 ttatgatacg gatatgctgg taattgtttt gcctatgttt attttatttt ttatggtaag
      601 aatgatttta aaaaaagact ttttttcatt gattgccttg ccgttattta taggaattta
      661 tctttggtgg tatccttcaa gttatacttt aaatgtagct ttaattggac tttttttaat
      721 ttatacactt atttttcata gaaaagaaaa gattttttat atagctgtga ttttgtcttc
      781 tcttactctt tcaaatatag catggtttta tcaaagtgcc attatagtaa tactttttgc
      841 tttattcgcc ttagagcaaa aacgcttaaa ttttatgatt ataggaattt taggtagtgc
      901 aactttgata tttttgattt taagtggtgg ggttgatcct atactttatc agcttaaatt
      961 ttatattttt agaagtgatg aaagtgcgaa tttaacgcag ggctttatgt attttaatgt
     1021 caatcaaacc atacaagaag ttgaaaatgt agatcttagc gaatttatgc gaagaattag
     1081 tggtagtgaa attgtttttt tgttttcttt gtttggtttt gtatggcttt tgagaaaaca
     1141 taaaagtatg attatggctt tacctatatt ggtgcttggg tttttagcct taaaaggggg
     1201 gcttagattt accatttatt ctgtacctgt aatggcctta ggatttggtt ttttattgag
     1261 cgagtttaag gctataatgg ttaaaaaata tagccaatta acttcaaatg tttgtattgt
     1321 ttttgcaact attttgactt tagctccagt atttatccat atttacaact ataaagcgcc
     1381 aacagttttt tctcaaaatg aagcatcatt attaaatcaa ttaaaaaata tagccaatag
     1441 agaagattat gtggtaactt ggtgggatta tggttatcct gtgcgttatt atagcgatgt
     1501 gaaaacttta gtagatggtg gaaagcattt aggtaaggat aattttttcc cttcttttgc
     1561 tttaagcaaa gatgaacaag ctgcagctaa tatggcaaga cttagtgtag aatatacaga
     1621 aaaaagcttt tatgctccgc aaaatgatat tttaaaaaca gacattttgc aagccatgat
     1681 gaaagattat aatcaaagca atgtggattt gtttctagct tcattatcaa aacctgattt
     1741 taaaatcgat acgccaaaaa ctcgtgatat ttatctttat atgcccgcta gaatgtcttt
     1801 gattttttct acggtggcta gtttttcttt tattaattta gatacaggag ttttggataa
     1861 accttttacc tttagcacag cttatccact tgatgttaaa aatggagaaa tttatcttag
     1921 caacggagtg gttttaagcg atgattttag aagttttaaa ataggtgata atgtggtttc
     1981 tgtaaatagt atcgtagaga ttaattctat taaacaaggt gaatacaaaa tcactccaat
     2041 tgatgataag gctcagtttt atatttttta tttaaaggat agtgctattc cttacgcaca
     2101 atttatttta atggataaaa ccatgtttaa tagtgcttat gtgcaaatgt tttttttagg
     2161 aaattatgat aagaatttat ttgacttggt gattaattct agagatgcta aggtttttaa
     2221 acttaaaatt cctgcaggag cagactacaa ggacgatgac gacaagggat aagtcgacgt
     2281 ttaaacggtc tccagcttgg ctgttttggc ggatgagaga agattttcag cctgatacag
     2341 attaaatcag aacgcagaag cggtctgata aaacagaatt tgcctggcgg cagtagcgcg
     2401 gtggtcccac ctgaccccat gccgaactca gaagtgaaac gccgtagcgc cgatggtagt
     2461 gtggggtctc cccatgcgag agtagggaac tgccaggcat caaataaaac gaaaggctca
     2521 gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag
     2581 gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc
     2641 aggacgcccg ccataaactg ccaggcatca aattaagcag aaggccatcc tgacggatgg
     2701 cctttttgcg tttctacaaa ctcttttgtt tatttttcta aatacattca aatatgtatc
     2761 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     2821 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     2881 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     2941 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     3001 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg
     3061 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     3121 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     3181 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     3241 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     3301 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     3361 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     3421 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     3481 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     3541 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     3601 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     3661 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     3721 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     3781 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     3841 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     3901 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     3961 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     4021 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4081 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4141 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4201 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4261 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4321 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4381 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4441 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     4501 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     4561 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     4621 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca
     4681 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct
     4741 acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
     4801 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     4861 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagcagatca attcgcgcgc
     4921 gaaggcgaag cggcatgcat aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac
     4981 cctatgctac tccgtcaagc cgtcaattgt ctgattcgtt accaattatg acaacttgac
     5041 ggctacatca ttcacttttt cttcacaacc ggcacggaac tcgctcgggc tggccccggt
     5101 gcatttttta aatacccgcg agaaatagag ttgatcgtca aaaccaacat tgcgaccgac
     5161 ggtggcgata ggcatccggg tggtgctcaa aagcagcttc gcctggctga tacgttggtc
     5221 ctcgcgccag cttaagacgc taatccctaa ctgctggcgg aaaagatgtg acagacgcga
     5281 cggcgacaag caaacatgct gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg
     5341 atcgctgatg tactgacaag cctcgcgtac ccgattatcc atcggtggat ggagcgactc
     5401 gttaatcgct tccatgcgcc gcagtaacaa ttgctcaagc agatttatcg ccagcagctc
     5461 cgaatagcgc ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg
     5521 cggctggtgc gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg acggccagtt
     5581 aagccattca tgccagtagg cgcgcggacg aaagtaaacc cactggtgat accattcgcg
     5641 agcctccgga tgacgaccgt agtgatgaat ctctcctggc gggaacagca aaatatcacc
     5701 cggtcggcaa acaaattctc gtccctgatt tttcaccacc ccctgaccgc gaatggtgag
     5761 attgagaata taacctttca ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt
     5821 ggcctcaatc ggcgttaaac ccgccaccag atgggcatta aacgagtatc ccggcagcag
     5881 gggatcattt tgcgcttcag ccatactttt catactcccg ccattcagag aagaaaccaa
     5941 ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct tctcgctaac
     6001 caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca aagccatgac
     6061 aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg attatttgca
     6121 cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcg
//
