LOCUS       sequence-393679-        2570 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pJL1 plasmid encoding superfolder GFP modified at the C-terminus 
            with 30 amino acids containing a non-permissible AQNAT sequon 
            followed by a 6x-His tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2570)
  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 2570)
  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: pJL1-sfGFP-AQNAT
FEATURES             Location/Qualifiers
     source          1..2570
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      17..605
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     506..525
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     633..652
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        633..651
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             683..705
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1361..1408
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
     CDS             1424..1453
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1499..1516
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1571..1589)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1585..1632
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1650..2465)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2373..2392
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2457..2476
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
       61 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
      121 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
      181 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
      241 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
      301 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      361 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      421 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      481 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      541 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      601 ggaaaaacgc cagcaacgcg atcccgcgaa attaatacga ctcactatag ggagaccaca
      661 acggtttccc tctagaaata attttgttta actttaagaa ggagatatac atatgagcaa
      721 aggtgaagaa ctgtttaccg gcgttgtgcc gattctggtg gaactggatg gcgatgtgaa
      781 cggtcacaaa ttcagcgtgc gtggtgaagg tgaaggcgat gccacgattg gcaaactgac
      841 gctgaaattt atctgcacca ccggcaaact gccggtgccg tggccgacgc tggtgaccac
      901 cctgacctat ggcgttcagt gttttagtcg ctatccggat cacatgaaac gtcacgattt
      961 ctttaaatct gcaatgccgg aaggctatgt gcaggaacgt acgattagct ttaaagatga
     1021 tggcaaatat aaaacgcgcg ccgttgtgaa atttgaaggc gataccctgg tgaaccgcat
     1081 tgaactgaaa ggcacggatt ttaaagaaga tggcaatatc ctgggccata aactggaata
     1141 caactttaat agccataatg tttatattac ggcggataaa cagaaaaatg gcatcaaagc
     1201 gcagtttacc gttcgccata acgttgaaga tggcagtgtg cagctggcag atcattatca
     1261 gcagaatacc ccgattggtg atggtccggt gctgctgccg gataatcatt atctgagcac
     1321 gcagaccgtt ctgtctaaag atccgaacga aaaaggcacg cgggaccaca tggttctgca
     1381 cgaatatgtg aatgcggcag gtattacgct aggtgcggcc gcagaacaaa aactcatctc
     1441 agaagaggat ctgaatgggg ccgcactcga gggtggcgct cagaacgcga ccggcggtca
     1501 tcaccatcat caccattaag tcgaccggct gctaacaaag cccgaaagga agctgagttg
     1561 gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg
     1621 aggggttttt tgctgaaagc caattctgat tagaaaaact catcgagcat caaatgaaac
     1681 tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat
     1741 gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg
     1801 attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta
     1861 tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa aagcttatgc
     1921 atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca
     1981 tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg
     2041 ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca
     2101 tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg
     2161 gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc
     2221 ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg
     2281 gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat
     2341 cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa
     2401 tcagcatcca tgttggaatt taatcgcggc ttcgagcaag acgtttcccg ttgaatatgg
     2461 ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt tcatgatgat
     2521 atatttttat cttgtgcaat gtaacatcag agattttgag acacaacgtg
//
