LOCUS       sequence-393674-        2576 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pJL1 plasmid encoding superfolder GFP modified at the C-terminus 
            with 32 amino acids containing a non-permissible minimum optimal 
            O-linked recognition site (MOORmut) followed by a 6x-His tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2576)
  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 2576)
  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-MOORmut
FEATURES             Location/Qualifiers
     source          1..2576
                     /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             1505..1522
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1577..1595)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1591..1638
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1656..2471)
                     /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     2379..2398
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2463..2482
                     /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 gaattttacc gttcgccata acgttgaaga tggcagtgtg cagctggcag atcattatca
     1261 gcagaatacc ccgattggtg atggtccggt gctgctgccg gataatcatt atctgagcac
     1321 gcagaccgtt ctgtctaaag atccgaacga aaaaggcacg cgggaccaca tggttctgca
     1381 cgaatatgtg aatgcggcag gtattacggg ctcttctgga gggtctggcg atccacgcaa
     1441 tgtgggtggg gatttggact ggccggcggc agcgggtgca cctcaacccg gtaaacctcc
     1501 tcgtcatcac caccatcatc actaagtcga ccggctgcta acaaagcccg aaaggaagct
     1561 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg
     1621 gtcttgaggg gttttttgct gaaagccaat tctgattaga aaaactcatc gagcatcaaa
     1681 tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc
     1741 tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg
     1801 tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata
     1861 aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagc
     1921 ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca
     1981 ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga
     2041 tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc
     2101 agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt
     2161 ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg
     2221 atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca
     2281 tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca
     2341 tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca
     2401 tataaatcag catccatgtt ggaatttaat cgcggcttcg agcaagacgt ttcccgttga
     2461 atatggctca taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat
     2521 gatgatatat ttttatcttg tgcaatgtaa catcagagat tttgagacac aacgtg
//
