LOCUS       sequence-122687-        2486 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pJL1 expression vector with sfGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2486)
  AUTHORS   Martin RW, Des Soye BJ, Kwon YC, Kay J, Davis RG, Thomas PM, 
            Majewska NI, Chen CX, Marcum RD, Weiss MG, Stoddart AE, Amiram M, 
            Ranji Charna AK, Patel JR, Isaacs FJ, Kelleher NL, Hong SH, Jewett 
            MC
  TITLE     Cell-free protein synthesis from genomically recoded bacteria 
            enables multisite incorporation of noncanonical amino acids.
  JOURNAL   Nat Commun. 2018 Mar 23;9(1):1203. doi: 10.1038/s41467-018-03469-5.
  PUBMED    29572528
REFERENCE   2  (bases 1 to 2486)
  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: "2018-03-23"; volume: "9(1):1203. doi"; pages: " 
            10.1038/s41467-018-03469-5"
COMMENT     Sequence Label: pJL1
FEATURES             Location/Qualifiers
     source          1..2486
                     /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             1409..1432
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1487..1505)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1501..1548
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1566..2381)
                     /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     2289..2308
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2373..2392
                     /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 gtattacgtg gagccatccg cagttcgaaa aataagtcga
     1441 ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa
     1501 ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaagccaat
     1561 tctgattaga aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta
     1621 tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag
     1681 ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata
     1741 caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg
     1801 acgactgaat ccggtgagaa tggcaaaagc ttatgcattt ctttccagac ttgttcaaca
     1861 ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt
     1921 gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga
     1981 atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca
     2041 ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat
     2101 gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc
     2161 cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc
     2221 agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc
     2281 ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat
     2341 cgcggcttcg agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg
     2401 tttatgtaag cagacagttt tattgttcat gatgatatat ttttatcttg tgcaatgtaa
     2461 catcagagat tttgagacac aacgtg
//
