LOCUS       sequence-320898-        2499 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reporter plasmid for T7-driven E. coli cell-free protein synthesis 
            with detection via HiBiT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2499)
  AUTHORS   Dudley QM, Cai YM, Kallam K, Debreyne H, Carrasco Lopez JA, Patron 
            NJ
  TITLE     Biofoundry-assisted expression and characterization of plant 
            proteins.
  JOURNAL   Synth Biol (Oxf). 2021 Sep 11;6(1):ysab029. doi: 
            10.1093/synbio/ysab029. eCollection 2021.
  PUBMED    34693026
REFERENCE   2  (bases 1 to 2499)
  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: "Synth Biol 
            (Oxf)."; date: "2021-09-11"; volume: "6(1):ysab029. doi"; pages: " 
            10.1093/synbio/ysab029. eCollection 2021"
COMMENT     Sequence Label: pEPQDKN0248
FEATURES             Location/Qualifiers
     source          1..2499
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             570..617
                     /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"
     primer_bind     complement(709..727)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      723..770
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(788..1603)
                     /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     1511..1530
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1595..1614
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1725..2313
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2214..2233
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2341..2360
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2341..2359
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2391..2413
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 tcagcgtgcg tggtgaaggt gaaggcgatg ccacgattgg caaactgacg ctgaaattta
       61 tctgcaccac cggcaaactg ccggtgccgt ggccgacgct ggtgaccacc ctgacctatg
      121 gcgttcagtg ttttagtcgc tatccggatc acatgaaacg tcacgatttc tttaaatctg
      181 caatgccgga aggctatgtg caggaacgta cgattagctt taaagatgat ggcaaatata
      241 aaacgcgcgc cgttgtgaaa tttgaaggcg ataccctggt gaaccgcatt gaactgaaag
      301 gcacggattt taaagaagat ggcaatatcc tgggccataa actggaatac aactttaata
      361 gccataatgt ttatattacg gcggataaac agaaaaatgg catcaaagcg aattttaccg
      421 ttcgccataa cgttgaagat ggcagtgtgc agctggcaga tcattatcag cagaataccc
      481 cgattggtga tggtccggtg ctgctgccgg ataatcatta tctgagcacg cagaccgttc
      541 tgtctaaaga tccgaacgaa aaaggcacgc gggaccacat ggttctgcac gaatatgtga
      601 atgcggcagg tattacgggt tcggtgagcg gctggcggct gttcaagaag attagctgag
      661 cttcggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
      721 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaagcca
      781 attctgatta gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat
      841 tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc
      901 agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa
      961 tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag
     1021 tgacgactga atccggtgag aatggcaaaa gcttatgcat ttctttccag acttgttcaa
     1081 caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc
     1141 gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag
     1201 gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt tcacctgaat
     1261 caggatattc ttctaatacc tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc
     1321 atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca
     1381 gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt
     1441 tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc gcacctgatt
     1501 gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta
     1561 atcgcggctt cgagcaagac gtttcccgtt gaatatggct cataacaccc cttgtattac
     1621 tgtttatgta agcagacagt tttattgttc atgatgatat atttttatct tgtgcaatgt
     1681 aacatcagag attttgagac acaacgtgag atcaaaggat cttcttgaga tccttttttt
     1741 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     1801 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     1861 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     1921 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     1981 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     2041 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     2101 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     2161 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     2221 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     2281 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcgat cccgcgaaat
     2341 taatacgact cactataggg acaccacaac ggtttccctc tagaaataat tttgtttaac
     2401 tttaagaagg agatatacaa atgagcaaag gtgaagaact gtttaccggc gttgtgccga
     2461 ttctggtgga actggatggc gatgtgaacg gtcacaaat
//
