LOCUS       sequence-331133-        2612 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  malachite green aptamer tagged sfGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2612)
  TITLE     Sensitizing cell-free molecular detection using cascaded genetic 
            circuitry
REFERENCE   2  (bases 1 to 2612)
  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"
COMMENT     Sequence Label: pJBL7070
FEATURES             Location/Qualifiers
     source          1..2612
                     /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             684..704
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             join(714..716,717..1430)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             714..1427
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(744..772)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1367..1387
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1613..1631)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1627..1674
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1692..2507)
                     /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     2415..2434
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2499..2518
                     /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 ggaggaagga tctatgagca
      721 aaggagaaga acttttcact ggagttgtcc caattcttgt tgaattagat ggtgatgtta
      781 atgggcacaa attttctgtc cgtggagagg gtgaaggtga tgctacaaac ggaaaactca
      841 cccttaaatt tatttgcact actggaaaac tacctgttcc gtggccaaca cttgtcacta
      901 ctctgaccta tggtgttcaa tgcttttccc gttatccgga tcacatgaaa cggcatgact
      961 ttttcaagag tgccatgccc gaaggttatg tacaggaacg cactatatct ttcaaagatg
     1021 acgggaccta caagacgcgt gctgaagtca agtttgaagg tgataccctt gttaatcgta
     1081 tcgagttaaa gggtattgat tttaaagaag atggaaacat tcttggacac aaactcgagt
     1141 acaactttaa ctcacacaat gtatacatca cggcagacaa acaaaagaat ggaatcaaag
     1201 ctaacttcaa aattcgccac aacgttgaag atggttccgt tcaactagca gaccattatc
     1261 aacaaaatac tccaattggc gatggccctg tccttttacc agacaaccat tacctgtcga
     1321 cacaatctgt cctttcgaaa gatcccaacg aaaagcgtga ccacatggtc cttcttgagt
     1381 ttgtaactgc tgctgggatt acacatggca tggatgagct ctacaaataa ggatctgaag
     1441 gggatcccga ctggcgagag ccaggtaacg aatggatcgg gtcggcatgg catctccacc
     1501 tcctcgcggt ccgacctggg catccgaagg aggacgtcgt ccactcggat ggctaaggga
     1561 ggtcgaccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag
     1621 caataactag cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa
     1681 gccaattctg attagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca
     1741 ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg
     1801 aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca
     1861 tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca
     1921 tgagtgacga ctgaatccgg tgagaatggc aaaagcttat gcatttcttt ccagacttgt
     1981 tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc
     2041 attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg acaattacaa
     2101 acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct
     2161 gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc agtggtgagt
     2221 aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc
     2281 gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca
     2341 tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct
     2401 gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa
     2461 tttaatcgcg gcttcgagca agacgtttcc cgttgaatat ggctcataac accccttgta
     2521 ttactgttta tgtaagcaga cagttttatt gttcatgatg atatattttt atcttgtgca
     2581 atgtaacatc agagattttg agacacaacg tg
//
