LOCUS       sequence-204384-        3006 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses sfGFP (a GFP) in Ecoli off a T7 promoter when pNP1 sfGFP 
            trigger is co-expressed.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3006)
  AUTHORS   Huang A, Nguyen PQ, Stark JC, Takahashi MK, Donghia N, Ferrante T, 
            Dy AJ, Hsu KJ, Dubner RS, Pardee K, Jewett MC, Collins JJ
  TITLE     BioBits Explorer: A modular synthetic biology education kit.
  JOURNAL   Sci Adv. 2018 Aug 1;4(8):eaat5105. doi: 10.1126/sciadv.aat5105. 
            eCollection 2018 Aug.
  PUBMED    30083608
REFERENCE   2  (bases 1 to 3006)
  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: "Sci Adv."; 
            date: "2018-08-1"; volume: "4(8):eaat5105. doi"; pages: " 
            10.1126/sciadv.aat5105. eCollection 2018 Aug"
COMMENT     Sequence Label: pNP1 sfGFP toehold switch
FEATURES             Location/Qualifiers
     source          1..3006
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     26..45
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        26..44
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             774..821
                     /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             822..845
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(984..1002)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      998..1045
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      complement(1263..1851)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1343..1362)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2023..2814,2815..2883))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2646..2665
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2884..2988)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gctagtggtg ctagccccgc gaaattaata cgactcacta tagggatcac ttattgtcgt
       61 ctttgtatgt ctgtaaacag aggagataca gaatgacaaa gacgaacctg gcggcagcgc
      121 aaaagatgag caaaggtgaa gaactgttta ccggcgttgt gccgattctg gtggaactgg
      181 atggcgatgt gaacggtcac aaattcagcg tgcgtggtga aggtgaaggc gatgccacga
      241 ttggcaaact gacgctgaaa tttatctgca ccaccggcaa actgccggtg ccgtggccga
      301 cgctggtgac caccctgacc tatggcgttc agtgttttag tcgctatccg gatcacatga
      361 aacgtcacga tttctttaaa tctgcaatgc cggaaggcta tgtgcaggaa cgtacgatta
      421 gctttaaaga tgatggcaaa tataaaacgc gcgccgttgt gaaatttgaa ggcgataccc
      481 tggtgaaccg cattgaactg aaaggcacgg attttaaaga agatggcaat atcctgggcc
      541 ataaactgga atacaacttt aatagccata atgtttatat tacggcggat aaacagaaaa
      601 atggcatcaa agcgaatttt accgttcgcc ataacgttga agatggcagt gtgcagctgg
      661 cagatcatta tcagcagaat accccgattg gtgatggtcc ggtgctgctg ccggataatc
      721 attatctgag cacgcagacc gttctgtcta aagatccgaa cgaaaaaggc acgcgggacc
      781 acatggttct gcacgaatat gtgaatgcgg caggtattac gtggagccat ccgcagttcg
      841 aaaaataaag gatcccggga attctcgagt aaggttaacc tgcaggaggc ctttaattaa
      901 ggtggtgcgg ccgcgctagc ggtcccgggg gatcgatccg gctgctaaca aagcccgaaa
      961 ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc
     1021 taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggaa gcttggcact
     1081 ggccgaccgg ggtcgagcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     1141 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     1201 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     1261 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     1321 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     1381 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     1441 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     1501 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     1561 aactatcgtc ttgagtccaa cccgctaaga cacgacttat cgccactggc agcagccact
     1621 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     1681 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     1741 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     1801 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     1861 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacagat ccgggatttt
     1921 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     1981 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     2041 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     2101 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     2161 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     2221 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     2281 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     2341 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     2401 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     2461 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     2521 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     2581 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     2641 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     2701 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     2761 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     2821 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     2881 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     2941 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     3001 aaaagt
//
