LOCUS       sequence-209404-        3079 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses sfGFP (a GFP) in Ecoli off a T7 promoter when pNP1 OR 
            input A and pNP1 OR input B are co-expressed.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3079)
  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 3079)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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 OR sfGFP
FEATURES             Location/Qualifiers
     source          1..3079
                     /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             847..894
                     /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             895..918
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1057..1075)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1071..1118
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      complement(1336..1924)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1416..1435)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2096..2887,2888..2956))
                     /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     2719..2738
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2957..3061)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gctagtggtg ctagccccgc gaaattaata cgactcacta tagggatgaa cttaagatcg
       61 cgctttatgg ttatgaaaca gaggagacat aacatgaaag cgcgaaacag aacaagaata
      121 agtgcaaatg cttgttctat cgtcttgaaa cagaggagac aagacatgag aacaagcaac
      181 ctggcggcag cgcaaaagat gagcaaaggt gaagaactgt ttaccggcgt tgtgccgatt
      241 ctggtggaac tggatggcga tgtgaacggt cacaaattca gcgtgcgtgg tgaaggtgaa
      301 ggcgatgcca cgattggcaa actgacgctg aaatttatct gcaccaccgg caaactgccg
      361 gtgccgtggc cgacgctggt gaccaccctg acctatggcg ttcagtgttt tagtcgctat
      421 ccggatcaca tgaaacgtca cgatttcttt aaatctgcaa tgccggaagg ctatgtgcag
      481 gaacgtacga ttagctttaa agatgatggc aaatataaaa cgcgcgccgt tgtgaaattt
      541 gaaggcgata ccctggtgaa ccgcattgaa ctgaaaggca cggattttaa agaagatggc
      601 aatatcctgg gccataaact ggaatacaac tttaatagcc ataatgttta tattacggcg
      661 gataaacaga aaaatggcat caaagcgaat tttaccgttc gccataacgt tgaagatggc
      721 agtgtgcagc tggcagatca ttatcagcag aataccccga ttggtgatgg tccggtgctg
      781 ctgccggata atcattatct gagcacgcag accgttctgt ctaaagatcc gaacgaaaaa
      841 ggcacgcggg accacatggt tctgcacgaa tatgtgaatg cggcaggtat tacgtggagc
      901 catccgcagt tcgaaaaata aaggatcccg ggaattctcg agtaaggtta acctgcagga
      961 ggcctttaat taaggtggtg cggccgcgct agcggtcccg ggggatcgat ccggctgcta
     1021 acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac
     1081 cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg
     1141 gaagcttggc actggccgac cggggtcgag cactgactcg ctgcgctcgg tcgttcggct
     1201 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     1261 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     1321 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     1381 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     1441 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     1501 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     1561 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     1621 cgccttatcc ggtaactatc gtcttgagtc caacccgcta agacacgact tatcgccact
     1681 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     1741 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     1801 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     1861 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     1921 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcaca
     1981 gatccgggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     2041 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     2101 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     2161 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     2221 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     2281 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     2341 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     2401 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     2461 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     2521 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     2581 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     2641 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     2701 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     2761 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     2821 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     2881 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     2941 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     3001 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     3061 gcacatttcc ccgaaaagt
//
