LOCUS       sequence-204391-        3013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses sfGFP (a GFP) in Ecoli off a T7 promoter when pNP1 AND 
            input A and pNP1 AND input B are co-expressed.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3013)
  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 3013)
  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 AND gate sfGFP
FEATURES             Location/Qualifiers
     source          1..3013
                     /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             781..828
                     /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             829..852
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(991..1009)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1005..1052
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      complement(1270..1858)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1350..1369)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2030..2821,2822..2890))
                     /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     2653..2672
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2891..2995)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gctagtggtg ctagccccgc gaaattaata cgactcacta tagggcttat gagtgtaata
       61 cgttctatgt cagattcaag aacagaggag attgaaatgg acatagaacg aaacctggcg
      121 gcagcgcaaa agatgagcaa aggtgaagaa ctgtttaccg gcgttgtgcc gattctggtg
      181 gaactggatg gcgatgtgaa cggtcacaaa ttcagcgtgc gtggtgaagg tgaaggcgat
      241 gccacgattg gcaaactgac gctgaaattt atctgcacca ccggcaaact gccggtgccg
      301 tggccgacgc tggtgaccac cctgacctat ggcgttcagt gttttagtcg ctatccggat
      361 cacatgaaac gtcacgattt ctttaaatct gcaatgccgg aaggctatgt gcaggaacgt
      421 acgattagct ttaaagatga tggcaaatat aaaacgcgcg ccgttgtgaa atttgaaggc
      481 gataccctgg tgaaccgcat tgaactgaaa ggcacggatt ttaaagaaga tggcaatatc
      541 ctgggccata aactggaata caactttaat agccataatg tttatattac ggcggataaa
      601 cagaaaaatg gcatcaaagc gaattttacc gttcgccata acgttgaaga tggcagtgtg
      661 cagctggcag atcattatca gcagaatacc ccgattggtg atggtccggt gctgctgccg
      721 gataatcatt atctgagcac gcagaccgtt ctgtctaaag atccgaacga aaaaggcacg
      781 cgggaccaca tggttctgca cgaatatgtg aatgcggcag gtattacgtg gagccatccg
      841 cagttcgaaa aataaaggat cccgggaatt ctcgagtaag gttaacctgc aggaggcctt
      901 taattaaggt ggtgcggccg cgctagcggt cccgggggat cgatccggct gctaacaaag
      961 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
     1021 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccggaagct
     1081 tggcactggc cgaccggggt cgagcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     1141 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     1201 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     1261 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     1321 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     1381 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     1441 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     1501 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     1561 atccggtaac tatcgtcttg agtccaaccc gctaagacac gacttatcgc cactggcagc
     1621 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     1681 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     1741 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     1801 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     1861 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacagatccg
     1921 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     1981 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     2041 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     2101 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     2161 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     2221 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     2281 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     2341 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     2401 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     2461 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     2521 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     2581 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     2641 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     2701 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     2761 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     2821 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     2881 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     2941 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     3001 ttccccgaaa agt
//
