LOCUS       sequence-370625-        3085 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses EGFP from PR promoter in E.coli Dh5alpha cell.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3085)
  AUTHORS   Mukhopadhyay S, Sarkar K, Srivastava R, Pal A, Bagh S
  TITLE     Processing two environmental chemical signals with a synthetic 
            genetic IMPLY gate, a 2-input-2-output integrated logic circuit, and
            a process pipeline to optimize its systems chemistry in Escherichia 
            coli.
  JOURNAL   Biotechnol Bioeng. 2020 May;117(5):1502-1512. doi: 
            10.1002/bit.27286. Epub 2020 Feb 7.
  PUBMED    31981217
REFERENCE   2  (bases 1 to 3085)
  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"; doi: "10.1002/bit.27286";
            journalName: "Biotechnol Bioeng"; date: "2020-05"; volume: "117"; 
            issue: "5"; pages: "1502-1512"
COMMENT     Sequence Label: pNB_EA_3EGFP
FEATURES             Location/Qualifiers
     source          1..3085
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     89..107
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     RBS             192..203
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(210..212,213..215,216..929)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(210..212,213..215,216..926)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(255..276)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(516..535)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     863..884
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      941..1027
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(1300..1888)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1380..1399)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1976..2070)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(2100..2891,2892..2960))
                     /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     2723..2742
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2961..3065)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacctgacgt ctactgggac gaagacgaac agtcgacaac gaagacgcca acatcaagat
       61 ctttattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca
      121 cctcgagtaa caccgtgcgt gttgactatt ttacctctgg cggtgataat ggttgcatgt
      181 acgaattcat taaagaggag aaaggtacca tggtgagcaa gggcgaggag ctgttcaccg
      241 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
      301 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
      361 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
      421 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
      481 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
      541 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
      601 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
      661 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
      721 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
      781 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
      841 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
      901 ctctcggcat ggacgagctg tacaagtaat ctagaggcat caaataaaac gaaaggctca
      961 gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag
     1021 gacaaatccg ccgccctaga cctaggcgat gacggaaaaa gagatcgtct taatgtatag
     1081 attttcgtgg attacgtcgc cagtcaggat cctggcagaa gctattcatt ctgcagttcc
     1141 gtgctccaaa acaaccgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     1201 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     1261 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     1321 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     1381 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     1441 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     1501 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     1561 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     1621 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     1681 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     1741 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     1801 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     1861 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     1921 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgactagtg cttggattct
     1981 caccaataaa aaacgcccgg cggcaaccga gcgttctgaa caaatccaga tggagttctg
     2041 aggtcattac tggatctatc aacaggagtc caagcgagct cgtaaacttg gtctgacagt
     2101 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     2161 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     2221 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     2281 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     2341 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     2401 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     2461 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     2521 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     2581 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     2641 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     2701 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     2761 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     2821 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     2881 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     2941 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     3001 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     3061 ccgcgcacat ttccccgaaa agtgc
//
