LOCUS       sequence-199156-        3612 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the fluorescent protein EG-4 for constitutive expression in 
            E. coli..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3612)
  AUTHORS   Molina RS, Tran TM, Campbell RE, Lambert GG, Salih A, Shaner NC, 
            Hughes TE, Drobizhev M
  TITLE     Blue-Shifted Green Fluorescent Protein Homologues Are Brighter than 
            Enhanced Green Fluorescent Protein under Two-Photon Excitation.
  JOURNAL   J Phys Chem Lett. 2017 Jun 15;8(12):2548-2554. doi: 
            10.1021/acs.jpclett.7b00960. Epub 2017 May 25.
  PUBMED    28530831
REFERENCE   2  (bases 1 to 3612)
  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: 
            "10.1021/acs.jpclett.7b00960"; journalName: "J Phys Chem Lett"; 
            date: "2017-06-15- 15"; volume: "8"; issue: "12"; pages: "2548-2554"
COMMENT     Sequence Label: pRSM15
FEATURES             Location/Qualifiers
     source          1..3612
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..39
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        20..38
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             122..144
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             164..181
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             185..217
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             257..277
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(1013..1031)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1027..1074
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1171..1626
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1258..1277)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1468..1489
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1652..1756
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1757..1825,1826..2617)
                     /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     complement(1975..1994)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2788..3376
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3277..3296
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3530..3547
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaatcaccg agtttattct tgacacctga tgcgatgaat gatataatag gaaagtactg
      121 ttttgtttaa ctttaagaag gagatataca tatgcggggt tctcatcatc atcatcatca
      181 tggtatggct agcatgactg gtggacagca aatgggtcgg gatctgtacg acgatgacga
      241 taacgatccg ctcgaggaaa acctgtactt ccagggtatg agtgttccag aaaatatgaa
      301 gatcaagttg cgcatggaag gttctgtaaa cgggaaccat ttcgtgattg atggaacggg
      361 aaaaggaaaa ccttacgatg gaacacagac tttagacctg acagtcacta aaggcggacc
      421 tctgcctttc gcttacgata tcttgacaac agcgttccag tacggcaaca gggcattcac
      481 cgaataccca tcagatatcc cagactactt caagcagact tttcctaaag ggtataaatg
      541 ggaaagaacc atgacttttg aagacaacgg cgtgtgtacc gtcaacagcg atataagcat
      601 ggaaggcgac tgttttaaat atgaaattcg ttttcatggt gcaaattttc cccccaatgg
      661 tccagttatg cagaagaaga ctcagaaatg ggagccatcc accgagaata tgtatgagtc
      721 tgatgggttt ctgaagggtg atgtttgcag aactctgttg cttaaaggag gcggccatta
      781 cgcatgtgac tttaaaagta tttacaaagc gaaggacagc cacggaatca agatggcaaa
      841 atatcacttt attgaccacc gcattgagat tttgaaccat gacgaagatt acaacaacgt
      901 caagctgtat gagaaagcag ttgctcgcta ttctatgctg ccgagtaagg ccaattgacg
      961 tttgatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag
     1021 caataactag cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa
     1081 ggaggaacta tatccggatc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     1141 aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg
     1201 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     1261 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     1321 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     1381 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     1441 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     1501 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     1561 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     1621 caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     1681 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     1741 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     1801 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     1861 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     1921 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     1981 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     2041 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     2101 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     2161 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     2221 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     2281 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     2341 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     2401 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     2461 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     2521 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     2581 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     2641 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     2701 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     2761 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     2821 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     2881 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     2941 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3001 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3061 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3121 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     3181 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     3241 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     3301 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     3361 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     3421 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     3481 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     3541 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     3601 tcattaatgc ag
//
