LOCUS       sequence-194908-        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     19..38
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        19..37
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             121..143
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             163..180
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             184..216
                     /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             256..276
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(1012..1030)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1026..1073
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1170..1625
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1257..1276)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1467..1488
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1651..1755
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1756..1824,1825..2616)
                     /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(1974..1993)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2787..3375
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3276..3295
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3529..3546
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 atctcgatcc cgcgaaatta atacgactca ctatagggag accacaacgg tttccctcta
       61 gaatcaccga gtttattctt gacacctgat gcgatgaatg atataatagg aaagtactgt
      121 tttgtttaac tttaagaagg agatatacat atgcggggtt ctcatcatca tcatcatcat
      181 ggtatggcta gcatgactgg tggacagcaa atgggtcggg atctgtacga cgatgacgat
      241 aacgatccgc tcgaggaaaa cctgtacttc cagggtatga gtgttccaga aaatatgaag
      301 atcaagttgc gcatggaagg ttctgtaaac gggaaccatt tcgtgattga tggaacggga
      361 aaaggaaaac cttacgatgg aacacagact ttagacctga cagtcactaa aggcggacct
      421 ctgcctttcg cttacgatat cttgacaaca gcgttccagt acggcaacag ggcattcacc
      481 gaatacccat cagatatccc agactacttc aagcagactt ttcctaaagg gtataaatgg
      541 gaaagaacca tgacttttga agacaacggc gtgtgtaccg tcaacagcga tataagcatg
      601 gaaggcgact gttttaaata tgaaattcgt tttcatggtg caaattttcc ccccaatggt
      661 ccagttatgc agaagaagac tcagaaatgg gagccatcca ccgagaatat gtatgagtct
      721 gatgggtttc tgaagggtga tgtttgcaga actctgttgc ttaaaggagg cggccattac
      781 gcatgtgact ttaaaagtat ttacaaagcg aaggacagcc acggaatcaa gatggcaaaa
      841 tatcacttta ttgaccaccg cattgagatt ttgaaccatg acgaagatta caacaacgtc
      901 aagctgtatg agaaagcagt tgctcgctat tctatgctgc cgagtaaggc caattgacgt
      961 ttgatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc
     1021 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag
     1081 gaggaactat atccggatct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     1141 acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc
     1201 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     1261 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     1321 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     1381 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     1441 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     1501 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     1561 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     1621 aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     1681 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     1741 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     1801 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     1861 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     1921 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     1981 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     2041 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     2101 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     2161 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     2221 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     2281 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     2341 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     2401 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     2461 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     2521 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     2581 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     2641 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     2701 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     2761 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     2821 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     2881 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     2941 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     3001 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     3061 gactcaagac gatagttacc ggataaggcg cagcggtcgg actgaacggg gggttcgtgc
     3121 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     3181 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     3241 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     3301 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     3361 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     3421 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     3481 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     3541 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     3601 cattaatgca gg
//
