LOCUS       sequence-204602-        3602 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the fluorescent protein eqFP486 for constitutive expression 
            in E. coli..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3602)
  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 3602)
  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: pRSM17
FEATURES             Location/Qualifiers
     source          1..3602
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      8..596
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     497..516
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     750..767
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     852..871
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        852..870
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             954..976
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             996..1013
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1017..1049
                     /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             join(1053..1061,1062..1076)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     complement(1835..1853)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1849..1896
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1993..2448
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2080..2099)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2290..2311
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2474..2578
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2579..2647,2648..3439)
                     /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(2797..2816)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
       61 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
      121 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
      181 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
      241 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
      301 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
      361 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
      421 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
      481 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
      541 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
      601 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
      661 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
      721 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
      781 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc aggatctcga
      841 tcccgcgaaa ttaatacgac tcactatagg gagaccacaa cggtttccct ctagaatcac
      901 cgagtttatt cttgacacct gatgcgatga atgatataat aggaaagtac tgttttgttt
      961 aactttaaga aggagatata catatgcggg gttctcatca tcatcatcat catggtatgg
     1021 ctagcatgac tggtggacag caaatgggtc gggatctgta cgacgatgac gataaggatc
     1081 cggatcccct agtcaacggc aacctgaaga tcaaaatgtt tatgaatggc agcgtgaaca
     1141 agcacaaatt caaatgtgag gcagagggag aaggaaatat ccatagcggc acccatatta
     1201 tgaagattac cgtgactgag ggaggcccgc tgccttttgc cttcgacata ctgtcccgga
     1261 ccttctgcta tggtcacaaa atgttcctgg acaacagcgg gaatatccca gacttcttca
     1321 aacagagcct taaggacggg tactcttggg agagagaaac gacctacgag gacggcggag
     1381 tactgaatag taaacagaag cacactctga agggcgactg catcacaact tattctgaag
     1441 ttaccggaac atttccccct aacggcccag tgatgaaaaa aaagaccctc ggctgggagc
     1501 cctcctgcga aatggtcatc ccccgggatg ggggtttgat catgtacgat aatattgccc
     1561 tgaagcttga tgacgatcac ggcgggcacc tgaccgggaa ttttaaggcc acatacaagt
     1621 caaataaccc caacatcaac ctccctaaac accatttcgt ggagaaccgc gtggagatgt
     1681 tagaaggtcg ggacggcggc aattatgtga agctgcatga aaggaacaag gcccggtaca
     1741 gcgccttgga gagcgagctc gctaagaagt aggaattcga agcttgatcc ggctgctaac
     1801 aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc
     1861 cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga
     1921 tctggcgtaa tagcgaagag gccagcaccg atcgcccttc ccaacagttg cgcagcctga
     1981 atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     2041 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     2101 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     2161 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     2221 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     2281 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     2341 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     2401 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt
     2461 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     2521 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     2581 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     2641 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     2701 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     2761 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     2821 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     2881 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     2941 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     3001 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     3061 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     3121 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     3181 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     3241 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     3301 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     3361 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     3421 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     3481 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     3541 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     3601 ag
//
