LOCUS       sequence-194901-        3599 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the fluorescent protein amFP486/K68M for constitutive 
            expression in E. coli..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3599)
  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 3599)
  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: pThom77.3
FEATURES             Location/Qualifiers
     source          1..3599
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     190..209
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     443..460
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     545..564
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        545..563
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             647..669
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             689..706
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             710..742
                     /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(746..754,755..769)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     complement(1525..1543)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1539..1586
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1683..2138
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1770..1789)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1980..2001
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2164..2268
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2269..2337,2338..3129)
                     /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(2487..2506)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(3300..3599,1..289)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
       61 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      121 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      181 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      241 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      301 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      361 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      421 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      481 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcaggatct cgatcccgcg
      541 aaattaatac gactcactat agggagacca caacggtttc cctctagaat caccgagttt
      601 attcttgaca cctgatgcga tgaatgatat aataggaaag tactgttttg tttaacttta
      661 agaaggagat atacatatgc ggggttctca tcatcatcat catcatggta tggctagcat
      721 gactggtgga cagcaaatgg gtcgggatct gtacgacgat gacgataagg atccggccct
      781 gagcaacaag ttcatcggcg acgacatgaa gatgacctac cacatggacg gctgcgtgaa
      841 cggccactac ttcaccgtga agggcgaggg caacggcaag ccctacgagg gcacccagac
      901 cagcaccttc aaggtgacca tggccaacgg cggccccctg gccttcagct tcgacatcct
      961 gagcaccgtg ttcatgtacg gcaaccggtg cttcaccgcc taccccacca gcatgcccga
     1021 ctacttcaag caggccttcc ccgacggcat gagctacgag cggaccttca cctacgagga
     1081 cggcggcgtg gccaccgcca gctgggagat cagcctgaag ggcaactgct tcgagcacaa
     1141 gagcaccttc cacggcgtga acttccccgc cgacggcccc gtgatggcca agaagaccac
     1201 cggctgggac cccagcttcg agaagatgac cgtgtgcgac ggcatcctga agggcgacgt
     1261 gaccgccttc ctgatgctgc agggcggcgg caactaccgg tgccagttcc acaccagcta
     1321 caagaccaag aagcccgtga ccatgccccc caaccacgtg gtggagcacc ggatcgcccg
     1381 gaccgacctg gacaagggcg gcaacagcgt gcagctgacc gagcacgccg tggcccacat
     1441 caccagcgtg gtgcccttct aagaattcga agcttgatcc ggctgctaac aaagcccgaa
     1501 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct
     1561 ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga tctggcgtaa
     1621 tagcgaagag gccagcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     1681 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     1741 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     1801 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     1861 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     1921 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     1981 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     2041 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     2101 taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat
     2161 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     2221 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     2281 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     2341 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     2401 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     2461 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     2521 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     2581 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     2641 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     2701 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     2761 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     2821 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     2881 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     2941 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     3001 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     3061 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     3121 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     3181 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     3241 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     3301 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     3361 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     3421 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     3481 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     3541 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataa
//
