LOCUS       sequence-194943-        3533 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the fluorescent protein amFP486 for constitutive expression 
            in E. coli..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3533)
  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 3533)
  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: pThom77.2
FEATURES             Location/Qualifiers
     source          1..3533
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(60..78)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      74..121
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      218..673
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(305..324)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     515..536
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        699..803
                     /gene="bla"
                     /label=AmpR promoter
     CDS             804..1598
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             804..1598
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(858..877)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1468..1487
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      1769..2357
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2258..2277
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2511..2528
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     2613..2632
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2613..2631
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2715..2737
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2757..2774
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2778..2810
                     /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(2814..2822,2823..2837)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
ORIGIN
        1 ttcgaagctt gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac
       61 cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
      121 gctgaaagga ggaactatat ccggatctgg cgtaatagcg aagaggccag caccgatcgc
      181 ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta
      241 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
      301 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
      361 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
      421 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
      481 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
      541 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
      601 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
      661 acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
      721 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
      781 aataatattg aaaaaggaag agtatgattg aacaagatgg attgcacgca ggttctccgg
      841 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
      901 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
      961 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
     1021 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     1081 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     1141 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     1201 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     1261 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     1321 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     1381 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     1441 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     1501 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     1561 gcatcgcctt ctatcgcctt cttgacgagt tcttctgact gtcagaccaa gtttactcat
     1621 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     1681 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     1741 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     1801 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     1861 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     1921 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     1981 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     2041 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     2101 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     2161 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     2221 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     2281 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     2341 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     2401 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     2461 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     2521 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     2581 ttcattaatg caggatctcg atcccgcgaa attaatacga ctcactatag ggagaccaca
     2641 acggtttccc tctagaatca ccgagtttat tcttgacacc tgatgcgatg aatgatataa
     2701 taggaaagta ctgttttgtt taactttaag aaggagatat acatatgcgg ggttctcatc
     2761 atcatcatca tcatggtatg gctagcatga ctggtggaca gcaaatgggt cgggatctgt
     2821 acgacgatga cgataaggat ccggccctga gcaacaagtt catcggcgac gacatgaaga
     2881 tgacctacca catggacggc tgcgtgaacg gccactactt caccgtgaag ggcgagggca
     2941 acggcaagcc ctacgagggc acccagacca gcaccttcaa ggtgaccatg gccaacggcg
     3001 gccccctggc cttcagcttc gacatcctga gcaccgtgtt caagtacggc aaccggtgct
     3061 tcaccgccta ccccaccagc atgcccgact acttcaagca ggccttcccc gacggcatga
     3121 gctacgagcg gaccttcacc tacgaggacg gcggcgtggc caccgccagc tgggagatca
     3181 gcctgaaggg caactgcttc gagcacaaga gcaccttcca cggcgtgaac ttccccgccg
     3241 acggccccgt gatggccaag aagaccaccg gctgggaccc cagcttcgag aagatgaccg
     3301 tgtgcgacgg catcctgaag ggcgacgtga ccgccttcct gatgctgcag ggcggcggca
     3361 actaccggtg ccagttccac accagctaca agaccaagaa gcccgtgacc atgcccccca
     3421 accacgtggt ggagcaccgg atcgcccgga ccgacctgga caagggcggc aacagcgtgc
     3481 agctgaccga gcacgccgtg gcccacatca ccagcgtggt gcccttctaa gaa
//
