LOCUS       sequence-308719-        5119 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  1-100 amino acids of Arabidopsis PIF6 (Gene ID: 825382), tagged with
            EGFP fluorescent protein. Mammalian codon optimised..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5119)
  AUTHORS   Buckley CE, Moore RE, Reade A, Goldberg AR, Weiner OD, Clarke JD
  TITLE     Reversible Optogenetic Control of Subcellular Protein Localization 
            in a Live Vertebrate Embryo.
  JOURNAL   Dev Cell. 2016 Jan 11;36(1):117-26. doi: 
            10.1016/j.devcel.2015.12.011.
  PUBMED    26766447
REFERENCE   2  (bases 1 to 5119)
  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: 
            "10.1016/j.devcel.2015.12"; journalName: "Dev Cell"; date: 
            "2016-01-11- 11"; volume: "36"; issue: "1"; pages: "117-26"
COMMENT     Sequence Label: pCS2+-PIF6-EGFP
FEATURES             Location/Qualifiers
     source          1..5119
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             join(448..450,451..1164)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(448..450,451..1161)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(490..511)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(751..770)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1098..1119
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1171..1305
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1195..1214)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1249..1268
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1418..1438)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1418..1436)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1457..1473)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1457..1473)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1470..1492)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1481..1497
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1505..1511,1512..1529,1530..1535))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1550..1571
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1688..1705)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1859..2447)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1939..1958)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2618..3409,3410..3478))
                     /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     3241..3260
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3479..3583)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3609..4064)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3746..3767)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3958..3977
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4135..5119
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcga ttcgaattcg ccaccatgat gttccttcct
      121 actgattatt gttgcagact gagcgaccag gaatacatgg aactcgtctt cgagaacgga
      181 cagatactcg caaaaggcca gaggtcaaat gttagtctcc ataatcagcg gacgaaaagc
      241 atcatggatc tgtatgaggc cgaatacaac gaagatttta tgaaaagtat tatccatgga
      301 gggggtggcg ctattaccaa cctgggagat acccaagtgg tcccacagtc ccacgtagca
      361 gccgctcacg agaccaatat gctggagtcc aacaaacacg tagacacgcg tgatatcgat
      421 tccgcaggct ctgctggctc agcaggcgtg agcaagggcg aggagctgtt caccggggtg
      481 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
      541 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
      601 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
      661 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
      721 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
      781 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
      841 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
      901 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
      961 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     1021 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     1081 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     1141 ggcatggacg agctgtacaa gtaatacgta gatccagaca tgataagata cattgatgag
     1201 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     1261 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     1321 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaatt cgcggccgcg
     1381 gcgccaatgc attgggcccg gtacccagct tttgttccct ttagtgaggg ttaattgcgc
     1441 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     1501 cacacaacat acgagccggg agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     1561 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     1621 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     1681 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     1741 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     1801 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     1861 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     1921 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     1981 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2041 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     2101 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2161 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2221 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2281 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2341 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     2401 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2461 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     2521 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     2581 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     2641 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     2701 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     2761 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     2821 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     2881 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     2941 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     3001 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     3061 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     3121 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     3181 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     3241 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     3301 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     3361 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     3421 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     3481 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     3541 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     3601 tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     3661 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     3721 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     3781 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     3841 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     3901 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     3961 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     4021 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccattcag
     4081 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagtcgac
     4141 cgccaattca atatggcgta tatggactca tgccaattca atatggtgga tctggacctg
     4201 tgccaattca atatggcgta tatggactcg tgccaattca atatggtgga tctggacccc
     4261 agccaattca atatggcgga cttggcacca tgccaattca atatggcgga cctggcactg
     4321 tgccaactgg ggaggggtct acttggcacg gtgccaagtt tgaggagggg tcttggccct
     4381 gtgccaagtc cgccatattg aattggcatg gtgccaataa tggcggccat attggctata
     4441 tgccaggatc aatatatagg caatatccaa tatggcccta tgccaatatg gctattggcc
     4501 aggttcaata ctatgtattg gccctatgcc atatagtatt ccatatatgg gttttcctat
     4561 tgacgtagat agcccctccc aatgggcggt cccatatacc atatatgggg cttcctaata
     4621 ccgcccatag ccactccccc attgacgtca atggtctcta tatatggtct ttcctattga
     4681 cgtcatatgg gcggtcctat tgacgtatat ggcgcctccc ccattgacgt caattacggt
     4741 aaatggcccg cctggctcaa tgcccattga cgtcaatagg accacccacc attgacgtca
     4801 atgggatggc tcattgccca ttcatatccg ttctcacgcc ccctattgac gtcaatgacg
     4861 gtaaatggcc cacttggcag tacatcaata tctattaata gtaacttggc aagtacatta
     4921 ctattggaag tacgccaggg tacattggca gtactcccat tgacgtcaat ggcggtaaat
     4981 ggcccgcgat ggctgccaag tacatcccca ttgacgtcaa tggggagggg caatgacgca
     5041 aatgggcgtt ccattgacgt aaatgggcgg taggcgtgcc taatgggagg tctatataag
     5101 caatgctcgt ttagggaac
//
