LOCUS       sequence-174820-        7891 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses lambda N protein fused to triple Gfp coupled to an NLS 
            sequence under the control of the constitutively active Potef 
            promotor..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7891)
  AUTHORS   Pohlmann T, Baumann S, Haag C, Albrecht M, Feldbrugge M
  TITLE     A FYVE zinc finger domain protein specifically links mRNA transport 
            to endosome trafficking.
  JOURNAL   Elife. 2015 May 18;4. doi: 10.7554/eLife.06041.
  PUBMED    25985087
REFERENCE   2  (bases 1 to 7891)
  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"; journalName: "Elife."; 
            date: "2015-05-18"; volume: "4. doi"; pages: " 10.7554/eLife.06041"
COMMENT     Sequence Label: pPotef-LN22*-3xGNLS-cbx
FEATURES             Location/Qualifiers
     source          1..7891
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     570..588
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(626..648)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     748..767
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        848..866
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     848..865
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    join(944..962,963..985,986..1004,1005..1027,1028..1046,
                     1047..1069,1070..1088,1089..1111,1112..1130,1131..1153,
                     1154..1172,1173..1195,1196..1214)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     protein_bind    join(1251..1269,1270..1292,1293..1311,1312..1334,1335..1353,
                     1354..1376,1377..1395,1396..1418,1419..1437,1438..1460,
                     1461..1479,1480..1502,1503..1521)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     regulatory      1899..1908
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1905..1907,1908..1910,1911..2621)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1905..1907,1908..1910,1911..2621)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2622..2624,2625..2627,2628..3338)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2622..2624,2625..2627,2628..3338)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3339..3341,3342..3344,3345..4055)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3339..3341,3342..3344,3345..4055)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             4107..4127
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      4158..4410
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(4466..4485)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4467..4485)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4572..4589)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4743..5331)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4823..4842)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5800..6176)
                     /label=ISS
                     /note="immunostimulatory sequence from the AmpR gene; 
                     contains unmethylated CpG dinucleotides in the context of 
                     5'-AACGTT-3' (Sato et al., 1996)"
     primer_bind     6125..6144
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 atgaactgct ccaggacctg ccccaagaac ctcaaccctg gtaaggcaat tgcacagatc
       61 aagaaggaca tggccgtcgg cgcacccaag gcttccgagc gccctatcat ggcttcgtcg
      121 taatcttgat atatcatatc gttctttcct cagcacttct tttgtcaatt tcaaaagtat
      181 ctaattgcat tcaactccgc ttgtggtttg ttgttcagtg agagtggaaa cgctacgggc
      241 aagatgaggg cagtgttctg gcgacggaaa agtgtgcaag tgtctggcct gcgtcctcgc
      301 tggttccagc agccgatgca ggacgtgtac ctagcgattt cttcgacagc ctattgtggc
      361 agccgcgatt cgccacaatc gtacgaaagc gagacgagtt gagcgaagat ccccattatt
      421 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
      481 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
      541 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
      601 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
      661 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
      721 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
      781 atatggacat attgtcgtta gaacgcggct acaattaata cataacctta tgtatcatac
      841 acatacgatt taggtgacac tatagaactc gagcagctga agcttgcatg cctgcaggtc
      901 gaaattcgag ctcggtaccc gtaccgagct cgactttcac ttttctctat cactgatagg
      961 gagtggtaaa ctcgactttc acttttctct atcactgata gggagtggta aactcgactt
     1021 tcacttttct ctatcactga tagggagtgg taaactcgac tttcactttt ctctatcact
     1081 gatagggagt ggtaaactcg actttcactt ttctctatca ctgataggga gtggtaaact
     1141 cgactttcac ttttctctat cactgatagg gagtggtaaa ctcgactttc acttttctct
     1201 atcactgata gggagtggta aactcgagta ccgagctcga ctttcacttt tctctatcac
     1261 tgatagggag tggtaaactc gactttcact tttctctatc actgataggg agtggtaaac
     1321 tcgactttca cttttctcta tcactgatag ggagtggtaa actcgacttt cacttttctc
     1381 tatcactgat agggagtggt aaactcgact ttcacttttc tctatcactg atagggagtg
     1441 gtaaactcga ctttcacttt tctctatcac tgatagggag tggtaaactc gactttcact
     1501 tttctctatc actgataggg agtggtaaac tcgaggggat caattcgacc aatgaggcgc
     1561 gagacgaggg gacgctggaa gttgaggcgc aagaaaattt ttctctggtt ctgcgcggca
     1621 gagacgacca gattcgcccg ctttcttctg cgttgggtgc ctcttttggg tgccagactt
     1681 tgtgtgtgcg ccagcgagac gttccaataa agggcgctgt ctcggcacta tctttctttc
     1741 tttcctcata catcgtatca taccatacac agacaacatc atccacggga tcccccgggc
     1801 tgcaggaatt cgatcccatg ggcaacgctc gcacgcgtcg tcgcgagcgt cgtgccgaga
     1861 agcaggcgca gtggaaggcc gccaacggtg gatccgcggc cgccatggtg agcaagggcg
     1921 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     1981 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     2041 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     2101 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     2161 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     2221 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     2281 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     2341 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     2401 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     2461 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     2521 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     2581 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gatggtgagc aagggcgagg
     2641 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     2701 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     2761 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     2821 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     2881 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     2941 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     3001 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     3061 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     3121 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     3181 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     3241 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     3301 ccgccgggat cactctcggc atggacgagc tgtacaagat ggtgagcaag ggcgaggagc
     3361 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     3421 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     3481 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     3541 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     3601 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     3661 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     3721 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     3781 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     3841 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     3901 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
     3961 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     4021 ccgggatcac tctcggcatg gacgagctgt acaagagaaa taattttgtt tcactttcag
     4081 ctggcgatat acatatgaaa gcgcctccaa aaaagaagag aaaggtcgaa aaagcgtaaa
     4141 gcggccgccc ggctgcagat cgttcaaaca tttggcaata aagtttctta agattgaatc
     4201 ctgttgccgg tcttgcgatg attatcatat aatttctgtt gaattacgtt aagcatgtaa
     4261 taattaacat gtaatgcatg acgttattta tgagatgggt ttttatgatt agagtcccgc
     4321 aattatacat ttaatacgcg atagaaaaca aaatatagcg cgcaaactag gataaattat
     4381 cgcgcgcggt gtcatctatg ttactagatc cgatgataag ctgtcaaaca tgagaattca
     4441 tcgatgatat cagatctgcc ggtctcccta tagtgagtcg tattaatttc gataagccag
     4501 gttaacctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     4561 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     4621 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     4681 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4741 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4801 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4861 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4921 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4981 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     5041 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     5101 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     5161 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     5221 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     5281 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     5341 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     5401 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     5461 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     5521 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     5581 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     5641 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     5701 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     5761 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     5821 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     5881 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     5941 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     6001 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     6061 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     6121 cgggataata ccgcgccaca tagcagactt taaaagtgct catcattgga aacgttcttc
     6181 ggggcgaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6241 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaacag
     6301 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6361 tcttcctttt tcaatatcga gcacgttgat gggatcgatc ctagaatatc aatcgagcca
     6421 ccgtcaattt cgagcgctaa gtggagttgt ccgagtcccg aatcacaaga attaggctcg
     6481 tgctctgtga gatctctcgc gaaacccaga tgaaggaaaa aaatcggaag atcgcggaag
     6541 aagtggggtt cgcatggtct aacattgtcg cattcctcac agtttcggct ggaaacggca
     6601 gggacaatca cgagaaatgc gcgacgagta ttcagttgtc caaaattatg tcagcttgaa
     6661 agttggaaac ggcggaagaa aattcgtcag gagggcgttg cggctagggt gaagaacgga
     6721 gacaatttcg gctttgaaat tggctcgact ctgattggat catgacgcac tgctggtcca
     6781 caaccgtgtc gaaggtgccg tctttactac aggtccgctg gaagcaaaat ggaaaaagcc
     6841 gctggagccc gactacagag ccgccgtgtt ttggtaatca gtcggcaaat aggtcagcac
     6901 agcgcagcgt gacaggttct tgcaatttac agcacagctc gtccgtctac gactttgcac
     6961 accacaaagt gtgcggggag caaaggagcc gatcttggtc gcgcgcaaag ccaaggagtc
     7021 ttgaacctga gagtgtgcgt gtcttgtgac gcttgccctt ctgtactttg ctgtgacact
     7081 accaccacat ctgtcttggc tttttgttca tacatccaca ccgaccatgt cgctattcaa
     7141 cgtcagcaac ggtcttcgta ccgctctccg accttctgtt gccagctctt cgcgcgttgc
     7201 tgccttttcc acaaccgccg ctgcccgtct cgccacaccc acctctgaca acgttggcag
     7261 ttcgggcaag cctcagcact tgaagcagtt caagatctac cgatggaacc ctgacaagcc
     7321 ctcggagaag cctcgtctgc agtcgtacac actggacctc aaccagaccg gtccaatggt
     7381 tctcgacgcg ctcatcaaaa tcaagaacga aattgaccct acgctcacct tccgtcgctc
     7441 gtgccgtgag ggtatctgcg gttcgtgcgc tatgaatatt gacggtgtca acaccctcgc
     7501 ctgcctctgc cggatcgaca agcagaatga caccaagatc taccccttgc cgcacatgta
     7561 cattgtcaag gacctcgtgc cagacttgac ccagttctac aagcagtacc gatccatcga
     7621 gcctttcctc aagtccaaca acaccccttc tgagggtgaa catcttcagt cgcccgagga
     7681 gcgtcgtcga ctcgacggtc tgtacgagtg cattctgtgc gcgtgctgct ccacatcctg
     7741 cccctcttac tggtggaatc aggacgagta ccttggcccc gccgtgctca tgcaggcgta
     7801 ccgatggatg gccgactcgc gtgacgactt tggtgaggag cgaagacaga agctcgagaa
     7861 caccttttcg ctctaccgat gccttaccat c
//
