LOCUS       sequence-190275-        7899 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 7899)
  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 7899)
  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..7899
                     /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"
     primer_bind     complement(4470..4489)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4471..4489)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4576..4593)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4747..5335)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4827..4846)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5506..6297,6298..6366))
                     /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     6129..6148
                     /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 atwaymawat mattwskgka tkcatcagcg ttaagcatgt
     4261 aataattaac atgtaatgca tgacgttatt tatgagatgg gtttttatga ttagagtccc
     4321 gcaattatac atttaatacg cgatagaaaa caaaatatag cgcgcaaact aggataaatt
     4381 atcgcgcgcg gtgtcatcta tgttactaga tccgatgata agctgtcaaa catgagaatt
     4441 cttatcgatg atatcagatc tgccggtctc cctatagtga gtcgtattaa tttcgataag
     4501 ccaggttaac ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     4561 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4621 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4681 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4741 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4801 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4861 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4921 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4981 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     5041 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5101 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5161 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     5221 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5281 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5341 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5401 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     5461 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     5521 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     5581 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     5641 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     5701 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     5761 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     5821 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     5881 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     5941 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     6001 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     6061 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     6121 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     6181 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     6241 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     6301 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     6361 actcatactc ttcctttttc aatatcgagc acgttgatgg gatcgatcct agaatatcaa
     6421 tcgagccacc gtcaatttcg agcgctaagt ggagttgtcc gagtcccgaa tcacaagaat
     6481 taggctcgtg ctctgtgaga tctctcgcga aacccagatg aaggaaaaaa atcggaagat
     6541 cgcggaagaa gtggggttcg catggtctaa cattgtcgca ttcctcacag tttcggctgg
     6601 aaacggcagg gacaatcacg agaaatgcgc gacgagtatt cagttgtcca aaattatgtc
     6661 agcttgaaag ttggaaacgg cggaagaaaa ttcgtcagga gggcgttgcg gctagggtga
     6721 agaacggaga caatttcggc tttgaaattg gctcgactct gattggatca tgacgcactg
     6781 ctggtccaca accgtgtcga aggtgccgtc tttactacag gtccgctgga agcaaaatgg
     6841 aaaaagccgc tggagcccga ctacagagcc gccgtgtttt ggtaatcagt cggcaaatag
     6901 gtcagcacag cgcagcgtga caggttcttg caatttacag cacagctcgt ccgtctacga
     6961 ctttgcacac cacaaagtgt gcggggagca aaggagccga tcttggtcgc gcgcaaagcc
     7021 aaggagtctt gaacctgaga gtgtgcgtgt cttgtgacgc ttgcccttct gtactttgct
     7081 gtgacactac caccacatct gtcttggctt tttgttcata catccacacc gaccatgtcg
     7141 ctattcaacg tcagcaacgg tcttcgtacc gctctccgac cttctgttgc cagctcttcg
     7201 cgcgttgctg ccttttccac aaccgccgct gcccgtctcg ccacacccac ctctgacaac
     7261 gttggcagtt cgggcaagcc tcagcacttg aagcagttca agatctaccg atggaaccct
     7321 gacaagccct cggagaagcc tcgtctgcag tcgtacacac tggacctcaa ccagaccggt
     7381 ccaatggttc tcgacgcgct catcaaaatc aagaacgaaa ttgaccctac gctcaccttc
     7441 cgtcgctcgt gccgtgaggg tatctgcggt tcgtgcgcta tgaatattga cggtgtcaac
     7501 accctcgcct gcctctgccg gatcgacaag cagaatgaca ccaagatcta ccccttgccg
     7561 cacatgtaca ttgtcaagga cctcgtgcca gacttgaccc agttctacaa gcagtaccga
     7621 tccatcgagc ctttcctcaa gtccaacaac accccttctg agggtgaaca tcttcagtcg
     7681 cccgaggagc gtcgtcgact cgacggtctg tacgagtgca ttctgtgcgc gtgctgctcc
     7741 acatcctgcc cctcttactg gtggaatcag gacgagtacc ttggccccgc cgtgctcatg
     7801 caggcgtacc gatggatggc cgactcgcgt gacgactttg gtgaggagcg aagacagaag
     7861 ctcgagaaca ccttttcgct ctaccgatgc cttaccatc
//
