LOCUS       sequence-213333-        5078 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-terminal half of pi-EB1 with fast LOV2, no fluorescent tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5078)
  AUTHORS   van Haren J, Charafeddine RA, Ettinger A, Wang H, Hahn KM, Wittmann 
            T
  TITLE     Local control of intracellular microtubule dynamics by EB1 
            photodissociation.
  JOURNAL   Nat Cell Biol. 2018 Jan 29. pii: 10.1038/s41556-017-0028-5. doi: 
            10.1038/s41556-017-0028-5.
  PUBMED    29379139
REFERENCE   2  (bases 1 to 5078)
  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: "Nat Cell 
            Biol. 2018 Jan 29. pii: 10.1038/s41556-017-0028-5. doi: 
            10.1038/s41556-017-0028-5."
COMMENT     Sequence Label: pEB1N-LZ-LOV2(fast)
FEATURES             Location/Qualifiers
     source          1..5078
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        115..418
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        419..622
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     573..593
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1233..1289
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     polyA_signal    1920..2041
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1957..1976)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2011..2030
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2048..2503)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2185..2206)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2397..2416
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2530..2634
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2636..2993
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2636..2827
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2659..2679)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2844..2979
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3028..3822
                     /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             3028..3822
                     /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(3082..3101)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3692..3711
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4010..4029)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4054..4101
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4430..5018
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4919..4938
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta
       61 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      121 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      181 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      241 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      301 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      361 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      421 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      481 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      541 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      601 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc accatggcag
      661 tgaacgtata ctcaacgtca gtgaccagtg ataacctaag tcgacatgac atgctggcct
      721 ggatcaatga gtctctgcag ttgaatctga caaagatcga acagttgtgc tcaggggctg
      781 cgtattgtca gtttatggac atgctgttcc ctggctccat tgccttgaag aaagtgaaat
      841 tccaagctaa gctagaacac gagtacatcc agaacttcaa aatattacaa gcaggtttta
      901 agagaatggg tgttgacaaa ataatcccgg tggacaaatt agtaaaagga aagtttcagg
      961 acaattttga attcgttcag tggttcaaga agttcttcga tgcaaactat gatggaaaag
     1021 actatgaccc tgtggctgct agacaaggtc aagaaactgc agtggctcct tcccttgttg
     1081 ctccagctct gaataaaccg aagaaacctc tcacttctag cagtgcagct ccccagaggc
     1141 ccatctcaac acagagaacc gctgcggctc ctaaggctgg ccctggtgtg gtgcgaaaga
     1201 accctggtat gaaacagctg gaagataaag tggaagagct gttgagcaaa aactatcatc
     1261 tggaaaacga agtggcgcgt ctgaaaaagc tggtgggcga actcgagctc aagcttcgaa
     1321 ttctgcagtc gacggtaccg cggggttctg gaagtggatt ggctgctgca cttgaacgta
     1381 ttgagaagaa ctttgtcatt actgacccaa gattgcctga taatcccgtt atattcgcgt
     1441 ccgatagttt cttgcagttg acagaatata gccgtgaaga aattttggga agaaactgca
     1501 ggtttctaca aggtcctgaa actgatcgcg cgacagtgag aaaaattaga gatgccatag
     1561 ataaccaaac agaggtcact gttcagctga ttaattatac taagagtggt aaaaagttct
     1621 ggaacctctt tcacttgcag cctatgcgag atcagaaggg agatgtccag tactttattg
     1681 gggttcagtt ggatggaact gagcttgtcc gagatgctgc cgagagagag ggagtcatgc
     1741 tgattaagaa aactgcagaa aatatagatg aggcggcaaa agaactttaa ggatccaccg
     1801 gatctagata actgatcata atcagccata ccacatttgt agaggtttta cttgctttaa
     1861 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     1921 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     1981 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     2041 aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     2101 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     2161 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     2221 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     2281 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     2341 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     2401 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     2461 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg
     2521 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     2581 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag
     2641 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     2701 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     2761 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     2821 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     2881 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     2941 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat
     3001 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     3061 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     3121 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     3181 gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc
     3241 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     3301 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     3361 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     3421 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     3481 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     3541 gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     3601 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     3661 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     3721 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     3781 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     3841 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     3901 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     3961 gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga
     4021 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc
     4081 acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg
     4141 ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc
     4201 accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg
     4261 ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat ttttaattta
     4321 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     4381 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     4441 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     4501 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     4561 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     4621 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     4681 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     4741 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     4801 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     4861 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     4921 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     4981 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     5041 tacggttcct ggccttttgc tggccttttg ctcacatg
//
