LOCUS       sequence-209378-        5756 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-terminal half of pi-EB1 with wild-type LOV2, mCherry-tagged to 
            track microtubule ends.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5756)
  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 5756)
  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-mCherry-LOV2(wt)
FEATURES             Location/Qualifiers
     source          1..5756
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1179..1235
                     /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"
     CDS             1254..1961
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1399..1417)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1667..1686
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2544..2665
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2581..2600)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2635..2654
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2672..3127)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2809..2830)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3021..3040
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3154..3258
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3260..3617
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3260..3451
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3283..3303)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3468..3603
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3652..4446
                     /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             3652..4446
                     /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(3706..3725)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4316..4335
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4634..4653)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4678..4725
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5054..5642
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5543..5562
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcacca
      601 tggcagtgaa cgtatactca acgtcagtga ccagtgataa cctaagtcga catgacatgc
      661 tggcctggat caatgagtct ctgcagttga atctgacaaa gatcgaacag ttgtgctcag
      721 gggctgcgta ttgtcagttt atggacatgc tgttccctgg ctccattgcc ttgaagaaag
      781 tgaaattcca agctaagcta gaacacgagt acatccagaa cttcaaaata ttacaagcag
      841 gttttaagag aatgggtgtt gacaaaataa tcccggtgga caaattagta aaaggaaagt
      901 ttcaggacaa ttttgaattc gttcagtggt tcaagaagtt cttcgatgca aactatgatg
      961 gaaaagacta tgaccctgtg gctgctagac aaggtcaaga aactgcagtg gctccttccc
     1021 ttgttgctcc agctctgaat aaaccgaaga aacctctcac ttctagcagt gcagctcccc
     1081 agaggcccat ctcaacacag agaaccgctg cggctcctaa ggctggccct ggtgtggtgc
     1141 gaaagaaccc tggtatgaaa cagctggaag ataaagtgga agagctgttg agcaaaaact
     1201 atcatctgga aaacgaagtg gcgcgtctga aaaagctggt gggcgaactc gagatggtga
     1261 gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca
     1321 tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
     1381 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct
     1441 gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg
     1501 acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga
     1561 acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt
     1621 tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga
     1681 agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga
     1741 agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca
     1801 agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca
     1861 agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg
     1921 agggccgcca ctccaccggc ggcatggacg agctgtacaa gccgcggggt tctggaagtg
     1981 gattggctgc tgcacttgaa cgtattgaga agaactttgt cattactgac ccaagattgc
     2041 cagataatcc cattatattc gcgtccgata gtttcttgca gttgacagaa tatagccgtg
     2101 aagaaatttt gggaagaaac tgcaggtttc tacaaggtcc tgaaactgat cgcgcgacag
     2161 tgagaaaaat tagagatgcc atagataacc aaacagaggt cactgttcag ctgattaatt
     2221 atacaaagag tggtaaaaag ttctggaacc tctttcactt gcagcctatg cgagatcaga
     2281 agggagatgt ccagtacttt attggggttc agttggatgg aactgagcat gtccgagatg
     2341 ctgccgagag agagggagtc atgctgatta agaaaactgc agaaaatatt gatgaggcgg
     2401 caaaagaact ttaaggatcc accggatcta gataactgat cataatcagc cataccacat
     2461 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     2521 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     2581 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     2641 tgtccaaact catcaatgta tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa
     2701 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     2761 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     2821 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     2881 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     2941 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     3001 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     3061 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     3121 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     3181 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     3241 tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     3301 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3361 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3421 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3481 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     3541 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3601 gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa
     3661 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     3721 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     3781 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag
     3841 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     3901 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     3961 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     4021 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     4081 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     4141 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
     4201 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     4261 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     4321 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     4381 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     4441 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     4501 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     4561 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
     4621 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     4681 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
     4741 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     4801 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     4861 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     4921 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     4981 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     5041 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     5101 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     5161 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     5221 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     5281 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     5341 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     5401 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     5461 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     5521 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     5581 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     5641 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     5701 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcat
//
