LOCUS       sequence-377726-        6216 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses EB1-tdTomato fusion in mammalian cells for labeling the 
            growing tips of MTs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6216)
  TITLE     Dent Lab Unpublished Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6216)
  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: 
            "Unpublished"
COMMENT     Sequence Label: EB1-tdTomato
FEATURES             Location/Qualifiers
     source          1..6216
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1386..1395
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1392..2822
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     polyA_signal    2944..3065
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2981..3000)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3035..3054
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3072..3527)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3209..3230)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3421..3440
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3554..3658
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3660..4017
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3660..3851
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3683..3703)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3868..4003
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4052..4846
                     /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             4052..4846
                     /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(4106..4125)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4716..4735
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5034..5053)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5078..5125
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5454..6042
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5943..5962
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcatgg
      541 cagtgaatgt gtactctacg tcagtcacca gtgataacct aagtcgacat gacatgctgg
      601 cttggatcaa tgaatctctg cagttgaatc tgacaaagat agaacagttg tgttcagggg
      661 ctgcatattg tcagtttatg gacatgctct tccctggatc cattgccttg aagaaagtga
      721 aattccaagc taagctagaa catgaatata tccagaactt caaaatacta caagcaggct
      781 tcaagaggat gggcgttgac aaaataattc ctgtggataa attagtaaaa ggaaaatttc
      841 aggacaattt tgaatttgtt caatggttca agaagttttt tgatgcaaat tatgatggaa
      901 aagagtatga tcctgtagct gccagacaag gtcaagaaac tgcagtggct ccttctcttg
      961 tcgccccggc tttgagtaaa ccgaagaaac ctctcggctc cagtactgca gccccacaga
     1021 gacccattgc aacacagagg actactgcag ctcctaaggc tggccccgga atggtgcgaa
     1081 agaatcctgg tgtgggcaat ggagatgatg aagcagctga attgatgcag caggtcaaag
     1141 tactgaagct tactgttgaa gacttggaga aggagagaga cttctacttc ggaaagctaa
     1201 ggaacattga actgatttgc caggagaacg agggggaaaa cgaccctgta ctgcagagga
     1261 ttgtagatat tctttatgcc acagatgaag gctttgtgat acctgatgaa gggggcccac
     1321 aggaggaaca agaagagtat ggtggtggtg gtgctagggt accgcgggcc cgggatccac
     1381 cggtcgccac catggtgagc aagggcgagg aggtcatcaa agagttcatg cgcttcaagg
     1441 tgcgcatgga gggctccatg aacggccacg agttcgagat cgagggcgag ggcgagggcc
     1501 gcccctacga gggcacccag accgccaagc tgaaggtgac caagggcggc cccctgccct
     1561 tcgcctggga catcctgtcc ccccagttca tgtacggctc caaggcgtac gtgaagcacc
     1621 ccgccgacat ccccgattac aagaagctgt ccttccccga gggcttcaag tgggagcgcg
     1681 tgatgaactt cgaggacggc ggtctggtga ccgtgaccca ggactcctcc ctgcaggacg
     1741 gcacgctgat ctacaaggtg aagatgcgcg gcaccaactt cccccccgac ggccccgtaa
     1801 tgcagaagaa gaccatgggc tgggaggcct ccaccgagcg cctgtacccc cgcgacggcg
     1861 tgctgaaggg cgagatccac caggccctga agctgaagga cggcggccac tacctggtgg
     1921 agttcaagac catctacatg gccaagaagc ccgtgcaact gcccggctac tactacgtgg
     1981 acaccaagct ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgagc
     2041 gctccgaggg ccgccaccac ctgttcctgg ggcatggcac cggcagcacc ggcagcggca
     2101 gctccggcac cgcctcctcc gaggacaaca acatggccgt catcaaagag ttcatgcgct
     2161 tcaaggtgcg catggagggc tccatgaacg gccacgagtt cgagatcgag ggcgagggcg
     2221 agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggcggccccc
     2281 tgcccttcgc ctgggacatc ctgtcccccc agttcatgta cggctccaag gcgtacgtga
     2341 agcaccccgc cgacatcccc gattacaaga agctgtcctt ccccgagggc ttcaagtggg
     2401 agcgcgtgat gaacttcgag gacggcggtc tggtgaccgt gacccaggac tcctccctgc
     2461 aggacggcac gctgatctac aaggtgaaga tgcgcggcac caacttcccc cccgacggcc
     2521 ccgtaatgca gaagaagacc atgggctggg aggcctccac cgagcgcctg tacccccgcg
     2581 acggcgtgct gaagggcgag atccaccagg ccctgaagct gaaggacggc ggccactacc
     2641 tggtggagtt caagaccatc tacatggcca agaagcccgt gcaactgccc ggctactact
     2701 acgtggacac caagctggac atcacctccc acaacgagga ctacaccatc gtggaacagt
     2761 acgagcgctc cgagggccgc caccacctgt tcctgtacgg catggacgag ctgtacaagt
     2821 aggcggccgc gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct
     2881 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     2941 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     3001 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     3061 tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     3121 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     3181 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     3241 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     3301 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     3361 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     3421 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     3481 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt
     3541 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     3601 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc
     3661 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     3721 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     3781 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3841 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     3901 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     3961 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat
     4021 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     4081 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     4141 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     4201 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
     4261 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     4321 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     4381 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     4441 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     4501 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     4561 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     4621 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     4681 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     4741 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     4801 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     4861 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     4921 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     4981 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca
     5041 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     5101 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     5161 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     5221 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     5281 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa
     5341 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     5401 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     5461 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     5521 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     5581 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     5641 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     5701 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     5761 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     5821 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     5881 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     5941 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6001 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     6061 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     6121 cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt aatcaattac
     6181 ggggtcatta gttcatagcc catatatgga gttccg
//
