LOCUS       sequence-424514-        7608 bp    DNA     circular SYN 14-APR-2025
DEFINITION  expression of GFP-tagged wild-type dynamitin p50 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7608)
  AUTHORS   Quintyne NJ, Gill SR, Eckley DM, Crego CL, Compton DA, Schroer TA
  TITLE     Dynactin is required for microtubule anchoring at centrosomes.
  JOURNAL   J Cell Biol. 1999 Oct 18;147(2):321-34.
  PUBMED    10525538
REFERENCE   2  (bases 1 to 7608)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Cell 
            Biol."; date: "1999-10-18"; volume: "147(2)"; pages: "321-34"
COMMENT     Sequence Label: Dynamitin-GFP
FEATURES             Location/Qualifiers
     source          1..7608
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     40..59
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        40..58
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             90..119
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             129..158
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             168..197
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             207..236
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             246..275
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             309..338
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             351..1064
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFAYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             351..1064
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFAYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(381..409)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1004..1024
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     promoter        complement(2338..2356)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2339..2356)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2376..2393)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2382..2606
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2652..3080
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2739..2758)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2949..2970
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3089..3109)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3094..3423
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3274..3409
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3336..3355
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3490..4284
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3544..3563)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4154..4173
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4458..4579
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4495..4514)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4549..4568
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4628..4644)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4628..4644)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4641..4663)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4652..4668
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4676..4682,4683..4700,4701..4706))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4721..4742
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4859..4876)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5030..5618)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5110..5129)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5789..6580,6581..6649))
                     /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     6412..6431
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6650..6754)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6829..6848)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        7020..7399
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7400..7603
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7554..7574
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc actataggga
       61 gacccaagct tcatcgattt aaagctatgg agcaaaagct catttctgaa gaggacttga
      121 atgaaatgga gcaaaagctc atttctgaag aggacttgaa tgaaatggag caaaagctca
      181 tttctgaaga ggacttgaat gaaatggagc aaaagctcat ttctgaagag gacttgaatg
      241 aaatggagca aaagctcatt tctgaagagg acttgaatga aatggagagc ttgggcgacc
      301 tcaccatgga gcaaaagctc atttctgaag aggacttgaa ttcctctaga atgagtaaag
      361 gagaagaact tttcactgga gttgtcccaa ttcttgttga attagatggt gatgttaatg
      421 ggcacaaatt ttctgtcagt ggagagggtg aaggtgatgc aacatacgga aaacttaccc
      481 ttaaatttat ttgcactact ggaaaactac ctgttccatg gccaacactt gtcactactt
      541 tcgcttatgg tgttcaatgc ttttcaagat acccagatca tatgaaacgg catgactttt
      601 tcaagagtgc catgcccgaa ggttatgtac aggaaagaac tatatttttc aaagatgacg
      661 ggaactacaa gacacgtgct gaagtcaagt ttgaaggtga tacccttgtt aatagaatcg
      721 agttaaaagg tattgatttt aaagaagatg gaaacattct tggacacaaa ttggaataca
      781 actataactc acacaatgta tacatcatgg cagacaaaca aaagaatgga atcaaagtta
      841 acttcaaaat tagacacaac attgaagatg gaagcgttca actagcagac cattatcaac
      901 aaaatactcc aattggcgat ggccctgtcc ttttaccaga caaccattac ctgtccacac
      961 aatctgccct ttcgaaagat cccaacgaaa agagagacca catggtcctt cttgagtttg
     1021 taacagctgc tgggattaca catggcatgg atgaactata caaaggatcc atggccgacc
     1081 ccaaatacgc ggacctgccc ggcatcgccc ggaatgagcc cgacgtgtac gagaccagcg
     1141 acctccccga ggatgaccag gccgagttcg aggcggagct ggaggagctc accagcacca
     1201 gcgtggagca cctcatcatc aaccccaacg ccgccttcga gaagttcaag gacaaacgcc
     1261 tgggcaccga tggcgtggat ttctccgacc gcatcagcaa gagcagaacc actggctatg
     1321 agtctgggga gtatgaaatt ctgggagagg ggttgggtgc caaggagacc ccccagcagc
     1381 ggtaccagcg gctgcagcat gaggtgcagg agctgatccg tgatgtggag cagatccaga
     1441 gcgccgtgaa ggagtcggcg gccgaggaag agctgacgcc catggctttg gcccggcagc
     1501 tggaggggct gaagcagcag ttggtttcgt gccatctgca gaagctgttg ggccccactg
     1561 ccgccatcga cttcgcggac cctgagggtg ccttggcgaa gcgcctgcag cagcagctgg
     1621 aagtagccaa gtgcaagaag gcagcgccag caaagagccc ccccaaagcc cctgcgccca
     1681 ccactgacgc gctcaccttc gaactgttct ggaggccaga gcaggaccag ttctcccaga
     1741 ctgccaagat agcagagctg gagaagcgcc tggcacagct ggaggccatg gtgcgctgtg
     1801 agcctgacag ccagaatcct ctgctagttg ggctgaaggg caccagcctc gtggagaccg
     1861 tgcagatcct gcaggccaag gttaacatcc tggacgcagc tgtgctggac caagtggagg
     1921 cccggctgca gagcgtcctg gggaaggtga atgagattgc caagcacaaa gcgattgtgc
     1981 aggatgccga cacgcagagc aagatccatc aggtctacga gatgatgcag cgctgggacc
     2041 acatggccag cagtctgccc gacgtggtgc agcggctgct gacgctccgg gacctacatg
     2101 agcaggcctc acgatttgtg caggtgctgg tgcacctgga caccacacag caggaggtgg
     2161 ctgctgctct gaaggataac actgtgctgc tggcagaggt ccagaagaca atgaaggaga
     2221 acctggctgt tgtagaggac aactttgctg aagtagaggc ccgcatcaag cggctgcagt
     2281 gatagcggcc ggtgatcact agtgcggccg ctcgagcatg catctagagg gccctattct
     2341 atagtgtcac ctaaatgcta gagctcgctg atcagcctcg actgtgcctt ctagttgcca
     2401 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     2461 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     2521 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     2581 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     2641 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     2701 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     2761 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     2821 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     2881 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     2941 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     3001 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     3061 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     3121 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     3181 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3241 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     3301 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     3361 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     3421 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     3481 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     3541 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     3601 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     3661 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     3721 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     3781 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     3841 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     3901 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     3961 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     4021 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     4081 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     4141 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     4201 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     4261 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     4321 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     4381 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     4441 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4501 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     4561 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     4621 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     4681 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     4741 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     4801 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     4861 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4921 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4981 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5041 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5101 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     5161 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     5221 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5281 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5341 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5401 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5461 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5521 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     5581 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     5641 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5701 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     5761 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     5821 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     5881 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     5941 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     6001 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     6061 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     6121 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     6181 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     6241 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     6301 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     6361 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6421 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6481 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     6541 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     6601 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     6661 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     6721 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     6781 acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc
     6841 tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta
     6901 gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa
     6961 tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg
     7021 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
     7081 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     7141 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     7201 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     7261 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     7321 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     7381 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
     7441 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
     7501 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
     7561 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctg
//
