LOCUS       sequence-201686-        6731 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human gamma-tubulin with a GFP-tag on the C-terminal of 
            the protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6731)
  AUTHORS   Alvarado-Kristensson M, Rodriguez MJ, Silio V, Valpuesta JM, Carrera
            AC
  TITLE     SADB phosphorylation of gamma-tubulin regulates centrosome 
            duplication.
  JOURNAL   Nat Cell Biol. 2009 Sep;11(9):1081-92. doi: 10.1038/ncb1921. Epub 
            2009 Aug 2.
  PUBMED    19648910
REFERENCE   2  (bases 1 to 6731)
  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"; doi: "10.1038/ncb1921"; 
            journalName: "Nat Cell Biol"; date: "2009-09"; volume: "11"; issue: 
            "9"; pages: "1081-92"
COMMENT     Sequence Label: pEGFPN1-gamma-tubulin
FEATURES             Location/Qualifiers
     source          1..6731
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        259..562
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        563..766
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     717..737
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1824..1833
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1830..1832,1833..1835,1836..2549)
                     /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(1830..1832,1833..1835,1836..2546)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1875..1896)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2136..2155)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2483..2504
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2672..2793
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2709..2728)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2763..2782
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2800..3255)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2937..2958)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3149..3168
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3282..3386
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3388..3745
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3388..3579
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3411..3431)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3596..3731
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3780..4574
                     /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             3780..4574
                     /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(3834..3853)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4444..4463
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4762..4781)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4806..4853
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5182..5770
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5671..5690
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6054..6075
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6090..6095,6096..6113,6114..6120)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6128..6144
                     /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)."
     primer_bind     6133..6155
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6152..6168
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6152..6168
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ttgnatchcc twgagyaaam tcgcvgtdth aamtkccctg atgccacgta gttaagscag
       61 tatctggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
      121 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
      181 cgtattaccg ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag
      241 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
      301 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
      361 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
      421 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
      481 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
      541 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
      601 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
      661 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
      721 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg
      781 tcagatccgc tagcgatgcc gagggaaatc atcaccctac agttgggcca gtgcggcaat
      841 cagattgggt tcgagttctg gaaacagctg tgcgccgagc atggtatcag ccccgagggc
      901 atcgtggagg agttcgccac cgagggcact gaccacaagg acgtcttttt ctaccaggca
      961 gacgatgagc actacatccc ccgggccgtg ctgctggact tggaaccccg ggtgatccac
     1021 tccatcctca actcccccta tgccaagctc tacaacccag agaacatcta cctgtcggaa
     1081 catggaggag gagctggcaa caactgggcc agcggattct cccagggaga aaagatccat
     1141 gaggacattt ttgacatcat agaccgggag gcagatggta gtgacagtct agagggcttt
     1201 gtgctgtgtc actccattgc tggggggaca ggctctggac tgggttccta cctcttagaa
     1261 cggctgaatg acaggtatcc taagaagctg gtgcagacat actcagtgtt tcccaaccag
     1321 gacgagatga gcgatgtggt ggtccagcct tacaattcac tcctcacact caagaggctg
     1381 acgcagaatg cagactgtgt ggtggtgctg gacaacacag ccctgaaccg gattgccaca
     1441 gaccgcctgc acatccagaa cccatccttc tcccagatca accagctggt gtctaccatc
     1501 atgtcagcca gcaccaccac cctgcgctac cctggctaca tgaacaatga cctcatcggc
     1561 ctcatcgcct cgctcattcc caccccacgg ctccacttcc tcatgaccgg ctacacccct
     1621 ctcactacgg accagtcagt ggccagcgtg aggaagacca cggtcctgga tgtcatgagg
     1681 cggctgctgc agcccaagaa cgtgatggtg tccacaggcc gagaccgcca gaccaaccac
     1741 tgctacatcg ccatcctcaa catcatccag ggagaggtgg accccaccca ggtccacaag
     1801 agcttgcaga ggatccaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg
     1861 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     1921 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     1981 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
     2041 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     2101 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     2161 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     2221 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     2281 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     2341 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     2401 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
     2461 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     2521 ctctcggcat ggacgagctg tacaagtaaa gcggccgcga ctctagatca taatcagcca
     2581 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     2641 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     2701 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     2761 ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt
     2821 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     2881 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     2941 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     3001 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     3061 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     3121 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     3181 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     3241 cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     3301 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     3361 ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg
     3421 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3481 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3541 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     3601 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     3661 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     3721 ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca
     3781 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     3841 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     3901 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     3961 aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     4021 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     4081 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     4141 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     4201 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     4261 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg
     4321 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     4381 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     4441 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     4501 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     4561 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     4621 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     4681 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     4741 ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc
     4801 tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg
     4861 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     4921 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     4981 gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact
     5041 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     5101 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     5161 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     5221 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     5281 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     5341 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     5401 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     5461 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     5521 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     5581 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     5641 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     5701 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     5761 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     5821 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat
     5881 gcattagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
     5941 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     6001 cattgacgtc aatggtatgc ckactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     6061 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     6121 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     6181 aagcttgcat gcctgcaggt cgacaagccc tttcaggtag agccagtacc tgttccccac
     6241 actgaaagct agggtgctgg gcagggtggt gatatcaagt accatggggc gcctgtgggg
     6301 gcatcttatt cccactccca cttccatagc tctctgaagg tctgcttggg ttcctggtta
     6361 cttactgcat gtgactgcag gtgttacata ctaggggatg ggagtatctg tcgggagaaa
     6421 cagggtaatg tgagtgagag aaccccgcac taaattccta gcatgtttac agagactcag
     6481 agattgtatg tttggtagtg agtagtgtgt tgaattactg agagagtgct cataagcact
     6541 gttggctaca tctccagggg acacttggtg ctgcttatag aaaaatcaag tgagtctgtg
     6601 acagttggca tcactaaata tcttaaatca tctgtattta ttctactttc tggagtgtct
     6661 ctgaaaatac tcaggcctta aatttgcact tagtacaacc cttctgtttt tgtaagagct
     6721 aaccatgtgt t
//
