LOCUS       sequence-178205-        6088 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses human gamma-tubulin with a GFP-tag on the N-terminal of 
            the protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6088)
  AUTHORS   Rossello CA, Lindstrom L, Glindre J, Eklund G, Alvarado-Kristensson 
            M
  TITLE     Gamma-tubulin coordinates nuclear envelope assembly around 
            chromatin.
  JOURNAL   Heliyon. 2016 Sep 24;2(9):e00166. eCollection 2016 Sep.
  PUBMED    27699285
REFERENCE   2  (bases 1 to 6088)
  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: "Heliyon."; 
            date: "2016-09-24"; volume: "2(9)"; pages: "e00166. eCollection 2016
            Sep"
COMMENT     Sequence Label: TUBG1
FEATURES             Location/Qualifiers
     source          1..6088
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     575..594
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(893..912)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    937..984
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1313..1901
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1802..1821
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2076..2379
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2380..2583
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2534..2554
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2622..2631
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2628..2630,2631..2633,2634..3344)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2628..2630,2631..2633,2634..3344)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2673..2694)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2934..2953)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3281..3302
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4891..5012
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4928..4947)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4982..5001
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5019..5474)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5156..5177)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5368..5387
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5501..5605
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5607..5964
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5607..5798
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5630..5650)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5815..5950
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(5999..6088,1..705)
                     /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             join(5999..6088,1..705)
                     /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(6053..6072)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
       61 accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg
      121 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
      181 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
      241 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
      301 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
      361 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
      421 ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
      481 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
      541 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
      601 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
      661 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
      721 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
      781 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
      841 agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac
      901 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
      961 cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg
     1021 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac
     1081 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc
     1141 ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat
     1201 ttaaaagggt ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     1261 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     1321 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     1381 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     1441 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     1501 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     1561 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     1621 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     1681 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     1741 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     1801 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     1861 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     1921 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     1981 ctgattctgt ggataaccgt attaccgcca tgcattagtt attaatagta atcaattacg
     2041 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
     2101 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     2161 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     2221 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
     2281 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
     2341 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
     2401 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
     2461 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
     2521 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
     2581 gctggtttag tgaaccgtca gatccgctag cgctaccggt cgccaccatg gtgagcaagg
     2641 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     2701 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     2761 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     2821 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
     2881 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     2941 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     3001 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     3061 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
     3121 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
     3181 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
     3241 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     3301 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtccgga ctcagatctc
     3361 gagctcaagc ttcgatgccg agggaaatca tcaccctaca gttgggccag tgcggcaatc
     3421 agattgggtt cgagttctgg aaacagctgt gcgccgagca tggtatcagc cccgagggca
     3481 tcgtggagga gttcgccacc gagggcactg accacaagga cgtctttttc taccaggcag
     3541 acgatgagca ctacatcccc cgggccgtgc tgctggactt ggaaccccgg gtgatccact
     3601 ccatcctcaa ctccccctat gccaagctct acaacccaga gaacatctac ctgtcggaac
     3661 atggaggagg agctggcaac aactgggcca gcggattctc ccagggagaa aagatccatg
     3721 aggacatttt tgacatcata gaccgggagg cagatggtag tgacagtcta gagggctttg
     3781 tgctgtgtca ctccattgct ggggggacag gctctggact gggttcctac ctcttagaac
     3841 ggctgaatga caggtatcct aagaagctgg tgcagacata ctcagtgttt cccaaccagg
     3901 acgagatgag cgatgtggtg gtccagcctt acaattcact cctcacactc aagaggctga
     3961 cgcagaatgc agactgtgtg gtggtgctgg acaacacagc cctgaaccgg attgccacag
     4021 accgcctgca catccagaac ccatccttct cccagatcaa ccagctggtg tctaccatca
     4081 tgtcagccag caccaccacc ctgcgctacc ctggctacat gaacaatgac ctcatcggcc
     4141 tcatcgcctc gctcattccc accccacggc tccacttcct catgaccggc tacacccctc
     4201 tcactacgga ccagtcagtg gccagcgtga ggaagaccac ggtcctggat gtcatgaggc
     4261 ggctgctgca gcccaagaac gtgatggtgt ccacaggccg agaccgccag accaaccact
     4321 gctacatcgc catcctcaac atcatccagg gagaggtgga ccccacccag gtccacaaga
     4381 gcttgcagag gatccgggaa cgcaagttgg ccaacttcat cccgtggggc cccgccagca
     4441 tccaggtggc cctgtcgagg aagtctccct acctgccctc ggcccaccgg gtcagcgggc
     4501 tcatgatggc caaccacacc agcatctcct cgctcttcga gagaacctgt cgccagtatg
     4561 acaagctgcg taagcgggag gccttcctgg agcagttccg caaggaggat atgtttaagg
     4621 acaactttga tgagatggac acatccaggg agattgtgca gcagctcatc gatgagtacc
     4681 atgcggccac acggccagac tacatctcct ggggcaccca ggagcagtga gaattctgca
     4741 gtcgacggta ccgcgggccc gggatccacc ggatctagat aactgatcat aatcagccat
     4801 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     4861 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     4921 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     4981 tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg ttaatatttt
     5041 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     5101 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     5161 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     5221 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     5281 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     5341 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     5401 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     5461 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     5521 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     5581 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
     5641 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     5701 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     5761 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     5821 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     5881 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     5941 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
     6001 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     6061 ctatgactgg gcacaacaga caatcggc
//
