LOCUS       sequence-337480-        6800 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression in cell lines of the TUBG2 gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6800)
  AUTHORS   Corvaisier M, Zhou J, Malycheva D, Cornella N, Chioureas D, 
            Gustafsson NMS, Rossello CA, Ayora S, Li T, Ekstrom-Holka K, 
            Jirstrom K, Lindstrom L, Alvarado-Kristensson M
  TITLE     The gamma-tubulin meshwork assists in the recruitment of PCNA to 
            chromatin in mammalian cells.
  JOURNAL   Commun Biol. 2021 Jun 22;4(1):767. doi: 10.1038/s42003-021-02280-1.
  PUBMED    34158617
REFERENCE   2  (bases 1 to 6800)
  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: "Commun 
            Biol."; date: "2021-06-22"; volume: "4(1):767. doi"; pages: " 
            10.1038/s42003-021-02280-1"
COMMENT     Sequence Label: pcDNA3.1-TUBG2
FEATURES             Location/Qualifiers
     source          1..6800
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2119..2137)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2120..2137)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2157..2174)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2163..2387
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2433..2861
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2520..2539)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2730..2751
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2870..2890)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2875..3204
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3055..3190
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3117..3136
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3271..4065
                     /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(3325..3344)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3935..3954
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4239..4360
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4276..4295)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4330..4349
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4409..4425)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4409..4425)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4422..4444)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4433..4449
                     /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(4457..4463,4464..4481,4482..4487))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4502..4523
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4640..4657)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4811..5399)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4891..4910)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5570..6361,6362..6430))
                     /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     6193..6212
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6431..6535)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6610..6629)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ggtaccgagc tcggatccac tagtaacggc cgccagtgtg ctggaattca ccatgccccg
      721 ggagatcatc accctgcagc tgggccagtg cggcaaccag attgggttcg agttctggaa
      781 acagctgtgc gccgagcatg gtatcagccc cgagggcatc gtggaggagt tcgccaccga
      841 gggcactgac cgcaaggacg tctttttcta ccaggcagac gatgagcact acatcccccg
      901 ggccgtgctg ctggacttgg aaccccgggt gatccactcc atcctcaact ccccctatgc
      961 caagctctac aacccagaga acatctacct gtcggaacat ggaggaggag ctggcaacaa
     1021 ctgggccagc ggattctccc agggtgagaa aattcatgaa gacatctttg acatcataga
     1081 ccgagaagca gatggaagtg acagtttgga gggcttcgtg ctgtgtcact ccatcgctgg
     1141 gggtacgggt tctggcctgg gctcctacct cctggagcga ctgaatgaca ggtaccccaa
     1201 gaagctagtg cagacttatt cagtgtttcc ctaccaggac gagatgagcg acgtagtggt
     1261 tcagccctac aattcactcc tgacactcaa gaggctgacg cagaacgcag attgtgtggt
     1321 ggtgctggac aacacagccc tgaaccggat tgccacagac cgcctgcaca tccagaaccc
     1381 gtccttctcc cagatcaacc agctggtgtc caccatcatg tcggccagca ccaccaccct
     1441 gcgctacccc ggctacatga acaatgacct catcggcctc atcgcctcgc tcattcccac
     1501 cccacggctc cacttcctca tgaccggcta caccccgctc actacagacc agtcagtggc
     1561 cagcgtgagg aagaccacgg tcctggatgt catgaggcgg ctgctgcagc ccaagaacgt
     1621 gatggtgtcc acaggccgag accgccagac caaccactgc tacatcgcca tcctcaacat
     1681 catccaggga gaggtggacc ccacccaggt ccacaagagc ctgcagagga tccgggaacg
     1741 gaagttggcc aacttcatcc cgtggggccc cgccagcatc caggtggccc tgtcgaggaa
     1801 gtctccctac ctgccctcgg cccaccgggt cagcgggctc atgatggcca accacaccag
     1861 catctcctcg ctctttgaaa gttcctgcca gcagtttgac aagctgcgga agcgggatgc
     1921 cttcctcgag cagttccgta aggaggacat gttcaaggac aactttgatg agatggacag
     1981 gtctagggag gttgttcagg agctcattga tgagtaccat gcggccaccc agccagacta
     2041 catttcctgg ggcacccagg agcagtgaga tatccatcac actggcggcc gctcgagcat
     2101 gcatctagag ggccctattc tatagtgtca cctaaatgct agagctcgct gatcagcctc
     2161 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     2221 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     2281 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     2341 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     2401 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     2461 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     2521 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2581 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     2641 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     2701 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     2761 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     2821 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     2881 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     2941 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3001 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     3061 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     3121 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     3181 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     3241 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     3301 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     3361 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     3421 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     3481 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     3541 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     3601 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     3661 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     3721 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     3781 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     3841 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     3901 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     3961 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     4021 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     4081 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     4141 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     4201 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     4261 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4321 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     4381 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     4441 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     4501 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     4561 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     4621 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4681 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4741 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4801 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4861 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4921 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4981 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     5041 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5101 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     5161 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     5221 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     5281 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5341 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5401 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5461 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     5521 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     5581 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5641 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     5701 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     5761 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     5821 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     5881 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     5941 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6001 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6061 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     6121 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     6181 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     6241 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     6301 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     6361 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6421 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6481 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     6541 ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg
     6601 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg
     6661 tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt
     6721 gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt
     6781 acgggccaga tatacgcgtt
//
