LOCUS       sequence-458441-        6827 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TUBG2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6827)
  AUTHORS   Lindstrom L, Zhou J, Villoutreix BO, Malycheva D, Otrocka M, 
            Gustavsson AL, Lundback T, Bliman D, Shameem MA, Straw M, Riesbeck 
            K, Olsson R, Alvarado-Kristensson M
  TITLE     Targeting TUBG1 in RB1-negative tumors.
  JOURNAL   FASEB J. 2025 Mar 15;39(5):e70431. doi: 10.1096/fj.202403180RR.
  PUBMED    40019206
REFERENCE   2  (bases 1 to 6827)
  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: "FASEB J."; 
            date: "2025-03-15"; volume: "39(5):e70431. doi"; pages: " 
            10.1096/fj.202403180RR"
COMMENT     Sequence Label: pcDNA3.1 Flag-TUBG2
FEATURES             Location/Qualifiers
     source          1..6827
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     393..412
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(631..735)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(810..829)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1001..1380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1381..1584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1535..1555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1629..1648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1629..1647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1716..1739
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(3146..3164)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3147..3164)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3184..3201)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3190..3414
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3460..3888
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3547..3566)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3757..3778
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3897..3917)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3902..4231
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4082..4217
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4144..4163
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4298..5092
                     /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(4352..4371)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4962..4981
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5266..5387
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5303..5322)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5357..5376
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5436..5452)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5436..5452)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5449..5471)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5460..5476
                     /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(5484..5490,5491..5508,5509..5514))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5529..5550
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5667..5684)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5838..6426)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5918..5937)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6597..6827,1..561,562..630))
                     /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"
ORIGIN
        1 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
       61 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
      121 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
      181 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
      241 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
      301 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
      361 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
      421 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
      481 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
      541 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
      601 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
      661 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
      721 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
      781 tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
      841 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
      901 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt
      961 tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt
     1021 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     1081 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     1141 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     1201 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     1261 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     1321 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     1381 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
     1441 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
     1501 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
     1561 tgggaggtct atataagcag agctctctgg ctaactagag aacccactgc ttactggctt
     1621 atcgaaatta atacgactca ctatagggag acccaagctt ggtaccgagc tcggatccac
     1681 tagtaacggc cgccagtgtg ctggaattca ccatggatta caaggatgac gacgataaga
     1741 tcccccggga gatcatcacc ctgcagctag gacagtgcgg caaccagatt gggttcgagt
     1801 tctggaaaca gctgtgcgcc gagcatggta tcagccccga gggcatcgtg gaggagttcg
     1861 ccaccgaggg cactgaccgc aaggacgtct ttttctacca ggcagacgat gagcactaca
     1921 tcccccgggc cgtgctgctg gacttggaac cccgggtgat ccactccatc ctcaactccc
     1981 cctatgccaa gctctacaac ccagagaaca tctacctgtc ggaacatgga ggaggagctg
     2041 gcaacaactg ggccagcgga ttctcccagg gtgagaaaat tcatgaagac atctttgaca
     2101 tcatagaccg agaagcagat ggaagtgaca gtttggaggg cttcgtgctg tgtcactcca
     2161 tcgctggggg tacgggttct ggcctgggct cctacctcct ggagcgactg aatgacaggt
     2221 accccaagaa gctagtgcag acttattcag tgtttcccta ccaggacgag atgagcgacg
     2281 tagtggttca gccctacaat tcactcctga cactcaagag gctgacgcag aacgcagatt
     2341 gtgtggtggt gctggacaac acagccctga accggattgc cacagaccgc ctgcacatcc
     2401 agaacccgtc cttctcccag atcaaccagc tggtgtccac catcatgtcg gccagcacca
     2461 ccaccctgcg ctaccccggc tacatgaaca atgacctcat cggcctcatc gcctcgctca
     2521 ttcccacccc acggctccac ttcctcatga ccggctacac cccgctcact acagaccagt
     2581 cagtggccag cgtgaggaag accacggtcc tggatgtcat gaggcggctg ctgcagccca
     2641 agaacgtgat ggtgtccaca ggccgagacc gccagaccaa ccactgctac atcgccatcc
     2701 tcaacatcat ccagggagag gtggacccca cccaggtcca caagagcctg cagaggatcc
     2761 gggaacggaa gttggccaac ttcatcccgt ggggccccgc cagcatccag gtggccctgt
     2821 cgaggaagtc tccctacctg ccctcggccc accgggtcag cgggctcatg atggccaacc
     2881 acaccagcat ctcctcgctc tttgaaagtt cctgccagca gtttgacaag ctgcggaagc
     2941 gggatgcctt cctcgagcag ttccgtaagg aggatatgtt taaggacaac tttgatgaga
     3001 tggacaggtc tagggaggtt gttcaggagc tcattgatga gtaccatgcg gccacccagc
     3061 cagactacat ttcctggggc acccaggagc agtgagatat ccatcacact ggcggccgct
     3121 cgagcatgca tctagagggc cctattctat agtgtcacct aaatgctaga gctcgctgat
     3181 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     3241 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     3301 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     3361 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     3421 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     3481 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     3541 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3601 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     3661 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     3721 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     3781 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     3841 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     3901 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3961 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     4021 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4081 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4141 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     4201 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     4261 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     4321 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     4381 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     4441 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     4501 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     4561 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     4621 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     4681 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     4741 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     4801 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     4861 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     4921 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     4981 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     5041 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     5101 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     5161 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     5221 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     5281 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     5341 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     5401 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     5461 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     5521 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     5581 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     5641 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     5701 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     5761 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5821 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5881 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5941 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6001 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     6061 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6121 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6181 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6241 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     6301 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6361 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6421 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6481 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6541 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     6601 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     6661 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     6721 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     6781 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgc
//
