LOCUS       sequence-458442-        6948 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RB1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6948)
  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 6948)
  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: pSG5LHARB1sgresistant
FEATURES             Location/Qualifiers
     source          1..6948
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(260..456)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        complement(268..478)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      274..409
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     566..586
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     complement(623..640)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(623..639)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(632..654)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      780..1235
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(867..886)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1077..1098
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1302..1321)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1421..1443
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1481..1499)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1567..1671
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1672..1740,1741..2532)
                     /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     complement(1890..1909)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2703..3291
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3192..3211
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3445..3462
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    3496..3617
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3507..3526)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3561..3580
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(6466..6492)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     CDS             complement(6472..6498)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             complement(6472..6498)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(6531..6550)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6532..6550)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6578..6597)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          join(6605..6948,1..229)
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
ORIGIN
        1 gcaagctaaa gtttaacgaa aaagttacag aaaatgaaaa gaaaataaga ggagacaatg
       61 gttgtcaaca gagtagaaag tgaaagaaac aaaattatca tgagggtcca tggtgataca
      121 agggacatct tcccattcta aacaacaccc tgaaaacttt gccccctcca tataacatga
      181 attttacaat agcgaaaaag aaagaacaat caagggtccc caaactcacc ctgaagttct
      241 caggatcgat ccggagcttt ttgcaaaagc ctaggcctcc aaaaaagcct cttcactact
      301 tctggaatag ctcagaggcc gaggcggcct cggcctctgc ataaataaaa aaaattagtc
      361 agccatgggg cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta
      421 ggggcgggac tatggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg
      481 gggagcctgg ggactttcct tgctgactaa ttgagatgca tgctttgcat acttctgcct
      541 gctggggagc ctggggactt tccacaccct aactgacaca cattccacag ctggttcttt
      601 ccgcctcaga agtcgacaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
      661 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
      721 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgga
      781 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
      841 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
      901 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
      961 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     1021 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     1081 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     1141 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     1201 cgcgaatttt aacaaaatat taacgcttac aatttcctga tgcggtattt tctccttacg
     1261 catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc
     1321 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     1381 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     1441 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     1501 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     1561 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     1621 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     1681 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     1741 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     1801 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     1861 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     1921 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     1981 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     2041 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     2101 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     2161 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     2221 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     2281 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     2341 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     2401 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     2461 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     2521 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     2581 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     2641 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     2701 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     2761 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     2821 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     2881 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     2941 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     3001 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     3061 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     3121 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     3181 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     3241 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     3301 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     3361 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     3421 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagtctaga
     3481 gtcgaccaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     3541 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     3601 gctgcaataa acaagttctg ctttaataag atctggatct agactcgagg cgatatcggg
     3661 gaattctcat ttctcttcct tgtttgaggt atccatgcta tcattcattt tctgcttttg
     3721 cattcgtgtt cgagtagaag tcatttctgc cagtttctgc tgaaatttgg actctcctgg
     3781 gagatgttta cttccatctg cttcatctga tccttcaata tcaaagcgta gttttttcag
     3841 tggtttagga gggttgcttc cttcagcact tcttttgagc acacggtcgc tgttacatac
     3901 catctgattt attttctgga acttctcaga agtcccgaat gattcaccaa ttgatactaa
     3961 gattcttgat cttggagtca tttttgttgg tgttggcaga ccttctgaaa ttttatatgg
     4021 actcttcagg ggtgaaatat agatgttccc tccaggaatc cgtaagggtg aactaggaaa
     4081 cttgtaaggg cttcgaggaa tgtgaggtat tggtgacaag gtagggggcc tggtggaagc
     4141 atactgcaaa atatttgttt tcagtctctg catgaagacc gagttataga atactataat
     4201 agaatcatac tcctcttctt tgatcaaaac acgtttgaat gtctcctgaa cagcatgagg
     4261 aagatccttg tatgctgtta caatgatttt gaatttaagg tctatattct tcactttgca
     4321 tatgccatac atggaacaca tcataatttg gtccaaatgc ctgtctctca tgagttcata
     4381 ctcattctgc agggtgtgct ggaaaagggt ccagatgata tgttctaatt ctgggtgctc
     4441 agacagaagg cgttcacaaa gtgtatttag ccggagatag gctagccgat acactttttt
     4501 ataaaacagt gaaagagagg tagatttcaa tggcttctgg gtctggaagg ctgaggttgc
     4561 ttgtgtctct gcatttgcag tagaatttac acgcgtagtt gaaccttttt tctttggaga
     4621 tcttacagga gaaagataca tatctgctgc agtgtgatta ttctggagag gaagattaag
     4681 aggacaagca gattcaaggt gatcagttgg tccttctcgg tcctttgatt gtttaataag
     4741 atcaaataaa ggtgaatctg agagccatgc aagggattcc atgattcgat gttcacatcg
     4801 ttctaaatgt tttatcattt ctcttgtcaa gttgccttct gctttgataa aactttcgat
     4861 cactttgtaa aaatcaaagg cttttaaatt aagcacattc agaatccatg ggaaagacaa
     4921 atctgttcca gaatcaagat tctgagatgt acttctgcta tatgtggcca ttacaacctc
     4981 aagagcgcac gccaataaag acatatgaaa aatgttgtca ttcagaagtt tgctaaaatt
     5041 ttgaatggat aatcgttctt cttctgattt aagcatggat tccattactc ggtaatacaa
     5101 gcgaactcca agtttgtatc gctgtgatcc aatttcgaca caaccctgtc ccacagcttt
     5161 agcaaatttc tctttaaaga tgtatcctat atccttcact cttttcagta tactttcttt
     5221 tggattcact gtgcagttgt taaaatagga aatcagattt tctgaaggtt gatcacttgc
     5281 tgaatttaaa atcatcatta attgttggat agtgttcata acagtcctaa ctggagtgtg
     5341 tggaggaatt acattcacct cttcatcaag gttacttttt cgtggtgttc tctgtgtttc
     5401 aaaactgtct atagaatcag tctgaagagt tttatcatga tccaaaaata atcttgcatc
     5461 tagatcttta tttttaagat aaatttcttc gtatcgttta gaaagatttt caacctctgg
     5521 aagtccatta gatgttacaa gtccaagaga attcataaaa ggtataaaat ttttgaaata
     5581 aacatttttc acctcatcta tattacattc atgttcttta cagagaactt caataattct
     5641 tgtatcattt tctagttgtt ttgctatccg tgcactcctg ttctgacctc gcctgggtgt
     5701 tcgaggtgaa ccattaatgg gtataacagc tgttttatat ggttctttga gcaacatggg
     5761 aggtgagagt ttaataaaat agtcaaggac acatagcatt aactgaaatg aaatcaccag
     5821 atcatcttcc atttgtaata cttccccttt agctaataaa aatgtgatcc aagaaacttt
     5881 tagcaccaat gcagaattta tttcagtaga tatcgaactg ctgggttgtg tcaaatatat
     5941 aagttcacat gtcctttcca atttgctgaa gagtgcaaac aatacatcat acttcttcaa
     6001 cagtcttgac atagcattat caactttggt actggtatca atttctttta gtaagttaaa
     6061 gaatttatgg acactgattt ctatgttttt ctgtagctca gtaaaagtga acgacatctc
     6121 atctaggtca actgctgcaa taaagataca gattccccac agttcctttt tcttttgaat
     6181 ataacctccc aatactccat ccacagatga aactttctcc caagttaacc aagctctctc
     6241 tctgacatga tctggtatct ttaatttctg acataatgca gtaaaatcag gttcttctgt
     6301 ttcttcaaac tcaagcctga cgagaggcag gtcctccggg ccgctgtcct gctctgggtc
     6361 ctcctcagga gggggcggcg gcggcggtgc cgggggttcc gcggcggcag cggcggcggt
     6421 ggccgcggtc tttcgggggg ttttgggcgg catggtggcg gatccgaggg aggcgtagtc
     6481 gggcacgtcg taggggtagg ccatggtaag cttcccgggt cgacaattcg ccctatagtg
     6541 agtcgtatta caattctttg ccaaaatgat gagacagcac aataaccagc acgttgccca
     6601 ggagctgtag gaaaaagaag aaggcatgaa catggttagc agaggggccc ggtttggact
     6661 cagagtattt tatcctcatc tcaaacagtg tatatcattg taaccataaa gagaaaggca
     6721 ggatgatgac caggatgtag ttgtttctac caataagaat atttccacgc cagccagaat
     6781 ttatatgcag aaatattcta ccttatcatt taattataac aattgttctc taaaactgtg
     6841 ctgaagtaca atataatata ccctgattgc cttgaaaaaa aagtgattag agaaagtact
     6901 tacaatctga caaataaaca aaagtgaatt taaaaattcg ttacaaat
//
