LOCUS       sequence-246426-        7939 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7939)
  AUTHORS   Lees JA, Saito M, Vidal M, Valentine M, Look T, Harlow E, Dyson N, 
            Helin K
  TITLE     The retinoblastoma protein binds to a family of E2F transcription 
            factors.
  JOURNAL   Mol Cell Biol. 1993 Dec;13(12):7813-25.
  PUBMED    8246996
REFERENCE   2  (bases 1 to 7939)
  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: "Mol Cell 
            Biol."; date: "1993-12"; volume: "13(12)"; pages: "7813-25"
COMMENT     Sequence Label: pCMV-Neo-Bam E2F3a
FEATURES             Location/Qualifiers
     source          1..7939
                     /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     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          696..1268
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(2861..2880)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2926..2981
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2980..2999)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3339..3355)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3339..3355)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3352..3374)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3363..3379
                     /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(3387..3393,3394..3411,3412..3417))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3432..3453
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3570..3587)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3741..4329)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3821..3840)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4500..5291,5292..5360))
                     /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     5123..5142
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5361..5465)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5533..5551
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5589..5611)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5711..5730
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5924..5946
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5938..5955
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5939..5955
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6366..6511
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             6568..7362
                     /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(6622..6641)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7232..7251
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7556..7575)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7600..7647
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
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 agctcgttta gtgaaccgtc
      601 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
      661 ccagcctccg gtcgaccgat cctgagaact tcagggtgag tttggggacc cttgattgtt
      721 ctttcttttt cgctattgta aaattcatgt tatatggagg gggcaaagtt ttcagggtgt
      781 tgtttagaat gggaagatgt cccttgtatc accatggacc ctcatgataa ttttgtttct
      841 ttcactttct actctgttga caaccattgt ctcctcttat tttcttttca ttttctgtaa
      901 ctttttcgtt aaactttagc ttgcatttgt aacgaatttt taaattcact tttgtttatt
      961 tgtcagattg taagtacttt ctctaatcac ttttttttca aggcaatcag ggtatattat
     1021 attgtacttc agcacagttt tagagaacaa ttgttataat taaatgataa ggtagaatat
     1081 ttctgcatat aaattctggc tggcgtggaa atattcttat tggtagaaac aactacaccc
     1141 tggtcatcat cctgcctttc tctttatggt tacaatgata tacactgttt gagatgagga
     1201 taaaatactc tgagtccaaa ccgggcccct ctgctaacca tgttcatgcc ttcttctctt
     1261 tcctacagct cctgggcaac gtgctggttg ttgtgctgtc tcatcatttt ggcaaagaat
     1321 tcctcgacgg atccatggtg agaaagggaa tccagcccgc tctggagcag tacctggtga
     1381 ccgccggggg tggggagggg gcggctgtcg tcgccgccgc cgctgcagcc tccatggaca
     1441 aaagggcact gctagccagc cccggcttcg ccgccgccgc cgccgctgcc gccgccccgg
     1501 gcgcgtacat ccagatcctc accacgaaca cttccaccac ctcctgttcc tcctccctcc
     1561 aaagcggcgc cgtagccgcc ggccccctcc tccccagtgc ccccggcgcg gagcagaccg
     1621 ccggcagcct cctctacacc acgccgcacg gaccctccag cagagccggg ctgctgcagc
     1681 agccaccagc gctgggacgc ggcggcagcg gcggcggcgg cggccctccg gcaaagcgaa
     1741 ggctggagct aggagaaagc ggtcatcagt acctctcaga tggtttaaaa acccccaagg
     1801 gcaaaggaag agctgcacta cgaagtccag atagtccaaa aactccaaaa tctccctcag
     1861 aaaaaacgcg gtatgatacg tctcttggtc tgctcaccaa gaagttcatt cagctcctga
     1921 gccagtcacc cgatggggta ttggatttga acaaggcagc agaagtgcta aaagtgcaaa
     1981 agagaaggat ttatgatatc accaacgttc tggaaggcat ccacctcatt aagaagaagt
     2041 ctaaaaacaa cgtccaatgg atgggctgca gtctgtctga ggatgggggc atgctggccc
     2101 agtgtcaagg cctgtcaaaa gaagtgaccg agctcagtca ggaagagaag aaattagatg
     2161 aactgatcca aagctgcacc ctggacctca aactgttaac cgaggattca gagaatcaaa
     2221 ggttagctta tgttacatat caagatattc gaaaaattag tggccttaaa gaccaaactg
     2281 ttatagttgt gaaagcccct ccagaaacaa gacttgaagt gcctgactca atagagagcc
     2341 tacaaataca tttggcaagt acccaagggc ccattgaggt ttacttatgt ccagaagaga
     2401 ctgaaacaca cagtccaatg aaaacaaaca accaagacca caatgggaat atccctaaac
     2461 ccgcttccaa agacttggct tcaaccaact caggacatag cgattgctca gtttctatgg
     2521 gaaacctttc tcctctggcc tccccagcca acctcttaca gcagactgag gaccaaattc
     2581 cttccaacct agaaggaccg tttgtgaact tactgcctcc cctgctgcaa gaggactatc
     2641 tcctgagcct cggggaggag gaaggcatca gcgatctctt cgatgcttac gatttggaaa
     2701 agctcccact ggtggaagac ttcatgtgta gttgattatg cttcgtgtga actctccttc
     2761 gatccggatc cgtcgaggaa ttcactcctc aggtgcaggc tgcctatcag aaggtggtgg
     2821 ctggtgtggc caatgccctg gctcacaaat accactgaga tctttttccc tctgccaaaa
     2881 attatgggga catcatgaag ccccttgagc atctgacttc tggctaataa aggaaattta
     2941 ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga
     3001 gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca acatatgccc
     3061 atatgctggc tgccatgaac aaaggttggc tataaagagg tcatcagtat atgaaacagc
     3121 cccctgctgt ccattcctta ttccatagaa aagccttgac ttgaggttag atttttttta
     3181 tattttgttt tgtgttattt ttttctttaa catccctaaa attttcctta catgttttac
     3241 tagccagatt tttcctcctc tcctgactac tcccagtcat agctgtccct cttctcttat
     3301 ggagatccct cgacggatcg gccgcaattc gtaatcatgt catagctgtt tcctgtgtga
     3361 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     3421 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     3481 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     3541 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     3601 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     3661 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3721 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3781 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3841 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3901 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3961 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4021 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4081 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     4141 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     4201 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     4261 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     4321 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     4381 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     4441 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     4501 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     4561 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     4621 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     4681 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     4741 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     4801 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     4861 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     4921 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     4981 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     5041 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     5101 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     5161 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     5221 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     5281 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     5341 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     5401 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     5461 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     5521 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac
     5581 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     5641 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
     5701 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata
     5761 ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc
     5821 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
     5881 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
     5941 aaaacgacgg ccagtgccaa gcttcacgct gccgcaagca ctcagggcgc aagggctgct
     6001 aaaggaagcg gaacacgtag aaagccagtc cgcagaaacg gtgctgaccc cggatgaatg
     6061 tcagctactg ggctatctgg acaagggaaa acgcaagcgc aaagagaaag caggtagctt
     6121 gcagtgggct tacatggcga tagctagact gggcggtttt atggacagca agcgaaccgg
     6181 aattgccagc tggggcgccc tctggtaagg ttgggaagcc ctgcaaagta aactggatgg
     6241 ctttcttgcc gccaaggatc tgatggcgca ggggatcaag atcctgcttc atccccgtgg
     6301 cccgttgctc gcgtttgctg gcggtgtccc cggaagaaat atatttgcat gtctttagtt
     6361 ctatgatgac acaaaccccg cccagcgtct tgtcattggc gaattcgaac acgcagatgc
     6421 agtcggggcg gcgcggtccc aggtccactt cgcatattaa ggtgacgcgt gtggcctcga
     6481 acaccgagcg accctgcagc gacccgctta acagcgtcaa cagcgtgccg cagatctgat
     6541 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     6601 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     6661 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     6721 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     6781 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     6841 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     6901 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     6961 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     7021 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     7081 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     7141 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     7201 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     7261 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     7321 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     7381 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     7441 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     7501 gcggggatct catgctggag ttcttcgccc accccgggag atgggggagg ctaactgaaa
     7561 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     7621 aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     7681 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     7741 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcaa
     7801 gccctgccat agccacgggc cccgtgggtt agggacggcg gatcgcggcc ctctagagag
     7861 cttggcccat tgcatacgtt gtatccatat cataatatgt acatttatat tggctcatgt
     7921 ccaacattac cgccatgtt
//
