LOCUS       sequence-169095-        5482 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 5482)
  AUTHORS   Sellers WR, Novitch BG, Miyake S, Heith A, Otterson GA, Kaye FJ, 
            Lassar AB, Kaelin WG
  TITLE     Stable binding to E2F is not required for the retinoblastoma protein
            to activate transcription, promote differentiation, and suppress 
            tumor cell growth.
  JOURNAL   Genes Dev. 1998 Jan 1. 12(1):95-106.
  PUBMED    9420334
REFERENCE   2  (bases 1 to 5482)
  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: "Genes Dev. 
            1998 Jan 1. 12(1):95-106."
COMMENT     Sequence Label: 408 pSG5L HA E2F1
FEATURES             Location/Qualifiers
     source          1..5482
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        76..286
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        98..294
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      145..280
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          325..897
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     905..924
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     952..971
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        952..970
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1004..1030
                     /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             1004..1030
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1010..1036
                     /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"
     polyA_signal    2419..2540
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2456..2475)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2510..2529
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2574..2591)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2745..3333)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2825..2844)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3504..4295,4296..4364))
                     /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     4127..4146
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4365..4469)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4537..4555
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4593..4615)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4715..4734
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(4801..5256)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4938..4959)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5150..5169
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5382..5404
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5396..5413
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5397..5413
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5450..5470)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaggaaag
       61 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
      121 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
      181 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
      241 gagctattcc agaagtagtg aagaggcttt tttggaggcc taggcttttg caaaaagctc
      301 cggatcgatc ctgagaactt cagggtgagt ttggggaccc ttgattgttc tttctttttc
      361 gctattgtaa aattcatgtt atatggaggg ggcaaagttt tcagggtgtt gtttagaatg
      421 ggaagatgtc ccttgtatca ccatggaccc tcatgataat tttgtttctt tcactttcta
      481 ctctgttgac aaccattgtc tcctcttatt ttcttttcat tttctgtaac tttttcgtta
      541 aactttagct tgcatttgta acgaattttt aaattcactt ttgtttattt gtcagattgt
      601 aagtactttc tctaatcact tttttttcaa ggcaatcagg gtatattata ttgtacttca
      661 gcacagtttt agagaacaat tgttataatt aaatgataag gtagaatatt tctgcatata
      721 aattctggct ggcgtggaaa tattcttatt ggtagaaaca actacatcct ggtcatcatc
      781 ctgcctttct ctttatggtt acaatgatat acactgtttg agatgaggat aaaatactct
      841 gagtccaaac cgggcccctc tgctaaccat gttcatgcct tcttcttttt cctacagctc
      901 ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt gtaatacgac
      961 tcactatagg gcgaattgtc gacccgggaa gcttaccatg gcctacccct acgacgtgcc
     1021 cgactacgcc tccctcggat cccatatggc cttggccggg gcccctgcgg gcggcccatg
     1081 cgcgccggcg ctggaggccc tgctcggggc cggcgcgctg cggctgctcg actcctcgca
     1141 gatcgtcatc atctccgccg cgcaggacgc cagcgccccg ccggctccca ccggccccgc
     1201 ggcgcccgcc gccggcccct gcgaccctga cctgctgctc ttcgccacac cgcaggcgcc
     1261 ccggcccaca cccagtgcgc cgcggcccgc gctcggccgc ccgccggtga agcggaggct
     1321 ggacctggaa actgaccatc agtacctggc cgagagcagt gggccagctc ggggcagagg
     1381 ccgccatcca ggaaaaggtg tgaaatcccc gggggagaag tcacgctatg agacctcact
     1441 gaatctgacc accaagcgct tcctggagct gctgagccac tcggctgacg gtgtcgtcga
     1501 cctgaactgg gctgccgagg tgctgaaggt gcagaagcgg cgcatctatg acatcaccaa
     1561 cgtccttgag ggcatccagc tcattgccaa gaagtccaag aaccacatcc agtggctggg
     1621 cagccacacc acagtgggcg tcggcggacg gcttgagggg ttgacccagg acctccgaca
     1681 gctgcaggag agcgagcagc agctggacca cctgatgaat atctgtacta cgcagctgcg
     1741 cctgctctcc gaggacactg acagccagcg cctggcctac gtgacgtgtc aggaccttcg
     1801 tagcattgca gaccctgcag agcagatggt tatggtgatc aaagcccctc ctgagaccca
     1861 gctccaagcc gtggactctt cggagaactt tcagatctcc cttaagagca aacaaggccc
     1921 gatcgatgtt ttcctgtgcc ctgaggagac cgtaggtggg atcagccctg ggaagacccc
     1981 atcccaggag gtcacttctg aggaggagaa cagggccact gactctgcca ccatagtgtc
     2041 accaccacca tcatctcccc cctcatccct caccacagat cccagccagt ctctactcag
     2101 cctggagcaa gaaccgctgt tgtcccggat gggcagcctg cgggctcccg tggacgagga
     2161 ccgcctgtcc ccgctggtgg cggccgactc gctcctggag catgtgcggg aggacttctc
     2221 cggcctcctc cctgaggagt tcatcagcct ttccccaccc cacgaggccc tcgactacca
     2281 cttcggcctc gaggagggcg agggcatcag agacctcttc gactgtgact ttggggacct
     2341 cacccccctg gatttctgac aggatgcatg aattccccga tatcgcctcg agtctagatc
     2401 cagatcttat taaagcagaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     2461 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     2521 aaactcatca atgtatctta tcatgtctgg tcgactctag actcttccgc ttcctcgctc
     2581 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     2641 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     2701 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     2761 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     2821 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     2881 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     2941 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     3001 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     3061 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     3121 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     3181 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     3241 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     3301 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     3361 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     3421 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     3481 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     3541 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     3601 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     3661 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     3721 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     3781 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     3841 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     3901 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     3961 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     4021 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     4081 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     4141 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     4201 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     4261 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     4321 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     4381 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     4441 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     4501 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     4561 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     4621 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     4681 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
     4741 gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     4801 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     4861 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     4921 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     4981 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     5041 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     5101 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     5161 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     5221 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccat tcgccattca ggctgcgcaa
     5281 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     5341 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
     5401 aacgacggcc agtgaattgt cgacttctga ggcggaaaga accagctgtg gaatgtgtgt
     5461 cagttagggt gtggaaagtc cc
//
