LOCUS       sequence-166244-        6948 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 6948)
  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 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: "Genes Dev. 
            1998 Jan 1. 12(1):95-106."
COMMENT     Sequence Label: 413 pSG5L HA RB
FEATURES             Location/Qualifiers
     source          1..6948
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(11..28)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(941..1732,1733..1801))
                     /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     1564..1583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1802..1906)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1974..1992
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2030..2052)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2152..2171
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2238..2693)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2375..2396)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2587..2606
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2819..2841
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2833..2850
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2834..2850
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2887..2907)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2995..3205
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        3017..3213
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3064..3199
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          3244..3816
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     3824..3843
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     3871..3890
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3871..3889
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3923..3949
                     /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             3923..3949
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3929..3955
                     /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    6804..6925
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6841..6860)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6895..6914
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     1981 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     2041 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     2101 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     2161 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     2221 gaaaataccg catcaggaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     2281 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     2341 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     2401 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     2461 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     2521 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     2581 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     2641 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtccattcg
     2701 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     2761 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     2821 cagtcacgac gttgtaaaac gacggccagt gaattgtcga cttctgaggc ggaaagaacc
     2881 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     2941 gtatgcaaag catgcatctc aattagtcag caaggaaagt ccccaggctc cccagcaggc
     3001 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     3061 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     3121 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     3181 agaggctttt ttggaggcct aggcttttgc aaaaagctcc ggatcgatcc tgagaacttc
     3241 agggtgagtt tggggaccct tgattgttct ttctttttcg ctattgtaaa attcatgtta
     3301 tatggagggg gcaaagtttt cagggtgttg tttagaatgg gaagatgtcc cttgtatcac
     3361 catggaccct catgataatt ttgtttcttt cactttctac tctgttgaca accattgtct
     3421 cctcttattt tcttttcatt ttctgtaact ttttcgttaa actttagctt gcatttgtaa
     3481 cgaattttta aattcacttt tgtttatttg tcagattgta agtactttct ctaatcactt
     3541 ttttttcaag gcaatcaggg tatattatat tgtacttcag cacagtttta gagaacaatt
     3601 gttataatta aatgataagg tagaatattt ctgcatataa attctggctg gcgtggaaat
     3661 attcttattg gtagaaacaa ctacatcctg gtcatcatcc tgcctttctc tttatggtta
     3721 caatgatata cactgtttga gatgaggata aaatactctg agtccaaacc gggcccctct
     3781 gctaaccatg ttcatgcctt cttctttttc ctacagctcc tgggcaacgt gctggttatt
     3841 gtgctgtctc atcattttgg caaagaattg taatacgact cactataggg cgaattgtcg
     3901 acccgggaag cttaccatgg cctaccccta cgacgtgccc gactacgcct ccctcggatc
     3961 cgccaccatg ccgcccaaaa ccccccgaaa aacggccgcc accgccgccg ctgccgccgc
     4021 ggaacccccg gcaccgccgc cgccgccccc tcctgaggag gacccagagc aggacagcgg
     4081 cccggaggac ctgcctctcg tcaggcttga gtttgaagaa acagaagaac ctgattttac
     4141 tgcattatgt cagaaattaa agataccaga tcatgtcaga gagagagctt ggttaacttg
     4201 ggagaaagtt tcatctgtgg atggagtatt gggaggttat attcaaaaga aaaaggaact
     4261 gtggggaatc tgtatcttta ttgcagcagt tgacctagat gagatgtcgt tcacttttac
     4321 tgagctacag aaaaacatag aaatcagtgt ccataaattc tttaacttac taaaagaaat
     4381 tgataccagt accaaagttg ataatgctat gtcaagactg ttgaagaagt atgatgtatt
     4441 gtttgcactc ttcagcaaat tggaaaggac atgtgaactt atatatttga cacaacccag
     4501 cagttcgata tctactgaaa taaattctgc attggtgcta aaagtttctt ggatcacatt
     4561 tttattagct aaaggggaag tattacaaat ggaagatgat ctggtgattt catttcagtt
     4621 aatgctatgt gtccttgact attttattaa actctcacct cccatgttgc tcaaagaacc
     4681 atataaaaca gctgttatac ccattaatgg ttcacctcga acacccaggc gaggtcagaa
     4741 caggagtgca cggatagcaa aacaactaga aaatgataca agaattattg aagttctctg
     4801 taaagaacat gaatgtaata tagatgaggt gaaaaatgtt tatttcaaaa attttatacc
     4861 ttttatgaat tctcttggac ttgtaacatc taatggactt ccagaggttg aaaatctttc
     4921 taaacgatac gaagaaattt atcttaaaaa taaagatcta gatgcaagat tatttttgga
     4981 tcatgataaa actcttcaga ctgattctat agacagtttt gaaacacaga gaacaccacg
     5041 aaaaagtaac cttgatgaag aggtgaatgt aattcctcca cacactccag ttaggactgt
     5101 tatgaacact atccaacaat taatgatgat tttaaattca gcaagtgatc aaccttcaga
     5161 aaatctgatt tcctatttta acaactgcac agtgaatcca aaagaaagta tactgaaaag
     5221 agtgaaggat ataggataca tctttaaaga gaaatttgct aaagctgtgg gacagggttg
     5281 tgtcgaaatt ggatcacagc gatacaaact tggagttcgc ttgtattacc gagtaatgga
     5341 atccatgctt aaatcagaag aagaacgatt atccattcaa aattttagca aacttctgaa
     5401 tgacaacatt tttcatatgt ctttattggc gtgcgctctt gaggttgtaa tggccacata
     5461 tagcagaagt acatctcaga atcttgattc tggaacagat ttgtctttcc catggattct
     5521 gaatgtgctt aatttaaaag cctttgattt ttacaaagtg atcgaaagtt ttatcaaagc
     5581 agaaggcaac ttgacaagag aaatgataaa acatttagaa cgatgtgaac atcgaatcat
     5641 ggaatccctt gcatggctct cagattcacc tttatttgat cttattaaac aatcaaagga
     5701 ccgagaagga ccaactgatc accttgaatc tgcttgtcct cttaatcttc ctctccagaa
     5761 taatcacact gcagcagata tgtatctttc tcctgtaaga tctccaaaga aaaaaggttc
     5821 aactacgcgt gtaaattcta ctgcaaatgc agagacacaa gcaacctcag ccttccagac
     5881 ccagaagcca ttgaaatcta cctctctttc actgttttat aaaaaagtgt atcggctagc
     5941 ctatctccgg ctaaatacac tttgtgaacg ccttctgtct gagcacccag aattagaaca
     6001 tatcatctgg acccttttcc agcacaccct gcagaatgag tatgaactca tgagagacag
     6061 gcatttggac caaattatga tgtgttccat gtatggcata tgcaaagtga agaatataga
     6121 ccttaaattc aaaatcattg taacagcata caaggatctt cctcatgctg ttcaggagac
     6181 attcaaacgt gttttgatca aagaagagga gtatgattct attatagtat tctataactc
     6241 ggtcttcatg cagagactga aaacaaatat tttgcagtat gcttccacca ggccccctac
     6301 cttgtcacca atacctcaca ttcctcgaag cccttacaag tttcctagtt cacccttacg
     6361 gattcctgga gggaacatct atatttcacc cctgaagagt ccatataaaa tttcagaagg
     6421 tctgccaaca ccaacaaaaa tgactccaag atcaagaatc ttagtatcaa ttggtgaatc
     6481 attcgggact tctgagaagt tccagaaaat aaatcagatg gtatgtaaca gcgaccgtgt
     6541 gctcaaaaga agtgctgaag gaagcaaccc tcctaaacca ctgaaaaaac tacgctttga
     6601 tattgaagga tcagatgaag cagatggaag taaacatctc ccaggagagt ccaaatttca
     6661 gcagaaactg gcagaaatga cttctactcg aacacgaatg caaaagcaga aaatgaatga
     6721 tagcatggat acctcaaaca aggaagagaa atgagaattc cccgatatcg cctcgagtct
     6781 agatccagat cttattaaag cagaacttgt ttattgcagc ttataatggt tacaaataaa
     6841 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     6901 tgtccaaact catcaatgta tcttatcatg tctggtcgac tctagact
//
