LOCUS       sequence-420598-        7006 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 7006)
  AUTHORS   Geng Y, Eaton EN, Picon M, Roberts JM, Lundberg AS, Gifford A, 
            Sardet C, Weinberg RA
  TITLE     Regulation of cyclin E transcription by E2Fs and retinoblastoma 
            protein.
  JOURNAL   Oncogene 1996 Mar 21;12(6):1173-80.
  PUBMED    8649818
REFERENCE   2  (bases 1 to 7006)
  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: "Oncogene"; 
            date: "1996-03-21"; volume: "12(6)"; pages: "1173-80"
COMMENT     Sequence Label: 10-4 cyclin E promoter
FEATURES             Location/Qualifiers
     source          1..7006
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1429..3081
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(1495..1513)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     intron          3322..3387
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             3561..3581
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    4006..4140
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4282..4299)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4453..5041)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4533..4552)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5212..6003,6004..6072))
                     /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     5835..5854
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6073..6177)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6204..6659
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6291..6310)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6501..6522
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    6849..6970
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 ccgagcgcca gccacgcggc tttttgccgc tccagcgccg ctcggccccg ccctcggcgc
       61 ccgcggcccg cccctcgccg ccgcgcgcca gacttctccc gcgtcccgcc cgccgccccg
      121 ccccgcgtcc cgcgccccgc gccccgcgcc cggccctcgg cgcgcaggcc ctgtcacttg
      181 gccccgccct gtccgccggc cccgcccctg attccccgtc cctgcgcctc gctggccggc
      241 gccgtggagg ggcgggtccg ggggcggggc gagggacggg gcgggacggg ctctgggtcc
      301 cgcgcggccg ctgaggggct gggagccgcg gcggggcggt gcgagggcgg gccggggccg
      361 gttccgcgcg cagggatttt aaatgtcccg ctctgagccg ggcgcaggag cagccggcgc
      421 ggccgccagc gcggtgtagg gggcaggcgc ggatcccgcc accgccgcgc gctcggcccg
      481 ccgactcccg gcgccgccgc cgccactgcc gtcgccgccg ccgcctgccg ggactggagc
      541 gcgccgtccg ccgcggacaa gaccctggtg agtctactgc cgcgggctgc ggacagggac
      601 tcctggggcc cagaccgcgc cggggagctg ggtgggggcg gggtgcggcc gggtcgcggg
      661 tccccgggcg gcgtcgcggg gcgcccggga gggtgctgag agcgcggcgg ggtggggtgg
      721 aggggacact gaggcagccc gggccccggg aggcgacggg ccggtgaggg cggcgggggc
      781 ggcgggaccc cgggtcgggg gtcggcgggt gcccgggtcg cgggtgacag gccaccccgc
      841 catcggccat cttcctggcc cgcccggccg cccgcgcgca aagggggagg ggtactgggc
      901 ccgcggcctg accccgccgc cgcctcaccc cgcgccgccc cgcaggcctc aggccggagc
      961 agccccatca tgccgaggga gcgcagggag cggtgagtgc ccgcgccgcg gggccggacg
     1021 ggacgggacg ggacgggacg ggtggcgtgg gggaggggcg gccgcgggtg ctcacccggc
     1081 ccggtgccac ccgggtccac agggatgcga aggagcggga caccatgaag gaggacggcg
     1141 gcgcggagtt ctcggctcgc tccaggaaga ggaaggcaaa cgtgaccgtt gtgagtacaa
     1201 aagagacagg ttggggagca tcccccccca tctcacctgg gtacccgact tggctctgcc
     1261 tacgggggcg ggggtccctt gggcaggaac ggagctcata acctgatcag ctttctcttc
     1321 ttctctctgt ttttgtcttg tttggtgtgt ttccttgggg tcatgggggt ggcttcatgt
     1381 tagtttttgc aggatcccca agcttggcat tccggtactg ttggtaaaat ggaagacgcc
     1441 aaaaacataa agaaaggccc ggcgccattc tatcctctag aggatggaac cgctggagag
     1501 caactgcata aggctatgaa gagatacgcc ctggttcctg gaacaattgc ttttacagat
     1561 gcacatatcg aggtgaacat cacgtacgcg gaatacttcg aaatgtccgt tcggttggca
     1621 gaagctatga aacgatatgg gctgaataca aatcacagaa tcgtcgtatg cagtgaaaac
     1681 tctcttcaat tctttatgcc ggtgttgggc gcgttattta tcggagttgc agttgcgccc
     1741 gcgaacgaca tttataatga acgtgaattg ctcaacagta tgaacatttc gcagcctacc
     1801 gtagtgtttg tttccaaaaa ggggttgcaa aaaattttga acgtgcaaaa aaaattacca
     1861 ataatccaga aaattattat catggattct aaaacggatt accagggatt tcagtcgatg
     1921 tacacgttcg tcacatctca tctacctccc ggttttaatg aatacgattt tgtaccagag
     1981 tcctttgatc gtgacaaaac aattgcactg ataatgaatt cctctggatc tactgggtta
     2041 cctaagggtg tggcccttcc gcatagaact gcctgcgtca gattctcgca tgccagagat
     2101 cctatttttg gcaatcaaat cattccggat actgcgattt taagtgttgt tccattccat
     2161 cacggttttg gaatgtttac tacactcgga tatttgatat gtggatttcg agtcgtctta
     2221 atgtatagat ttgaagaaga gctgttttta cgatcccttc aggattacaa aattcaaagt
     2281 gcgttgctag taccaaccct attttcattc ttcgccaaaa gcactctgat tgacaaatac
     2341 gatttatcta atttacacga aattgcttct gggggcgcac ctctttcgaa agaagtcggg
     2401 gaagcggttg caaaacgctt ccatcttcca gggatacgac aaggatatgg gctcactgag
     2461 actacatcag ctattctgat tacacccgag ggggatgata aaccgggcgc ggtcggtaaa
     2521 gttgttccat tttttgaagc gaaggttgtg gatctggata ccgggaaaac gctgggcgtt
     2581 aatcagagag gcgaattatg tgtcagagga cctatgatta tgtccggtta tgtaaacaat
     2641 ccggaagcga ccaacgcctt gattgacaag gatggatggc tacattctgg agacatagct
     2701 tactgggacg aagacgaaca cttcttcata gttgaccgct tgaagtcttt aattaaatac
     2761 aaaggatatc aggtggcccc cgctgaattg gaatcgatat tgttacaaca ccccaacatc
     2821 ttcgacgcgg gcgtggcagg tcttcccgac gatgacgccg gtgaacttcc cgccgccgtt
     2881 gttgttttgg agcacggaaa gacgatgacg gaaaaagaga tcgtggatta cgtcgccagt
     2941 caagtaacaa ccgcgaaaaa gttgcgcgga ggagttgtgt ttgtggacga agtaccgaaa
     3001 ggtcttaccg gaaaactcga cgcaagaaaa atcagagaga tcctcataaa ggccaagaag
     3061 ggcggaaagt ccaaattgta aaatgtaact gtattcagcg atgacgaaat tcttagctat
     3121 tgtaatactg cgatgagtgg cagggcgggg cgtaattttt ttaaggcagt tattggtgcc
     3181 cttaaacgcc tggttgctac gcctgaataa gtgataataa gcggatgaat ggcagaaatt
     3241 cgccggatct ttgtgaagga accttacttc tgtggtgtga cataattgga caaactacct
     3301 acagagattt aaagctctaa ggtaaatata aaatttttaa gtgtataatg tgttaaacta
     3361 ctgattctaa ttgtttgtgt attttagatt ccaaactatg gatttattta ttaagatgga
     3421 ttctcagata tttaccaacc tatggaactg atgaatggga gcagtggtgg aatgccttta
     3481 atgaggaaaa cctgttttgc tcagaagaaa tgccatctag tgatgatgag gctactgctg
     3541 actctcaaca ttctactcct ccaaaaaaga agagaaaggt agaagacccc aaggactttc
     3601 cttcagaatt gctaagtttt ttgagtcatg ctgtgtttag taatagaact cttgcttgct
     3661 ttgctattta caccacaaag gaaaaagctg cactgctata caagaaaatt atggaaaaat
     3721 attctgtaac ctttataagt aggcataaca gttataatca taacatactg ttttttctta
     3781 ctccacacag gcatagagtg tctgctatta ataactatgc tcaaaaattg tgtaccttta
     3841 gctttttaat ttgtaaaggg gttaataagg aatatttgat gtatagtgcc ttgactagag
     3901 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     3961 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     4021 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     4081 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc
     4141 cgtcgaccga tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc
     4201 atgactatcg tcgccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg
     4261 ccggcagcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     4321 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     4381 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4441 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4501 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4561 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4621 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4681 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4741 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4801 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4861 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     4921 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4981 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5041 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5101 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     5161 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     5221 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     5281 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     5341 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     5401 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     5461 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     5521 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     5581 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     5641 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     5701 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     5761 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     5821 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     5881 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     5941 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6001 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6061 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     6121 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     6181 atttccccga aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     6241 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     6301 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     6361 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     6421 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     6481 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     6541 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     6601 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaatttg
     6661 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     6721 attacgccag cccaagctac catgataagt aagtaatatt aaggtacgtg gaggttttac
     6781 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     6841 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     6901 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     6961 atgtatctta tggtactgta actgagctaa cataacccgg gaggta
//
