LOCUS       sequence-171282-        6178 bp    DNA     circular SYN 29-JUL-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6178)
  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 6178)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 29, 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..6178
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    130..251
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(441..896)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(578..599)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     790..809
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        923..1027
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1028..1096,1097..1888)
                     /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(1246..1265)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2059..2647
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2548..2567
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2801..2818
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    2960..3094
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(3519..3539)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          3713..3778
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(4019..5671)
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     5587..5605
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
ORIGIN
        1 gtctggcgcg cggcggcgag gggcgggccg cgggcgccga gggcggggcc gagcggcgct
       61 ggagcggcaa aaagccgcgt ggctggcgct cggtacctcc cgggttatgt tagctcagtt
      121 acagtaccat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
      181 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
      241 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt
      301 gggaggtttt ttaaagcaag taaaacctcc acgtacctta atattactta cttatcatgg
      361 tagcttgggc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
      421 cagcctgaat ggcgaatggc aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
      481 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
      541 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
      601 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
      661 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
      721 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
      781 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
      841 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg
      901 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
      961 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     1021 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     1081 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     1141 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     1201 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     1261 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     1321 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     1381 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     1441 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     1501 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     1561 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     1621 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     1681 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     1741 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     1801 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     1861 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     1921 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     1981 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     2041 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     2101 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     2161 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     2221 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     2281 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     2341 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     2401 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     2461 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     2521 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     2581 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     2641 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     2701 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     2761 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     2821 agcggaagag cgctgccggc acctgtccta cgagttgcat gataaagaag acagtcataa
     2881 gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc
     2941 tcaagggcat cggtcgacgg atccagacat gataagatac attgatgagt ttggacaaac
     3001 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     3061 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     3121 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     3181 tggtatggct gattatgatc tctagtcaag gcactataca tcaaatattc cttattaacc
     3241 cctttacaaa ttaaaaagct aaaggtacac aatttttgag catagttatt aatagcagac
     3301 actctatgcc tgtgtggagt aagaaaaaac agtatgttat gattataact gttatgccta
     3361 cttataaagg ttacagaata tttttccata attttcttgt atagcagtgc agctttttcc
     3421 tttgtggtgt aaatagcaaa gcaagcaaga gttctattac taaacacagc atgactcaaa
     3481 aaacttagca attctgaagg aaagtccttg gggtcttcta cctttctctt cttttttgga
     3541 ggagtagaat gttgagagtc agcagtagcc tcatcatcac tagatggcat ttcttctgag
     3601 caaaacaggt tttcctcatt aaaggcattc caccactgct cccattcatc agttccatag
     3661 gttggtaaat atctgagaat ccatcttaat aaataaatcc atagtttgga atctaaaata
     3721 cacaaacaat tagaatcagt agtttaacac attatacact taaaaatttt atatttacct
     3781 tagagcttta aatctctgta ggtagtttgt ccaattatgt cacaccacag aagtaaggtt
     3841 ccttcacaaa gatccggcga atttctgcca ttcatccgct tattatcact tattcaggcg
     3901 tagcaaccag gcgtttaagg gcaccaataa ctgccttaaa aaaattacgc cccgccctgc
     3961 cactcatcgc agtattacaa tagctaagaa tttcgtcatc gctgaataca gttacatttt
     4021 acaatttgga ctttccgccc ttcttggcct ttatgaggat ctctctgatt tttcttgcgt
     4081 cgagttttcc ggtaagacct ttcggtactt cgtccacaaa cacaactcct ccgcgcaact
     4141 ttttcgcggt tgttacttga ctggcgacgt aatccacgat ctctttttcc gtcatcgtct
     4201 ttccgtgctc caaaacaaca acggcggcgg gaagttcacc ggcgtcatcg tcgggaagac
     4261 ctgccacgcc cgcgtcgaag atgttggggt gttgtaacaa tatcgattcc aattcagcgg
     4321 gggccacctg atatcctttg tatttaatta aagacttcaa gcggtcaact atgaagaagt
     4381 gttcgtcttc gtcccagtaa gctatgtctc cagaatgtag ccatccatcc ttgtcaatca
     4441 aggcgttggt cgcttccgga ttgtttacat aaccggacat aatcataggt cctctgacac
     4501 ataattcgcc tctctgatta acgcccagcg ttttcccggt atccagatcc acaaccttcg
     4561 cttcaaaaaa tggaacaact ttaccgaccg cgcccggttt atcatccccc tcgggtgtaa
     4621 tcagaatagc tgatgtagtc tcagtgagcc catatccttg tcgtatccct ggaagatgga
     4681 agcgttttgc aaccgcttcc ccgacttctt tcgaaagagg tgcgccccca gaagcaattt
     4741 cgtgtaaatt agataaatcg tatttgtcaa tcagagtgct tttggcgaag aatgaaaata
     4801 gggttggtac tagcaacgca ctttgaattt tgtaatcctg aagggatcgt aaaaacagct
     4861 cttcttcaaa tctatacatt aagacgactc gaaatccaca tatcaaatat ccgagtgtag
     4921 taaacattcc aaaaccgtga tggaatggaa caacacttaa aatcgcagta tccggaatga
     4981 tttgattgcc aaaaatagga tctctggcat gcgagaatct gacgcaggca gttctatgcg
     5041 gaagggccac acccttaggt aacccagtag atccagagga attcattatc agtgcaattg
     5101 ttttgtcacg atcaaaggac tctggtacaa aatcgtattc attaaaaccg ggaggtagat
     5161 gagatgtgac gaacgtgtac atcgactgaa atccctggta atccgtttta gaatccatga
     5221 taataatttt ctggattatt ggtaattttt tttgcacgtt caaaattttt tgcaacccct
     5281 ttttggaaac aaacactacg gtaggctgcg aaatgttcat actgttgagc aattcacgtt
     5341 cattataaat gtcgttcgcg ggcgcaactg caactccgat aaataacgcg cccaacaccg
     5401 gcataaagaa ttgaagagag ttttcactgc atacgacgat tctgtgattt gtattcagcc
     5461 catatcgttt catagcttct gccaaccgaa cggacatttc gaagtattcc gcgtacgtga
     5521 tgttcacctc gatatgtgca tctgtaaaag caattgttcc aggaaccagg gcgtatctct
     5581 tcatagcctt atgcagttgc tctccagcgg ttccatcctc tagaggatag aatggcgccg
     5641 ggcctttctt tatgtttttg gcgtcttcca ttttaccaac agtaccggaa tgccaagctt
     5701 ggggatcctg caaaaactaa catgaagcca cccccatgac cccaaggaaa cacaccaaac
     5761 aagacaaaaa cagagagaag aagagaaagc tgatcaggtt atgagctccg ttcctgccca
     5821 agggaccccc gcccccgtag gcagagccaa gtcgggtacc caggtgagat ggggggggat
     5881 gctccccaac ctgtctcttt tgtactcaca acggtcacgt ttgccttcct cttcctggag
     5941 cgagccgaga actccgcgcc gccgtcctcc ttcatggtgt cccgctcctt cgcatccctg
     6001 tggacccggg tggcaccggg ccgggtgagc acccgcggcc gcccctcccc cacgccaccc
     6061 gtcccgtccc gtcccgtccc gtccggcccc gcggcgcggg cactcaccgc tccctgcgct
     6121 ccctcggcat gatggggctg ctccggcctg aggcctgcgg ggcggcgcgg ggtgaggc
//
