LOCUS       sequence-10540-e        5830 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 5830)
  AUTHORS   Zhang GJ, Safran M, Wei W, Sorensen E, Lassota P, Zhelev N, Neuberg 
            DS, Shapiro G, Kaelin WG
  TITLE     Bioluminescent imaging of Cdk2 inhibition in vivo.
  JOURNAL   Nat Med. 2004 Jun . 10(6):643-8.
  PUBMED    15122251
REFERENCE   2  (bases 1 to 5830)
  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: "Nat Med. 
            2004 Jun . 10(6):643-8."
COMMENT     Sequence Label: pGL3-p27Luc
FEATURES             Location/Qualifiers
     source          1..5830
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        48..244
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      95..230
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             854..2506
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(920..938)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2547..2668
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2558..2577)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2612..2631
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2990..3010
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     complement(3162..3179)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3333..3921)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3413..3432)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4092..4883,4884..4952))
                     /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     4715..4734
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4953..5057)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5084..5539
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5171..5190)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5381..5402
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5670..5718
                     /note="synthetic polyadenylation signal"
     misc_feature    5732..5823
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5772..5791
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcccgg gctcgagatc tgcgatctgc atctcaatta
       61 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
      121 cgcccattct ccgccccatc gctgactaat tttttttatt tatgcagagg ccgaggccgc
      181 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
      241 caaaaagctt gccaccatgt caaacgtgcg agtgtctaac gggagcccta gcctggagcg
      301 gatggacgcc aggcaggcgg agcaccccaa gccctcggcc tgcaggaacc tcttcggccc
      361 ggtggaccac gaagagttaa cccgggactt ggagaagcac tgcagagaca tggaagaggc
      421 gagccagcgc aagtggaatt tcgattttca gaatcacaaa cccctagagg gcaagtacga
      481 gtggcaagag gtggagaagg gcagcttgcc cgagttctac tacagacccc cgcggccccc
      541 caaaggtgcc tgcaaggtgc cggcgcagga gagccaggat gtcagcggga gccgcccggc
      601 ggcgccttta attggggctc cggctaactc tgaggacacg catttggtgg acccaaagac
      661 tgatccgtcg gacagccaga cggggttagc ggagcaatgc gcaggaataa ggaagcgacc
      721 tgcaaccgac gattcttcta ctcaaaacaa aagagccaac agaacagaag aaaatgtttc
      781 agacggttcc ccaaatgccg gttctgtgga gcagacgccc aagaagcctg gcctcagaag
      841 acgtcaaacg accatggaag acgccaaaaa cataaagaaa ggcccggcgc cattctatcc
      901 gctggaagat ggaaccgctg gagagcaact gcataaggct atgaagagat acgccctggt
      961 tcctggaaca attgctttta cagatgcaca tatcgaggtg gacatcactt acgctgagta
     1021 cttcgaaatg tccgttcggt tggcagaagc tatgaaacga tatgggctga atacaaatca
     1081 cagaatcgtc gtatgcagtg aaaactctct tcaattcttt atgccggtgt tgggcgcgtt
     1141 atttatcgga gttgcagttg cgcccgcgaa cgacatttat aatgaacgtg aattgctcaa
     1201 cagtatgggc atttcgcagc ctaccgtggt gttcgtttcc aaaaaggggt tgcaaaaaat
     1261 tttgaacgtg caaaaaaagc tcccaatcat ccaaaaaatt attatcatgg attctaaaac
     1321 ggattaccag ggatttcagt cgatgtacac gttcgtcaca tctcatctac ctcccggttt
     1381 taatgaatac gattttgtgc cagagtcctt cgatagggac aagacaattg cactgatcat
     1441 gaactcctct ggatctactg gtctgcctaa aggtgtcgct ctgcctcata gaactgcctg
     1501 cgtgagattc tcgcatgcca gagatcctat ttttggcaat caaatcattc cggatactgc
     1561 gattttaagt gttgttccat tccatcacgg ttttggaatg tttactacac tcggatattt
     1621 gatatgtgga tttcgagtcg tcttaatgta tagatttgaa gaagagctgt ttctgaggag
     1681 ccttcaggat tacaagattc aaagtgcgct gctggtgcca accctattct ccttcttcgc
     1741 caaaagcact ctgattgaca aatacgattt atctaattta cacgaaattg cttctggtgg
     1801 cgctcccctc tctaaggaag tcggggaagc ggttgccaag aggttccatc tgccaggtat
     1861 caggcaagga tatgggctca ctgagactac atcagctatt ctgattacac ccgaggggga
     1921 tgataaaccg ggcgcggtcg gtaaagttgt tccatttttt gaagcgaagg ttgtggatct
     1981 ggataccggg aaaacgctgg gcgttaatca aagaggcgaa ctgtgtgtga gaggtcctat
     2041 gattatgtcc ggttatgtaa acaatccgga agcgaccaac gccttgattg acaaggatgg
     2101 atggctacat tctggagaca tagcttactg ggacgaagac gaacacttct tcatcgttga
     2161 ccgcctgaag tctctgatta agtacaaagg ctatcaggtg gctcccgctg aattggaatc
     2221 catcttgctc caacacccca acatcttcga cgcaggtgtc gcaggtcttc ccgacgatga
     2281 cgccggtgaa cttcccgccg ccgttgttgt tttggagcac ggaaagacga tgacggaaaa
     2341 agagatcgtg gattacgtcg ccagtcaagt aacaaccgcg aaaaagttgc gcggaggagt
     2401 tgtgtttgtg gacgaagtac cgaaaggtct taccggaaaa ctcgacgcaa gaaaaatcag
     2461 agagatcctc ataaaggcca agaagggcgg aaagatcgcc gtgtaattct agagtcgggg
     2521 cggccggccg cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac
     2581 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     2641 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     2701 ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtaa
     2761 aatcgataag gatctgaacg atggagcgga gaatgggcgg aactgggcgg agttaggggc
     2821 gggatgggcg gagttagggg cgggactatg gttgctgact aattgagatg catgctttgc
     2881 atacttctgc ctgctgggga gcctggggac tttccacacc tggttgctga ctaattgaga
     2941 tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca ccctaactga
     3001 cacacattcc acagcggatc cgtcgaccga tgcccttgag agccttcaac ccagtcagct
     3061 ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtc ttctttatca
     3121 tgcaactcgt aggacaggtg ccggcagcgc tcttccgctt cctcgctcac tgactcgctg
     3181 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     3241 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     3301 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     3361 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     3421 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     3481 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     3541 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     3601 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     3661 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     3721 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     3781 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     3841 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     3901 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     3961 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     4021 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     4081 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     4141 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     4201 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     4261 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     4321 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     4381 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     4441 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     4501 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     4561 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     4621 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     4681 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     4741 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     4801 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     4861 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     4921 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     4981 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     5041 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgcgcc ctgtagcggc
     5101 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     5161 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     5221 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     5281 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     5341 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     5401 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     5461 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     5521 atattaacgc ttacaatttg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     5581 tcggtgcggg cctcttcgct attacgccag cccaagctac catgataagt aagtaatatt
     5641 aaggtacggg aggtacttgg agcggccgca ataaaatatc tttattttca ttacatctgt
     5701 gtgttggttt tttgtgtgaa tcgatagtac taacatacgc tctccatcaa aacaaaacga
     5761 aacaaaacaa actagcaaaa taggctgtcc ccagtgcaag tgcaggtgcc agaacatttc
     5821 tctatcgata
//
