LOCUS       sequence-176423-        5784 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 5784)
  AUTHORS   Tran H, Brunet A, Grenier JM, Datta SR, Fornace AJ Jr, DiStefano PS,
            Chiang LW, Greenberg ME
  TITLE     DNA repair pathway stimulated by the forkhead transcription factor 
            FOXO3a through the Gadd45 protein.
  JOURNAL   Science 2002 Apr 19;296(5567):530-4.
  PUBMED    11964479
REFERENCE   2  (bases 1 to 5784)
  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: "Science"; 
            date: "2002-04-19"; volume: "296(5567)"; pages: "530-4"
COMMENT     Sequence Label: GADD45 WT-luc
FEATURES             Location/Qualifiers
     source          1..5784
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    82..130
                     /note="synthetic polyadenylation signal"
     misc_feature    144..235
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     184..203
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     regulatory      1290..1299
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1296..2948
                     /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(1362..1380)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2989..3110
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3000..3019)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3054..3073
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3358..3375)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3529..4117)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3609..3628)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4288..5079,5080..5148))
                     /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     4911..4930
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5149..5253)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5280..5735
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5367..5386)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5577..5598
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ggcctcttcg ctattacgcc agcccaagct accatgataa gtaagtaata ttaaggtacg
       61 ggaggtactt ggagcggccg caataaaata tctttatttt cattacatct gtgtgttggt
      121 tttttgtgtg aatcgatagt actaacatac gctctccatc aaaacaaaac gaaacaaaac
      181 aaactagcaa aataggctgt ccccagtgca agtgcaggtg ccagaacatt tctctatcga
      241 taggtaccga gctcttacgc gtgctagccc gggctcgaga tctgcgatct aagtaagctt
      301 agggcatatc gagagcattt tgaatttaga cttgtgggct gtgtgggtgt cagatggttg
      361 tctctcagct ggtgggcgtc cagaaggatc cttgtttggg caaggctctt tgagaaagga
      421 gaatctgggt tgccagggat tcccacatgt ggtcaccagc tccccacgca gaccagctca
      481 cgatttccca gttacaccgg gcaggtggga aaccgttctg ctttctgtgg aaaagattct
      541 aacttggttc cctgccatcc ctgaatacaa acgggttggt ttttcttttt tgagcttcca
      601 acccttgcag ctttccgaaa ataaatcaaa ccagccatca aggcaccgaa ataatactac
      661 tgctaataag cagcttcgcc tagacttaga taaacaacac ttctgaggta aactttgccc
      721 cggaggtctg gagacacttt tttaatgtaa cctgcttact aataattact agacttcagt
      781 gcattaaccc tggaaataga ttttaatagc caccccttaa aacaaaagac atgaaaagat
      841 aataagaaaa aagtgccgca actattatag aaaaacactt ggcagcctgc ttcagcccaa
      901 gctgaggcca cctctagcct ctgctaaagc cccccactcc caatggtccc cgccaaccgg
      961 ataagagtgc gcgcgggacc cgccttcccc tctcggcacc gcccccgccc ccgccccctc
     1021 ggctcgcctc ccgcgtggct cctccctttt ccgctcctct caacctgact ccaggagctg
     1081 gggtcaaatt gctggagcag gctgatttgc atagcccaat ggccaagctg catgcaaatg
     1141 aggcggaagg tggttggctg agggttggca ggataacccc ggagagcggg gccctttgtc
     1201 ctccagtggc tggtaggcag tggctgggag gagatcaagc ttatgatctg cgatctaagt
     1261 aagcttggca ttccggtact gttggtaaag ccaccatgga agacgccaaa aacataaaga
     1321 aaggcccggc gccattctat ccgctggaag atggaaccgc tggagagcaa ctgcataagg
     1381 ctatgaagag atacgccctg gttcctggaa caattgcttt tacagatgca catatcgagg
     1441 tggacatcac ttacgctgag tacttcgaaa tgtccgttcg gttggcagaa gctatgaaac
     1501 gatatgggct gaatacaaat cacagaatcg tcgtatgcag tgaaaactct cttcaattct
     1561 ttatgccggt gttgggcgcg ttatttatcg gagttgcagt tgcgcccgcg aacgacattt
     1621 ataatgaacg tgaattgctc aacagtatgg gcatttcgca gcctaccgtg gtgttcgttt
     1681 ccaaaaaggg gttgcaaaaa attttgaacg tgcaaaaaaa gctcccaatc atccaaaaaa
     1741 ttattatcat ggattctaaa acggattacc agggatttca gtcgatgtac acgttcgtca
     1801 catctcatct acctcccggt tttaatgaat acgattttgt gccagagtcc ttcgataggg
     1861 acaagacaat tgcactgatc atgaactcct ctggatctac tggtctgcct aaaggtgtcg
     1921 ctctgcctca tagaactgcc tgcgtgagat tctcgcatgc cagagatcct atttttggca
     1981 atcaaatcat tccggatact gcgattttaa gtgttgttcc attccatcac ggttttggaa
     2041 tgtttactac actcggatat ttgatatgtg gatttcgagt cgtcttaatg tatagatttg
     2101 aagaagagct gtttctgagg agccttcagg attacaagat tcaaagtgcg ctgctggtgc
     2161 caaccctatt ctccttcttc gccaaaagca ctctgattga caaatacgat ttatctaatt
     2221 tacacgaaat tgcttctggt ggcgctcccc tctctaagga agtcggggaa gcggttgcca
     2281 agaggttcca tctgccaggt atcaggcaag gatatgggct cactgagact acatcagcta
     2341 ttctgattac acccgagggg gatgataaac cgggcgcggt cggtaaagtt gttccatttt
     2401 ttgaagcgaa ggttgtggat ctggataccg ggaaaacgct gggcgttaat caaagaggcg
     2461 aactgtgtgt gagaggtcct atgattatgt ccggttatgt aaacaatccg gaagcgacca
     2521 acgccttgat tgacaaggat ggatggctac attctggaga catagcttac tgggacgaag
     2581 acgaacactt cttcatcgtt gaccgcctga agtctctgat taagtacaaa ggctatcagg
     2641 tggctcccgc tgaattggaa tccatcttgc tccaacaccc caacatcttc gacgcaggtg
     2701 tcgcaggtct tcccgacgat gacgccggtg aacttcccgc cgccgttgtt gttttggagc
     2761 acggaaagac gatgacggaa aaagagatcg tggattacgt cgccagtcaa gtaacaaccg
     2821 cgaaaaagtt gcgcggagga gttgtgtttg tggacgaagt accgaaaggt cttaccggaa
     2881 aactcgacgc aagaaaaatc agagagatcc tcataaaggc caagaagggc ggaaagatcg
     2941 ccgtgtaatt ctagagtcgg ggcggccggc cgcttcgagc agacatgata agatacattg
     3001 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     3061 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     3121 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
     3181 aaaacctcta caaatgtggt aaaatcgata aggatccgtc gaccgatgcc cttgagagcc
     3241 ttcaacccag tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg
     3301 actgtcttct ttatcatgca actcgtagga caggtgccgg cagcgctctt ccgcttcctc
     3361 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     3421 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     3481 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     3541 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     3601 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     3661 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     3721 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     3781 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     3841 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     3901 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     3961 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     4021 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     4081 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     4141 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     4201 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     4261 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     4321 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     4381 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     4441 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     4501 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     4561 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     4621 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     4681 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     4741 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     4801 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     4861 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     4921 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     4981 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     5041 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     5101 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     5161 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     5221 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     5281 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     5341 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     5401 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     5461 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     5521 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     5581 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     5641 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     5701 cgcgaatttt aacaaaatat taacgcttac aatttgccat tcgccattca ggctgcgcaa
     5761 ctgttgggaa gggcgatcgg tgcg
//
