LOCUS       sequence-156184-        4755 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 4755)
  AUTHORS   Mizutani K, Yoon K, Dang L, Tokunaga A, Gaiano N
  TITLE     Differential Notch signalling distinguishes neural stem cells from 
            intermediate progenitors.
  JOURNAL   Nature. 2007 Sep 20. 449(7160):351-5.
  PUBMED    17721509
REFERENCE   2  (bases 1 to 4755)
  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: "Nature. 
            2007 Sep 20. 449(7160):351-5."
COMMENT     Sequence Label: CBFRE-EGFP
FEATURES             Location/Qualifiers
     source          1..4755
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..197
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      48..183
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      219..228
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(225..227,228..230,231..944)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(225..227,228..230,231..941)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(270..291)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(531..550)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     878..899
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1057..1191
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1094..1113)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1148..1167
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    1200..1233
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        complement(1245..1263)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1246..1263)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     repeat_region   1336..1438
                     /label=ITR
                     /note="inverted terminal repeat of human adenovirus 
                     serotype 5"
     rep_origin      1522..1977
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1609..1628)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1819..1840
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2003..2107
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2108..2176,2177..2968)
                     /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(2326..2345)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3139..3727
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3628..3647
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3881..3898
                     /label=L4440
                     /note="L4440 vector, forward primer"
     repeat_region   3977..4079
                     /label=ITR
                     /note="inverted terminal repeat of human adenovirus 
                     serotype 5"
     misc_signal     4167..4317
                     /label=Ad5 Psi
                     /note="packaging signal for adenovirus serotype 5"
     primer_bind     4531..4547
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     4532..4548
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
       61 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
      121 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
      181 gcctaggctt ttgcaaaaag ctgggggatc caccggtcgc caccatggtg agcaagggcg
      241 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
      301 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
      361 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
      421 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
      481 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
      541 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
      601 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
      661 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
      721 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
      781 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
      841 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
      901 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaagcggc cgcgactcta
      961 gaggatcaat tctttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     1021 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     1081 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     1141 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat catcgatcca
     1201 taacttcgta taatgtatgc tatacgaagt tatccagatc tggttctata gtgtcaccta
     1261 aatcgtatgt gtatgataca taaggttatg tattaattgt agccgcgttc taacgacaat
     1321 atgtccatag ggcccctacg tcacccgccc cgttcccacg ccccgcgcca cgtcacaaac
     1381 tccaccccct cattatcata ttggcttcaa tccaaaataa ggtatattat tgatgatggc
     1441 cgcagcggcc ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     1501 gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     1561 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     1621 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     1681 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     1741 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     1801 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     1861 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     1921 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg
     1981 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     2041 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     2101 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     2161 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     2221 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     2281 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     2341 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     2401 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     2461 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     2521 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     2581 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     2641 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     2701 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     2761 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     2821 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     2881 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     2941 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     3001 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     3061 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     3121 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3181 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     3241 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     3301 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3361 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3421 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     3481 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     3541 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     3601 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     3661 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     3721 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     3781 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     3841 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     3901 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     3961 caggggccgc tgcggccatc atcaataata taccttattt tggattgaag ccaatatgat
     4021 aatgaggggg tggagtttgt gacgtggcgc ggggcgtggg aacggggcgg gtgacgtagt
     4081 agtgtggcgg aagtgtgatg ttgcaagtgt ggcggaacac atgtaagcga cggatgtggc
     4141 aaaagtgacg tttttggtgt gcgccggtgt acacaggaag tgacaatttt cgcgcggttt
     4201 taggcggatg ttgtagtaaa tttgggcgta accgagtaag atttggccat tttcgcggga
     4261 aaactgaata agaggaagtg aaatctgaat aattttgtgt tactcatagc gcgtaatatt
     4321 tgtctagggc cgcggggact ttgaccgttt acgtggagac tcgcccaggt gtttttctca
     4381 ggtgttttcc gcgttccggg tcaaagttgg cgttttatta ttatagtcag ctgacgtgta
     4441 gtgtatttat acccggtgag ttcctcaaga ggccactctt gagtgccagc gagtagagtt
     4501 ttctcctccg agccgctccg acaccgggac tcgaggtcga cggtatcgat aagcttgata
     4561 tcgaattcct gcagcccggg aggtaccgag ctcttacgcg tgctagctcg agatctggag
     4621 taacacgccg tgggaaaaaa tttatggatc tggagtaaca cgccgtggga aaaaatttat
     4681 ggatctggag taacacgccg tgggaaaaaa tttatggatc tggagtaaca cgccgtggga
     4741 aaaaatttat ggatc
//
