LOCUS       sequence-333557-        5867 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 5867)
  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 5867)
  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: GLASTp-DsRed2
FEATURES             Location/Qualifiers
     source          1..5867
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..92
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     41..60
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     regulatory      2196..2205
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2202..2879
                     /codon_start=1
                     /product="improved tetrameric variant of DsRed fluorescent 
                     protein"
                     /label=DsRed2
                     /note="mammalian codon-optimized"
                     /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     primer_bind     complement(2382..2402)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     2794..2817
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    2929..3050
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2940..2959)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2994..3013
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3298..3315)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3469..4057)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3549..3568)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4228..5019,5020..5088))
                     /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     4851..4870
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5089..5193)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5220..5675
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5307..5326)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5517..5538
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5806..5854
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 aacatacgct ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc
       61 cagtgcaagt gcaggtgcca gaacatttct ctatcgatag gtaccatgtc tacacaaact
      121 gaaataatga aatggttttt aaccaagctt tagggattag agaggaaaaa aaaagtgaaa
      181 aagaaagggc accagggtct tcccatctct atgctaacca aagtgtggtt ttccagccct
      241 ttttcttgcc tattaacaaa aataaaaaca aaaaatggaa atgccactgg gcttgaaacc
      301 ccatttaaaa tcagatttct ctttcgagtt cccttgcaat acaagtcaaa gtcatttcca
      361 ccaaaaaaaa aaagcggtgg gggggggtgc gcaaattaat cctaaactca tggctcagat
      421 aggacctggt atagtaaata aatgctttta aaagtagtca tggcattgag aaatggctcc
      481 atgcacctaa accagaggca ccaatgtaaa aactagtaat tgtgttttaa cacaaaagat
      541 tgaaaagcgt tttgtttcac atcctaatat tattccaccc acttaaaaag agttacttaa
      601 tattgactct gtattaggta taagacaagt tcttgccctc aaagaactta taatttagtt
      661 gagaaaccaa aacttaacat gagcttaaaa agaaggatgc agtacaaaac atggtataat
      721 taagtgctaa acaatgtagt atagatgctc agtgacctag gagatcagag gaagggaagg
      781 gtcactgagg gctggggatg tcaaaagcat gtactttcag taagttctta aaatgatagc
      841 atgttagaaa gctgttataa ttgctgctga tttgtaattt taaagcgttg acatgattca
      901 ccattgcttt ccaaccagta gctgtttaaa aggacataca gttgagggtg aaaggagata
      961 tttctttatg atctccagag atttgtattg aggggaaaaa agacatactt tcttgttgaa
     1021 taatagacca agaattgagc atattttgca tggggaatat taaattcagt gaacaagaat
     1081 ccaaacctca ggctgtgatt ctggtataaa tttttgtttc taatgaacca ttttcttcct
     1141 tctaaagaag atttaagttt tgggaataag tcacctcaaa aaattttttt tttattttct
     1201 aatatctatt ttcacaattt atttttcttt taagaaaagt ggactagaag ctcaagttac
     1261 cttctcaagg caatgagaat tttttagctc taaaaaagtt gtgtgtggta gggagaacat
     1321 ttagcatcct ttaaaatgga gacatctgaa cagcaaccta tcttttgaat cgccaaatct
     1381 caagtgtact ttgcactctc ggtcttaaaa tgctcttcat tttctagttt ggcaaaaatt
     1441 ccagtctctt tacatttgct accttttccg tctttgcaac acaggcagtg tgagctaaat
     1501 taggtttgtt aaaatgaagt tttgaagttt caactgctga aatagaggca tgtccctaac
     1561 tttagacaaa tttagagaat cgtagtaagc attatttttt ttctacatac ttgtctgcat
     1621 tcagtgtctt cccaagaaac gtcagttaat tccccgattc gaaaaaggaa ctttctcgta
     1681 acagttgtac aaccaggcta aagagcacac cctcgtcttc cctgaaaggg aggaaacatg
     1741 caataatgtg aggtaattaa atttagcctt agaaaagtct ttgaaacgtt ctaatcaaga
     1801 ctttcagctt tccgattgtg tgcgctttat ttttcaattc ttcctagtcc cggttttgca
     1861 tttctaatga cagtgaactg acaaagcgtg aaagtggtct aaggagcaaa acaaagccag
     1921 caagcctgct tctggtgtgc caagattaat caatattcca gaaacctcgg ggtttccccc
     1981 tcctccctgt gcggtttcca ttccccactc ctcctttccc gggcacagac tcctcccctt
     2041 ttcttaagtt gccctgatag taacttgcag tttcagagca catgcacact gtcagggcta
     2101 gcctggctgc ttacgcgcgc tgcggattgt tgctccgttg tacctgctgg ggaattctgc
     2161 agtcgacggt accgcgggcc cgggatccac cggtcgccac catggcctcc tccgagaacg
     2221 tcatcaccga gttcatgcgc ttcaaggtgc gcatggaggg caccgtgaac ggccacgagt
     2281 tcgagatcga gggcgagggc gagggccgcc cctacgaggg ccacaacacc gtgaagctga
     2341 aggtgaccaa gggcggcccc ctgcccttcg cctgggacat cctgtccccc cagttccagt
     2401 acggctccaa ggtgtacgtg aagcaccccg ccgacatccc cgactacaag aagctgtcct
     2461 tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggcgaccg
     2521 tgacccagga ctcctccctg caggacggct gcttcatcta caaggtgaag ttcatcggcg
     2581 tgaacttccc ctccgacggc cccgtgatgc agaagaagac catgggctgg gaggcctcca
     2641 ccgagcgcct gtacccccgc gacggcgtgc tgaagggcga gacccacaag gccctgaagc
     2701 tgaaggacgg cggccactac ctggtggagt tcaagtccat ctacatggcc aagaagcccg
     2761 tgcagctgcc cggctactac tacgtggacg ccaagctgga catcacctcc cacaacgagg
     2821 actacaccat cgtggagcag tacgagcgca ccgagggccg ccaccacctg ttcctgtagc
     2881 ggccgcgact ctagagtcgg ggcggccggc cgcttcgagc agacatgata agatacattg
     2941 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     3001 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     3061 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
     3121 aaaacctcta caaatgtggt aaaatcgata aggatccgtc gaccgatgcc cttgagagcc
     3181 ttcaacccag tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg
     3241 actgtcttct ttatcatgca actcgtagga caggtgccgg cagcgctctt ccgcttcctc
     3301 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     3361 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     3421 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     3481 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     3541 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     3601 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     3661 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     3721 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     3781 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     3841 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     3901 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     3961 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     4021 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     4081 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     4141 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     4201 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     4261 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     4321 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     4381 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     4441 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     4501 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     4561 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     4621 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     4681 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     4741 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     4801 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     4861 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     4921 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     4981 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     5041 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     5101 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     5161 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     5221 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     5281 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     5341 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     5401 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     5461 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     5521 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     5581 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     5641 cgcgaatttt aacaaaatat taacgcttac aatttgccat tcgccattca ggctgcgcaa
     5701 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagccca agctaccatg
     5761 ataagtaagt aatattaagg tacgggaggt acttggagcg gccgcaataa aatatcttta
     5821 ttttcattac atctgtgtgt tggttttttg tgtgaatcga tagtact
//
