LOCUS       sequence-322126-        7218 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dominant-negative construct to inhibit Notch signaling.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7218)
  AUTHORS   Chen X, Emerson MM
  TITLE     Notch signaling represses cone photoreceptor formation through the 
            regulation of retinal progenitor cell states.
  JOURNAL   Sci Rep. 2021 Jul 15;11(1):14525. doi: 10.1038/s41598-021-93692-w.
  PUBMED    34267251
REFERENCE   2  (bases 1 to 7218)
  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: "Sci Rep."; 
            date: "2021-07-15"; volume: "11(1):14525. doi"; pages: " 
            10.1038/s41598-021-93692-w"
COMMENT     Sequence Label: CAG-EnRRbpj
FEATURES             Location/Qualifiers
     source          1..7218
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..657
                     /label=chicken beta-actin promoter
     intron          658..1674
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1598..1620
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1682..1701
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             2656..2685
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(4236..4255)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4301..4356
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4355..4374)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4717..4733)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4717..4733)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4730..4752)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4741..4757
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4765..4771,4772..4789,4790..4795))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4810..4831
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4890..5085
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4936..5071
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5091..5225
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5128..5147)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5182..5201
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5293..5310)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5464..6052)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5544..5563)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6223..7014,7015..7083))
                     /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     6846..6865
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7084..7188)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
      421 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
      481 tgggggcggg gggggggggg gcgcgcgccg gggggggggg gggggggggg gggggggggc
      541 ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct
      601 tttatggcga ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga
      661 gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc
      721 cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg
      781 ggctgtaatt agcgcttggt ttaatgacgg cttgtttctt ttctgtggct gcgtgaaagc
      841 cttgaggggc tccgggaggg ccctttgtgc ggggggagcg gctcgggggg tgcgtgcgtg
      901 tgtgtgtgcg tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc
      961 gggcgcggcg cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc
     1081 gtgggggggt gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccgggggggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc
     1321 tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag
     1381 ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc
     1441 tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc
     1501 cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg
     1561 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
     1621 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
     1681 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattcac catggtggaa
     1741 tttcgcgatg ccctggagga tcgctgcagc ccacagtcag cgcccagccc cattacccta
     1801 caaatgcagc atcttcacca ccagcaacag cagcagcagc aacagcagca gcaaatgcag
     1861 cacctccacc agctgcagca actgcagcag ttgcaccaac agcaactggc cgccggtgtc
     1921 ttccaccatc cggcaatggc cttcgatgcc gctgcagccg ccgctgctgc agctgctgct
     1981 gcggccgccc acgctcatgc tgctgcactg cagcagcgcc tcagtggcag tggatcgccc
     2041 gcatcctgct ccacgcccgc ctcgtccacg ccgctgacca tcaaggagga ggaaagcgac
     2101 tccgtgatcg gtgacatgag tttccacaat cagacgcaca ccaccaacga ggaggaggag
     2161 gcggaggagg atgacgacat tgatgtggat gtggatgata cgtcggcggg cggacgcctg
     2221 ccaccacccg cccaccagca gcagtcgacg gccaagccct cgctggcctt ttccatctcc
     2281 aacatcctga gcgatcgttt cggagatgtc cagaagcctg gcaagtcgat tgagaaccag
     2341 gccagcatat tccgcccctt cgaggcgaat cgctcccaga ctgccacgcc ctccgccttt
     2401 acaagagtgg atctgctgga gtttagccgg caacagcaag ctgccgccgc agccgctact
     2461 gcggccatga tgctggaacg ggccaacttc cttaactgct tcaatccggc tgcctatccc
     2521 aggatacacg aggaaatcgt gcagagtcgc ctgcgcagga gtgcagccaa tgccgtcatc
     2581 ccgccgccca tgagctccaa gatgagcgat gccaatccag agaaatctgc tctgggatct
     2641 atgcagccca agcttgagca gaagctgatc tccgaggagg acctggcgcc tgttgtgaca
     2701 gggaagtttg gtgagcggcc tccacccaaa cgactcacta gggaagctat gcgaaattat
     2761 ttaaaagaac gaggggatca aacagtgctc attcttcatg caaaagttgc acagaagtct
     2821 tacggaaatg aaaaacgatt tttttgccct cctccttgtg tgtatcttat gggcagtggt
     2881 tggaagaaaa aaaaagagca aatggaacga gatggatgtt ctgaacaaga gtctcaaccc
     2941 tgtgcgttta ttggaatagg aaacagtgac caagaaatgc agcagctgaa cttggaaggg
     3001 aagaactact gcacagccaa aacattgtac atatctgatt cagacaagag aaaacatttc
     3061 atgttgtctg taaagatgtt ctatggcaac agcgatgaca ttggtgtgtt cctcagcaag
     3121 cggataaagg tcatctccaa accctccaaa aagaagcagt cactgaagaa tgctgacttg
     3181 tgcattgctt caggaacgaa ggtggcactg ttcaatcgcc ttcggtccca gacagttagt
     3241 accaggtacc tgcatgtaga aggagggaat ttccacgcca gttcacaaca gtggggagca
     3301 ttttacatcc atctcttgga cgacgacgag tcggaaggag aggagttcac agttagagat
     3361 ggctacatcc attacgggca gactgtcaag cttgtgtgct cagtgactgg catggcactc
     3421 ccaagattga taattaggaa agttgataag cagacggcat tactggatgc agacgaccct
     3481 gtatcacaac tccacaaatg tgcattttac cttaaggata cagaaagaat gtacttgtgc
     3541 ctttctcaag aaagaataat ccaatttcag gccactccat gtccaaaaga acaaaataag
     3601 gaaatgataa acgatggagc ttcctggaca atcattagca cagacaaggc agaatacacg
     3661 ttctatgagg gaatgggccc cgtccttgcc ccagtcaccc ctgtgcctgt cgtagaaagt
     3721 cttcagttga atggcggcgg ggacgtagca atgcttgaac ttacaggaca aaactttact
     3781 ccaaatttac gagtgtggtt tggggatgta gaagccgaaa caatgtacag atgtggagag
     3841 agcatgctct gtgtggtccc agacatttct gcattccggg aaggttggag atgggtccgc
     3901 cagccagtcc aggttccagt gactttggtc cgaaatgacg gggtcattta ctccaccagc
     3961 cttaccttca cctacacacc agagccaggg ccgaggccac actgcagcgc tgcaggagcg
     4021 attctcagag ccaactccag ccaagtgccc tccaatgagt caaacacaaa cagcgagggg
     4081 aattacacaa atgccagcac aaattctacc agtgtcacat cgtccacagc aaccgtggtg
     4141 tcctaagaat tcggccgcac tcctcaggtg caggctgcct atcagaaggt ggtggctggt
     4201 gtggccaatg ccctggctca caaataccac tgagatcttt ttccctctgc caaaaattat
     4261 ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc
     4321 attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca
     4381 aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata tgcccatatg
     4441 ctggctgcca tgaacaaagg ttggctataa agaggtcatc agtatatgaa acagccccct
     4501 gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt ttttatattt
     4561 tgttttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt tttactagcc
     4621 agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct cttatggaga
     4681 tccctcgacc tgcagcccaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     4741 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     4801 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     4861 gtcgggaaac ctgtcgtgcc agcggatccg catctcaatt agtcagcaac catagtcccg
     4921 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4981 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     5041 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct aacttgttta
     5101 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     5161 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     5221 ggatccgctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     5281 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     5341 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5401 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5461 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5521 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5581 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5641 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5701 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     5761 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5821 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     5881 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     5941 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6001 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6061 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6121 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6181 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     6241 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6301 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6361 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     6421 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     6481 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     6541 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     6601 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6661 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     6721 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     6781 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6841 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6901 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6961 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     7021 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     7081 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7141 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     7201 aaaagtgcca cctgggtc
//
