LOCUS       sequence-456543-        6045 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 6045)
  AUTHORS   Kopan R, Schroeter EH, Weintraub H, Nye JS
  TITLE     Signal transduction by activated mNotch: importance of proteolytic 
            processing and its regulation by the extracellular domain.
  JOURNAL   Proc Natl Acad Sci U S A. 1996 Feb 20;93(4):1683-8.
  PUBMED    8643690
REFERENCE   2  (bases 1 to 6045)
  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: "Proc Natl 
            Acad Sci U S A."; date: "1996-02-20"; volume: "93(4)"; pages: 
            "1683-8"
COMMENT     Sequence Label: pCS2 Notch1 Delta-EMV-6MT
FEATURES             Location/Qualifiers
     source          1..6045
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     1759..1775
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1760..1776
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1790..1819
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1829..1858
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1868..1897
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1907..1936
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1946..1975
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2009..2038
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2057..2073)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2057..2073)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     polyA_signal    2097..2231
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2121..2140)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2175..2194
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2344..2364)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2344..2362)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2383..2399)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2383..2399)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2396..2418)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2407..2423
                     /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(2431..2437,2438..2455,2456..2461))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2476..2497
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2614..2631)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2785..3373)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2865..2884)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3544..4335,4336..4404))
                     /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     4167..4186
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4405..4509)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4535..4990)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4672..4693)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4884..4903
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5061..6045
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atccactagt aacggccgcc agtgtgctgg aattccgggt
      121 gggcggcgct gggctccggt ggtgtgcgtc aacgtccgat ccccgccggc caccccaaga
      181 ggccgccgcc cgggctgcgg gcagctggcg agcaggcatg ccacggctcc tgacgccctt
      241 gctctgccta acgctgctgc ccgcgctcgc cgcaagaggc ttgagaatcg attcgaattc
      301 aaggattcct tacaagattg aggccgtgaa gagtgagccg gtggagcctc cgctgccctc
      361 gcagctgcac ctcgtgtacg tggcagcggc cgccttcgtg ctcctgttct ttgtgggctg
      421 tggggtgctg ctgtcccgca agcgccggcg gcagcatggc cagctctggt tccctgaggg
      481 tttcaaagtg tcagaggcca gcaagaagaa gcggagagag cccctcggcg aggactcagt
      541 cggcctcaag cccctgaaga atgcctcaga tggtgctctg atggacgaca atcagaacga
      601 gtggggagac gaagacctgg agaccaagaa gttccggttt gaggagccag tagttctccc
      661 tgacctgagt gatcagactg accacagaca gtggacccag cagcacctgg acgctgctga
      721 cctgcgcatg tctgccatgg ccccaacacc gcctcagggg gaggtggatg ctgactgcat
      781 ggatgtcaat gttcgaggac cagatggctt cacacccctc atgattgcct cctgcagtgg
      841 agggggcctt gagacaggca acagtgaaga agaagaagat gcacctgctg tcatctctga
      901 cttcatctac cagggcgcca gcttgcacaa ccagacagac cgcaccgggg agaccgcctt
      961 gcacttggct gcccgatact ctcgttcaga tgctgcaaag cgcttgctgg aggccagtgc
     1021 agatgccaac atccaggaca acatgggccg tactccgtta catgcagcag tttctgcaga
     1081 tgctcagggt gtcttccaga tcctgctccg gaacagggcc acagatctgg atgcccgaat
     1141 gcatgatggc acaactccac tgatcctggc tgcgcgcctg gccgtggagg gcatgctgga
     1201 ggacctcatc aactcacatg ctgacgtcaa tgccgtggat gacctaggca agtcggcttt
     1261 gcattgggcg gccgcggtga acaatgtgga tgctgctgtt gtgctcctga agaacggagc
     1321 caacaaggac atgcagaaca acaaggagga gactcccctg ttcctggccg cccgtgaggg
     1381 cagctatgag actgccaaag tgttgctgga ccactttgcc aaccgggaca tcacggatca
     1441 catggaccga ttgccgcggg acatcgcaca ggagcgtatg caccacgata tcgtgcggct
     1501 tttggatgag tacaacctgg tgcgcagccc acagctgcat ggcactgccc tgggtggcac
     1561 acccactctg tctcccacac tctgctcgcc caatggctac ctgggcaatc tcaagtctgc
     1621 cacacagggc aagaaggccc gcaagcccag caccaaaggg ctggcttgtg gtagcaagga
     1681 agctaaggac ctcaaggcac ggaggaagaa gtcccaggat ggcaagggct gcctgttgga
     1741 cagctcgatg acgcgtcatc gaggtcgacg gtatcgattt aaagctatgg agcaaaagct
     1801 catttctgaa gaggacttga atgaaatgga gcaaaagctc atttctgaag aggacttgaa
     1861 tgaaatggag caaaagctca tttctgaaga ggacttgaat gaaatggagc aaaagctcat
     1921 ttctgaagag gacttgaatg aaatggagca aaagctcatt tctgaagagg acttgaatga
     1981 aatggagagc ttgggcgacc tcaccatgga gcaaaagctc atttctgaag aggacttgaa
     2041 ttcctgcagc ccgggggatc cactagttct agaactatag tgagtcgtat tacgtagatc
     2101 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     2161 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     2221 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt
     2281 gggaggtttt ttaattcgcg gccgcggcgc caatgcattg ggcccggtac ccagcttttg
     2341 ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt
     2401 gtgaaattgt tatccgctca caattccaca caacatacga gccgggagca taaagtgtaa
     2461 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     2521 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     2581 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     2641 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     2701 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     2761 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     2821 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     2881 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     2941 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     3001 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     3061 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     3121 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     3181 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     3241 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     3301 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     3361 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     3421 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     3481 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     3541 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     3601 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     3661 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     3721 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     3781 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     3841 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     3901 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     3961 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     4021 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     4081 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     4141 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     4201 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     4261 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     4321 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     4381 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     4441 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     4501 ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta
     4561 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     4621 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     4681 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     4741 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     4801 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     4861 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     4921 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     4981 cagggcgcgt cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     5041 gcctcttcgc tattacgcca gtcgaccgcc aattcaatat ggcgtatatg gactcatgcc
     5101 aattcaatat ggtggatctg gacctgtgcc aattcaatat ggcgtatatg gactcgtgcc
     5161 aattcaatat ggtggatctg gaccccagcc aattcaatat ggcggacttg gcaccatgcc
     5221 aattcaatat ggcggacctg gcactgtgcc aactggggag gggtctactt ggcacggtgc
     5281 caagtttgag gaggggtctt ggccctgtgc caagtccgcc atattgaatt ggcatggtgc
     5341 caataatggc ggccatattg gctatatgcc aggatcaata tataggcaat atccaatatg
     5401 gccctatgcc aatatggcta ttggccaggt tcaatactat gtattggccc tatgccatat
     5461 agtattccat atatgggttt tcctattgac gtagatagcc cctcccaatg ggcggtccca
     5521 tataccatat atggggcttc ctaataccgc ccatagccac tcccccattg acgtcaatgg
     5581 tctctatata tggtctttcc tattgacgtc atatgggcgg tcctattgac gtatatggcg
     5641 cctcccccat tgacgtcaat tacggtaaat ggcccgcctg gctcaatgcc cattgacgtc
     5701 aataggacca cccaccattg acgtcaatgg gatggctcat tgcccattca tatccgttct
     5761 cacgccccct attgacgtca atgacggtaa atggcccact tggcagtaca tcaatatcta
     5821 ttaatagtaa cttggcaagt acattactat tggaagtacg ccagggtaca ttggcagtac
     5881 tcccattgac gtcaatggcg gtaaatggcc cgcgatggct gccaagtaca tccccattga
     5941 cgtcaatggg gaggggcaat gacgcaaatg ggcgttccat tgacgtaaat gggcggtagg
     6001 cgtgcctaat gggaggtcta tataagcaat gctcgtttag ggaac
//
