LOCUS       sequence-412653-        6658 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 6658)
  AUTHORS   Ong CT, Cheng HT, Chang LW, Ohtsuka T, Kageyama R, Stormo GD, Kopan 
            R
  TITLE     Target selectivity of vertebrate notch proteins. Collaboration 
            between discrete domains and CSL-binding site architecture 
            determines activation probability.
  JOURNAL   J Biol Chem. 2006 Feb 24. 281(8):5106-19.
  PUBMED    16365048
REFERENCE   2  (bases 1 to 6658)
  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: "J Biol 
            Chem. 2006 Feb 24. 281(8):5106-19."
COMMENT     Sequence Label: 3XFlagNICD3
FEATURES             Location/Qualifiers
     source          1..6658
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             614..679
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    2682..3304
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2881..2900)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3328..3348)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3333..3662
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3513..3648
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3700..3719)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3701..3719)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3733..3749)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3733..3749)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3746..3768)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3757..3773
                     /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(3781..3787,3788..3805,3806..3811))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3826..3847
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3964..3981)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4135..4723)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4215..4234)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4894..5685,5686..5754))
                     /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     5517..5536
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5755..5859)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5886..6341
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5973..5992)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6183..6204
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6467..6489
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6481..6498
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6482..6498
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
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 ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agaattcacc atggactaca aagaccatga cggtgattat aaagatcatg atatcgatta
      661 caaggatgac gatgacaagc ttgcatgcct gcaggtcgac tctagaatgg ttgccaggcg
      721 aaagcgagaa cacagcaccc tctggttccc tgagggtttt gcattacaca aggacatagc
      781 tgctggccac aagggccgga gggagcctgt gggacaagat gcactgggaa tgaagaacat
      841 ggccaagggt gagagtctga tgggggaggt ggtcacagac ttgaatgact cagaatgtcc
      901 agaggccaag agactgaagg tagaggagcc gggcatgggt gcagaggagc ctgaggactg
      961 tcgccagtgg acccaacacc acctggttgc tgctgatatc cgtgtggcac cagccacagc
     1021 gctgactcct cctcagggag atgcagatgc agacggagtg gatgtcaacg tccgagggcc
     1081 tgatggcttc accccactta tgctggcctc cttctgtggg ggagccctgg agccgatgcc
     1141 agctgaggag gatgaggcgg atgacacatc agccagcatt atctcagatc tgatctgtca
     1201 aggggcccag ctcggggcac ggactgaccg cactggcgag accgccctgc atttggctgc
     1261 ccgctatgct agagcggatg cagccaagcg tctcctggat gctggggcgg acaccaatgc
     1321 ccaggatcat tcgggccgca cccctctgca caccgcagtg acagctgatg cccagggtgt
     1381 cttccagatt ctcatcagga accgctccac tgacctggat gcccgaatgg cagatggctc
     1441 tactgcactg atcctggcag cccgcctggc agtggagggc atggtggaag agctcatcgc
     1501 cagccatgcc gatgtcaatg cagtggatga gcttgggaaa tctgccttac actgggctgc
     1561 agctgtgaac aacgtggagg ctaccttggc tctgctgaaa aatggagcca acaaggacat
     1621 gcaggacagc aaggaagaga cgccgctgtt cttggccgct cgggagggca gctatgaggc
     1681 tgccaagctg ctgctggatc atctcgccaa ccgggagatc acagatcact tggacaggct
     1741 gccccgggac gtggcccagg agcggctgca ccaggacatt gtgcggttgc tggaccagcc
     1801 cagtggacct cgaagtccct ctggtcccca tggcttaggg ccattgctct gcccaccagg
     1861 ggccttcctt cctggcctca aagcggtgca gtctgggacc aagaagagca ggaggccacc
     1921 tggcaagacc gggctggggc cacagggaac tcgtggtcgg ggcaagaagc tgacactagc
     1981 ctgtccagga cctctggcag acagctctgt cacactgtca ccggtggact ctctggactc
     2041 accacggcct ttcagtgggc cccctgcttc ccctggaggc ttccccttgg agggccccta
     2101 tgccaccacg gccaccgcgg tgtccttggc acagctaggc gcaagtaggg cgggtcctct
     2161 ggggcgccag cctcctgggg gctgtgtgct cagctttggt ctgctcaatc ctgtagctgt
     2221 tcccctcgac tgggccaggc tgcctccacc tgcccctcca gggccctcat tcctgctgcc
     2281 cctggctccg ggaccccagt tgctcaaccc aggagcccca gtttctcccc aagagcggcc
     2341 cccaccctac ctggctgctc caggacatgg agaggaatat cctgcagcag gaacccgcag
     2401 tagccccacc aaggcccgct tcctgcgggt tcccagcgag catccttatt tgaccccgtc
     2461 tcctgagtcc ccagagcact gggccagccc atccccccca tccctctcag actggtctga
     2521 ctcaacacct agcccagcaa ctgctaccaa tgccacagcc tctggagccc tgcctgctca
     2581 gccacacccc atatctgttc cctccctccc tcagtcccag actcagctgg gaccccaacc
     2641 agaagttacc cccaagaggc aggtgatggc ctagggatcc cgggtggcat ccctgtgacc
     2701 cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc agccttgtcc
     2761 taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat attatggggt
     2821 ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg gcctgcgggg
     2881 tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca atctccgcct
     2941 cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc caggcatgca
     3001 tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc atattggcca
     3061 ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc aaattgctgg
     3121 gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttaaaa taactatacc
     3181 agcaggagga cgtccagaca cagcataggc tacctggcca tgcccaaccg gtgggacatt
     3241 tgagttgctt gcttggcact gtcctctcat gcgttgggtc cactcagtag atgcctgttg
     3301 aattgggtac gcggccagct tggctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3361 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3421 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3481 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3541 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3601 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     3661 aaaagctcct cgaggaactg aaaaaccaga aagttaattc cctatagtga gtcgtattaa
     3721 attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     3781 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     3841 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     3901 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     3961 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     4021 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     4081 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     4141 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4201 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4261 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4321 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4381 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4441 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4501 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     4561 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     4621 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     4681 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     4741 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     4801 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     4861 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     4921 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     4981 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     5041 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     5101 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     5161 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5221 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5281 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5341 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5401 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     5461 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     5521 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     5581 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     5641 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     5701 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     5761 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     5821 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     5881 acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     5941 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     6001 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     6061 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     6121 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     6181 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     6241 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     6301 atttaacgcg aattttaaca aaatattaac gcttacaatt tccattcgcc attcaggctg
     6361 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
     6421 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     6481 tgtaaaacga cggccagtgc caagctgatc tatacattga atcaatattg gcaattagcc
     6541 atattagtca ttggttatat agcataaatc aatattggct attggccatt gcatacgttg
     6601 tatctatatc ataatatgta catttatatt ggctcatgtc caatatgacc gccatgtt
//
