LOCUS       sequence-219884-        7012 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 7012)
  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 7012)
  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: 3XFlagNICD2
FEATURES             Location/Qualifiers
     source          1..7012
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        42..421
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        422..625
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     576..596
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             655..720
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    3077..3699
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3276..3295)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3723..3743)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3728..4057
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3908..4043
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4095..4114)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4096..4114)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4128..4144)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4128..4144)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4141..4163)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4152..4168
                     /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(4176..4182,4183..4200,4201..4206))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4221..4242
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4359..4376)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4530..5118)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4610..4629)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5289..6080,6081..6149))
                     /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     5912..5931
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6150..6254)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6281..6736
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6368..6387)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6578..6599
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6862..6884
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6876..6893
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6877..6893
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gtacatttat attggctcat gtccaatatg accgccatgt tgacattgat tattgactag
       61 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
      121 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      181 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      241 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      301 tccgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
      361 gaccttacgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
      421 ggtgatgcgg ttttggcagt acaccaatgg gcgtggatag cggtttgact cacggggatt
      481 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
      541 ctttccaaaa tgtcgtaata accccgcccc gttgacgcaa atgggcggta ggcgtgtacg
      601 gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagaattcac catggactac
      661 aaagaccatg acggtgatta taaagatcat gatatcgatt acaaggatga cgatgacaag
      721 cttgcatgcc tgcaggtcga ctctagaatg gccaagcgga agcgcaagca tggcttcctc
      781 tggctgccgg aagggttcac cctccgccga gactctagca atcacaagcg ccgtgaacct
      841 gtgggacagg atgccgtggg gctgaaaaat ctctccgtgc aagtgtcaga ggctaacctg
      901 attggttctg ggacaagtga acattgggtt gatgatgaag gaccccagcc aaagaaagcc
      961 aaggctgagg atgaggcttt gctgtcggaa gatgacccca tcgatcgacg gccctggaca
     1021 cagcagcacc ttgaagctgc agacatccgc cacactccat ccctggcact cactcctcct
     1081 caggcagaac aggaggtgga cgtgctggac gtgaatgtcc gaggcccaga tgggtgtact
     1141 ccactgatgc tggcttctct ccgaggaggc agctcagacc tgagtgatga agacgaagat
     1201 gctgaggact cttctgccaa catcatcaca gacttggtct accaaggtgc cagccttcaa
     1261 gcacagacag accgcactgg cgagatggcc ctgcaccttg cagcccgcta ttcgagagct
     1321 gatgctgcca aacgcctcct ggatgctggt gcggatgcaa atgcccagga caacatgggc
     1381 cgatgtcctc ttcacgctgc ggtggcagca gacgcccaag gtgtctttca gattctgatc
     1441 cgcaaccgtg taaccgatct ggatgccaga atgaacgatg gtactacccc cctgatcctg
     1501 gctgcccgcc tggctgtgga aggaatggtg gcagagttga tcaattgcca agcagatgtc
     1561 aatgcagtgg atgatcatgg aaaatctgcc ctccactggg cagctgctgt caataatgtg
     1621 gaggcgactc ttctgctgtt gaagaatggg gccaacagag atatgcagga caataaggaa
     1681 gagacacctt tgtttcttgc tgcccgagag ggaagttatg aagcggccaa aatcctgtta
     1741 gaccattttg ccaaccggga catcactgac cacatggacc gccttccccg ggatgtggct
     1801 cgggaccgca tgcaccatga catcgttcgc ctcctggacg agtataacgt gactcccagc
     1861 cctccgggaa cggtcttgac ttctgcgctc tcacctgtcc tctgtgggcc caacaggtct
     1921 ttcctcagtc tgaagcacac cccaatgggt aagaaggcta ggcggcccaa caccaagagc
     1981 accatgccca cgagcctgcc taaccttgcc aaggaggcca aggatgccaa gggcagcagg
     2041 aggaagaagt gtctgaacga gaaggtccag ctgtccgaga gctcagtgac tctatccccc
     2101 gtcgattcgc tcgagtctcc tcacacgtat gtctccgatg ccacatcctc tcccatgatc
     2161 acatcccctg gaatcttaca ggcctcgccc acccccctgc tggctgctgc cgccccggct
     2221 gccccagtgc acacacagca tgcgctgtct ttctctaacc ttcatgacat gcagcctttg
     2281 gctcctggag ccagcaccgt gctcccctcg gtcagccagc tgctatccca ccaccacatc
     2341 gcgcccccag gtagtagcag tgcaggaagc ttgggcaggt tacatccagt tcctgtccca
     2401 gcagactgga tgaaccgtgt ggagatgaac gagacccagt acagtgaaat gtttggcatg
     2461 gtcctggctc ctgcagaggg agcccaccct ggcatagcag ctccccagag cagacctccg
     2521 gaagggaagc acatgtccac ccagcgggag cccttgcctc ccatcgtgac tttccagctt
     2581 atcccaaaag gcagcattgc ccaggcagcc ggagctcccc agacgcagtc cagttgccct
     2641 ccagctgttg caggcccctt gccctctatg taccagatcc cagagatgcc ccgtttgccc
     2701 agtgtggctt tcccacctac catgatgccc cagcaggagg ggcaggtagc tcagaccatt
     2761 gtgccaacct atcatccttt cccagcctct gtgggcaagt accccacacc cccttcccaa
     2821 cacagttacg cctcctcaaa tgctgctgag cgaaccccca gtcatggtgg tcacctccag
     2881 ggcgagcacc catacctgac accatcccca gagtctcctg accaatggtc aagctcttca
     2941 ccacactctg catctgactg gtcagatgtg accaccagcc caactcctgg aggtggtgga
     3001 ggcggtcagc ggggacccgg aacacacatg tccgagccac cacacagcaa catgcaggtg
     3061 tatgcatagg gatcccgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct
     3121 ggaagttgcc actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt
     3181 gtctgactag gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg
     3241 gcaagttggg aagacaacct gtagggcctg cggggtctat tgggaaccaa gctggagtgc
     3301 agtggcacaa tcttggctca ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc
     3361 tcagcctccc gagttgttgg gattccaggc atgcatgacc aggctcagct aatttttgtt
     3421 tttttggtag agacggggtt tcaccatatt ggccaggctg gtctccaact cctaatctca
     3481 ggtgatctac ccaccttggc ctcccaaatt gctgggatta caggcgtgaa ccactgctcc
     3541 cttccctgtc cttctgattt taaaataact ataccagcag gaggacgtcc agacacagca
     3601 taggctacct ggccatgccc aaccggtggg acatttgagt tgcttgcttg gcactgtcct
     3661 ctcatgcgtt gggtccactc agtagatgcc tgttgaattg ggtacgcggc cagcttggct
     3721 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3781 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     3841 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     3901 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     3961 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     4021 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcctcgagg aactgaaaaa
     4081 ccagaaagtt aattccctat agtgagtcgt attaaattcg taatcatgtc atagctgttt
     4141 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     4201 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     4261 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     4321 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     4381 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     4441 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4501 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4561 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     4621 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4681 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     4741 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4801 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4861 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4921 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     4981 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     5041 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     5101 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5161 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     5221 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     5281 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     5341 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     5401 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     5461 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     5521 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     5581 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     5641 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     5701 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     5761 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     5821 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     5881 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     5941 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     6001 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     6061 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     6121 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     6181 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     6241 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca
     6301 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     6361 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     6421 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     6481 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     6541 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     6601 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     6661 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     6721 ttaacgctta caatttccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     6781 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     6841 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgccaagc
     6901 tgatctatac attgaatcaa tattggcaat tagccatatt agtcattggt tatatagcat
     6961 aaatcaatat tggctattgg ccattgcata cgttgtatct atatcataat at
//
