LOCUS       sequence-450305-        7057 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 7057)
  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 7057)
  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: 3XFlagNICD1
FEATURES             Location/Qualifiers
     source          1..7057
                     /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    3081..3703
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3280..3299)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3727..3747)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3732..4061
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3912..4047
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4099..4118)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4100..4118)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4132..4148)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4132..4148)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4145..4167)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4156..4172
                     /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(4180..4186,4187..4204,4205..4210))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4225..4246
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4363..4380)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4534..5122)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4614..4633)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5293..6084,6085..6153))
                     /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     5916..5935
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6154..6258)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6285..6740
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6372..6391)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6582..6603
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6866..6888
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6880..6897
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6881..6897
                     /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 tctagagtgc tgctgtcccg
      721 caagcgccgg cggcagcatg gccagctctg gttccctgag ggtttcaaag tgtcagaggc
      781 cagcaagaag aagcggagag agcccctcgg cgaggactca gtcggcctca agcccctgaa
      841 gaatgcctca gatggtgctc tgatggacga caatcagaac gagtggggag acgaagacct
      901 ggagaccaag aagttccggt ttgaggagcc agtagttctc cctgacctga gtgatcagac
      961 tgaccacaga cagtggaccc agcagcacct ggacgctgct gacctgcgca tgtctgccat
     1021 ggccccaaca ccgcctcagg gggaggtgga tgctgactgc atggatgtca atgttcgagg
     1081 accagatggc ttcacacccc tcatgattgc ctcctgcagt ggagggggcc ttgagacagg
     1141 caacagtgaa gaagaagaag atgcacctgc tgtcatctct gacttcatct accagggcgc
     1201 cagcttgcac aaccagacag accgcaccgg ggagaccgcc ttgcacttgg ctgcccgata
     1261 ctctcgttca gatgctgcaa agcgcttgct ggaggccagt gcagatgcca acatccagga
     1321 caacatgggc cgtactccgt tacatgcagc agtttctgca gatgctcagg gtgtcttcca
     1381 gatcctgctc cggaacaggg ccacagatct ggatgcccga atgcatgatg gcacaactcc
     1441 actgatcctg gctgcgcgcc tggccctgga gggcatgctg gaggacctca tcaactcaca
     1501 tgctgacgtc aatgccgtgg atgacctagg caagtcggct ttgcattggg cggccgcggt
     1561 gaacaatgtg gatgctgctg ttgtgctcct gaagaacgga gccaacaagg acatgcagaa
     1621 caacaaggag gagactcccc tgttcctggc cgcccgtgag ggcagctatg agactgccaa
     1681 agtgttgctg gaccactttg ccaaccggga catcacggat cacatggacc gattgccgcg
     1741 ggacatcgca caggagcgta tgcaccacga tatcgtgcgg cttttggatg agtacaacct
     1801 ggtgcgcagc ccacagctgc atggcactgc cctgggtggc acacccactc tgtctcccac
     1861 actctgctcg cccaatggct acctgggcaa tctcaagtct gccacacagg gcaagaaggc
     1921 ccgcaagccc agcaccaaag ggctggcttg tggtagcaag gaagctaagg acctcaaggc
     1981 acggaggaag aagtcccagg atggcaaggg ctgcctgttg gacagctcga gcatgctgtc
     2041 gcctgtggac tccctcgagt caccccatgg ctacttgtca gatgtggcct cgccacccct
     2101 cctcccctcc ccattccagc agtctccatc catgcctctc agccacctgc ctggtatgcc
     2161 tgacactcac ctgggcatca gccacttgaa tgtggcagcc aagcctgaga tggcagcact
     2221 ggctggaggt agccggttgg cctttgagcc acccccgcca cgcctctccc acctgcctgt
     2281 agcctccagt gccagcacag tgctgagtac caatggcacg ggggctatga atttcaccgt
     2341 gggtgcaccg gcaagcttga atggccagtg tgagtggctt ccccggctcc agaatggcat
     2401 ggtgcccagc cagtacaacc cactacggcc gggtgtgacg ccgggcacac tgagcacaca
     2461 ggcagctggc ctccagcata gcatgatggg gccactacac agcagcctct ccaccaatac
     2521 cttgtccccg attatttacc agggcctgcc caacacacgg ctggcaacac agcctcacct
     2581 ggtgcagacc cagcaggtgc agccacagaa cttacagctc cagcctcaga acctgcagcc
     2641 accatcacag ccacacctca gtgtgagctc ggcagccaat gggcacctgg gccggagctt
     2701 cttgagtggg gagcccagtc aggcagatgt acaaccgctg ggccccagca gtctgcctgt
     2761 gcacaccatt ctgccccagg aaagccaggc cctgcccaca tcactgccat cctccatggt
     2821 cccacccatg accactaccc agttcctgac ccctccttcc cagcacagtt actcctcctc
     2881 ccctgtggac aacaccccca gccaccagct gcaggtgcca gagcacccct tcctcacccc
     2941 atcccctgag tcccctgacc agtggtccag ctcctccccg cattccaaca tctctgattg
     3001 gtccgagggc atctccagcc cgcccaccac catgccgtcc cagatcaccc acattccaga
     3061 ggcatttaaa taaggatccc gggtggcatc cctgtgaccc ctccccagtg cctctcctgg
     3121 ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta agttgcatca
     3181 ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg gtatggagca
     3241 aggggcaagt tgggaagaca acctgtaggg cctgcggggt ctattgggaa ccaagctgga
     3301 gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc
     3361 tgcctcagcc tcccgagttg ttgggattcc aggcatgcat gaccaggctc agctaatttt
     3421 tgtttttttg gtagagacgg ggtttcacca tattggccag gctggtctcc aactcctaat
     3481 ctcaggtgat ctacccacct tggcctccca aattgctggg attacaggcg tgaaccactg
     3541 ctcccttccc tgtccttctg attttaaaat aactatacca gcaggaggac gtccagacac
     3601 agcataggct acctggccat gcccaaccgg tgggacattt gagttgcttg cttggcactg
     3661 tcctctcatg cgttgggtcc actcagtaga tgcctgttga attgggtacg cggccagctt
     3721 ggctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3781 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     3841 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     3901 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     3961 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     4021 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcctc gaggaactga
     4081 aaaaccagaa agttaattcc ctatagtgag tcgtattaaa ttcgtaatca tgtcatagct
     4141 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     4201 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     4261 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     4321 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     4381 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     4441 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4501 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4561 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4621 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4681 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4741 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4801 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4861 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4921 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     4981 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5041 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     5101 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5161 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5221 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     5281 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5341 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5401 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5461 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5521 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5581 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5641 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5701 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5761 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5821 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5881 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5941 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6001 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6061 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6121 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6181 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6241 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgcgc cctgtagcgg
     6301 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     6361 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     6421 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     6481 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     6541 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     6601 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     6661 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     6721 aatattaacg cttacaattt ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     6781 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     6841 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc
     6901 aagctgatct atacattgaa tcaatattgg caattagcca tattagtcat tggttatata
     6961 gcataaatca atattggcta ttggccattg catacgttgt atctatatca taatatgtac
     7021 atttatattg gctcatgtcc aatatgaccg ccatgtt
//
