LOCUS       sequence-115728-        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"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     235..254
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        235..253
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(292..621)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      306..441
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     606..626
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    650..1272
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     1054..1073
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     CDS             complement(3674..3739)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     promoter        complement(3769..3972)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3798..3818)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3973..4352
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(4513..4530)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4513..4529)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4522..4544)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(4670..5125)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4807..4828)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5019..5038
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5152..5256
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5257..5325,5326..6117)
                     /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     complement(5475..5494)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6288..6876
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6777..6796
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7030..7047
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga catgattacg aatttaatac
      241 gactcactat agggaattaa ctttctggtt tttcagttcc tcgaggagct ttttgcaaaa
      301 gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc
      361 ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact
      421 gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt
      481 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt
      541 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
      601 tccacaccct aactgacaca cattccacag ccaagctggc cgcgtaccca attcaacagg
      661 catctactga gtggacccaa cgcatgagag gacagtgcca agcaagcaac tcaaatgtcc
      721 caccggttgg gcatggccag gtagcctatg ctgtgtctgg acgtcctcct gctggtatag
      781 ttattttaaa atcagaagga cagggaaggg agcagtggtt cacgcctgta atcccagcaa
      841 tttgggaggc caaggtgggt agatcacctg agattaggag ttggagacca gcctggccaa
      901 tatggtgaaa ccccgtctct accaaaaaaa caaaaattag ctgagcctgg tcatgcatgc
      961 ctggaatccc aacaactcgg gaggctgagg caggagaatc gcttgaaccc aggaggcgga
     1021 gattgcagtg agccaagatt gtgccactgc actccagctt ggttcccaat agaccccgca
     1081 ggccctacag gttgtcttcc caacttgccc cttgctccat accacccccc tccaccccat
     1141 aatattatag aaggacacct agtcagacaa aatgatgcaa cttaatttta ttaggacaag
     1201 gctggtgggc actggagtgg caacttccag ggccaggaga ggcactgggg aggggtcaca
     1261 gggatgccac ccgggatcct tatttaaatg cctctggaat gtgggtgatc tgggacggca
     1321 tggtggtggg cgggctggag atgccctcgg accaatcaga gatgttggaa tgcggggagg
     1381 agctggacca ctggtcaggg gactcagggg atggggtgag gaaggggtgc tctggcacct
     1441 gcagctggtg gctgggggtg ttgtccacag gggaggagga gtaactgtgc tgggaaggag
     1501 gggtcaggaa ctgggtagtg gtcatgggtg ggaccatgga ggatggcagt gatgtgggca
     1561 gggcctggct ttcctggggc agaatggtgt gcacaggcag actgctgggg cccagcggtt
     1621 gtacatctgc ctgactgggc tccccactca agaagctccg gcccaggtgc ccattggctg
     1681 ccgagctcac actgaggtgt ggctgtgatg gtggctgcag gttctgaggc tggagctgta
     1741 agttctgtgg ctgcacctgc tgggtctgca ccaggtgagg ctgtgttgcc agccgtgtgt
     1801 tgggcaggcc ctggtaaata atcggggaca aggtattggt ggagaggctg ctgtgtagtg
     1861 gccccatcat gctatgctgg aggccagctg cctgtgtgct cagtgtgccc ggcgtcacac
     1921 ccggccgtag tgggttgtac tggctgggca ccatgccatt ctggagccgg ggaagccact
     1981 cacactggcc attcaagctt gccggtgcac ccacggtgaa attcatagcc cccgtgccat
     2041 tggtactcag cactgtgctg gcactggagg ctacaggcag gtgggagagg cgtggcgggg
     2101 gtggctcaaa ggccaaccgg ctacctccag ccagtgctgc catctcaggc ttggctgcca
     2161 cattcaagtg gctgatgccc aggtgagtgt caggcatacc aggcaggtgg ctgagaggca
     2221 tggatggaga ctgctggaat ggggagggga ggaggggtgg cgaggccaca tctgacaagt
     2281 agccatgggg tgactcgagg gagtccacag gcgacagcat gctcgagctg tccaacaggc
     2341 agcccttgcc atcctgggac ttcttcctcc gtgccttgag gtccttagct tccttgctac
     2401 cacaagccag ccctttggtg ctgggcttgc gggccttctt gccctgtgtg gcagacttga
     2461 gattgcccag gtagccattg ggcgagcaga gtgtgggaga cagagtgggt gtgccaccca
     2521 gggcagtgcc atgcagctgt gggctgcgca ccaggttgta ctcatccaaa agccgcacga
     2581 tatcgtggtg catacgctcc tgtgcgatgt cccgcggcaa tcggtccatg tgatccgtga
     2641 tgtcccggtt ggcaaagtgg tccagcaaca ctttggcagt ctcatagctg ccctcacggg
     2701 cggccaggaa caggggagtc tcctccttgt tgttctgcat gtccttgttg gctccgttct
     2761 tcaggagcac aacagcagca tccacattgt tcaccgcggc cgcccaatgc aaagccgact
     2821 tgcctaggtc atccacggca ttgacgtcag catgtgagtt gatgaggtcc tccagcatgc
     2881 cctccagggc caggcgcgca gccaggatca gtggagttgt gccatcatgc attcgggcat
     2941 ccagatctgt ggccctgttc cggagcagga tctggaagac accctgagca tctgcagaaa
     3001 ctgctgcatg taacggagta cggcccatgt tgtcctggat gttggcatct gcactggcct
     3061 ccagcaagcg ctttgcagca tctgaacgag agtatcgggc agccaagtgc aaggcggtct
     3121 ccccggtgcg gtctgtctgg ttgtgcaagc tggcgccctg gtagatgaag tcagagatga
     3181 cagcaggtgc atcttcttct tcttcactgt tgcctgtctc aaggccccct ccactgcagg
     3241 aggcaatcat gaggggtgtg aagccatctg gtcctcgaac attgacatcc atgcagtcag
     3301 catccacctc cccctgaggc ggtgttgggg ccatggcaga catgcgcagg tcagcagcgt
     3361 ccaggtgctg ctgggtccac tgtctgtggt cagtctgatc actcaggtca gggagaacta
     3421 ctggctcctc aaaccggaac ttcttggtct ccaggtcttc gtctccccac tcgttctgat
     3481 tgtcgtccat cagagcacca tctgaggcat tcttcagggg cttgaggccg actgagtcct
     3541 cgccgagggg ctctctccgc ttcttcttgc tggcctctga cactttgaaa ccctcaggga
     3601 accagagctg gccatgctgc cgccggcgct tgcgggacag cagcactcta gagtcgacct
     3661 gcaggcatgc aagcttgtca tcgtcatcct tgtaatcgat atcatgatct ttataatcac
     3721 cgtcatggtc tttgtagtcc atggtgaatt ctgacggttc actaaacgag ctctgcttat
     3781 atagacctcc caccgtacac gcctaccgcc catttgcgtc aacggggcgg ggttattacg
     3841 acattttgga aagtcccgtt gattttggtg ccaaaacaaa ctcccattga cgtcaatggg
     3901 gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca cgcccattgg tgtactgcca
     3961 aaaccgcatc accatggtaa tagcgatgac taatacgtag atgtactgcc aagtaggaaa
     4021 gtcccgtaag gtcatgtact gggcataatg ccaggcgggc catttaccgt cattgacgtc
     4081 aatagggggc ggacttggca tatgatacac ttgatgtact gccaagtggg cagtttaccg
     4141 taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta tgggaacata
     4201 cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg ggccatttac
     4261 cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc ccgtaattga
     4321 ttactattaa taactagtca ataatcaatg tcaacatggc ggtcatattg gacatgagcc
     4381 aatataaatg tacatattat gatatagata caacgtatgc aatggccaat agccaatatt
     4441 gatttatgct atataaccaa tgactaatat ggctaattgc caatattgat tcaatgtata
     4501 gatcagcttg gcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     4561 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     4621 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgga aattgtaagc
     4681 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     4741 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     4801 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     4861 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     4921 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     4981 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     5041 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     5101 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     5161 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     5221 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     5281 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     5341 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     5401 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     5461 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     5521 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     5581 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     5641 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     5701 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     5761 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     5821 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     5881 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     5941 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     6001 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     6061 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     6121 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     6181 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     6241 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     6301 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     6361 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     6421 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     6481 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     6541 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     6601 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     6661 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     6721 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     6781 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     6841 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     6901 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     6961 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     7021 accgagcgca gcgagtcagt gagcgaggaa gcggaag
//
