LOCUS       sequence-208856-        4675 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 4675)
  AUTHORS   Murtaugh LC, Stanger BZ, Kwan KM, Melton DA
  TITLE     Notch signaling controls multiple steps of pancreatic 
            differentiation.
  JOURNAL   Proc Natl Acad Sci U S A. 2003 Dec 9. 100(25):14920-5.
  PUBMED    14657333
REFERENCE   2  (bases 1 to 4675)
  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. 2003 Dec 9. 100(25):14920-5."
COMMENT     Sequence Label: pBS mNotch1-IC (DM#270)
FEATURES             Location/Qualifiers
     source          1..4675
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     98..115
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    232..253
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(268..273,274..291,292..298)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    306..322
                     /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     311..333
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     330..346
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     330..346
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     365..385
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        367..385
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     414..430
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     415..431
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             480..503
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2179..2195)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2179..2195)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2227..2246)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2228..2246)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2256..2273)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2256..2272)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2265..2287)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2414..2869
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2501..2520)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2711..2732
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2895..2999
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3000..3068,3069..3860)
                     /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(3218..3237)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4031..4619
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4520..4539
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
       61 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
      121 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
      181 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
      241 ttagctcact cattaggcac cccaggcttt acactttatg ctcccggctc gtatgttgtg
      301 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
      361 gcgcgcaatt aaccctcact aaagggaaca aaagctgggt accgggcccc ccctcgaggt
      421 cgacggtatc gataagcttg ataattctag ctagctagcc ggcccaattc ggtaccatgg
      481 actacaaaga cgatgacgat aaagcccggc ggcagcatgg ccagctctgg ttccctgagg
      541 gtttcaaagt gtcagaggcc agcaagaaga agcggagaga gcccctcggc gaggactcag
      601 tcggcctcaa gcccctgaag aatgcctcag atggtgctct gatggacgac aatcagaacg
      661 agtggggaga cgaagacctg gagaccaaga agttccggtt tgaggagcca gtagttctcc
      721 ctgacctgag tgatcagact gaccacagac agtggaccca gcagcacctg gacgctgctg
      781 acctgcgcat gtctgccatg gccccaacac cgcctcaggg ggaggtggat gctgactgca
      841 tggatgtcaa tgttcgagga ccagatggct tcacacccct catgattgcc tcctgcagtg
      901 gagggggcct tgagacaggc aacagtgaag aagaagaaga tgcacctgct gtcatctctg
      961 acttcatcta ccagggcgcc agcttgcaca accagacaga ccgcaccggg gagaccgcct
     1021 tgcacttggc tgcccgatac tctcgttcag atgctgcaaa gcgcttgctg gaggccagtg
     1081 cagatgccaa catccaggac aacatgggcc gtactccgtt acatgcagca gtttctgcag
     1141 atgctcaggg tgtcttccag atcctgctcc ggaacagggc cacagatctg gatgcccgaa
     1201 tgcatgatgg cacaactcca ctgatcctgg ctgcgcgcct ggccctggag ggcatgctgg
     1261 aggacctcat caactcacat gctgacgtca atgccgtgga tgacctaggc aagtcggctt
     1321 tgcattgggc ggccgcggtg aacaatgtgg atgctgctgt tgtgctcctg aagaacggag
     1381 ccaacaagga catgcagaac aacaaggagg agactcccct gttcctggcc gcccgtgagg
     1441 gcagctatga gactgccaaa gtgttgctgg accactttgc caaccgggac atcacggatc
     1501 acatggaccg attgccgcgg gacatcgcac aggagcgtat gcaccacgat atcgtgcggc
     1561 ttttggatga gtacaacctg gtgcgcagcc cacagctgca tggcactgcc ctgggtggca
     1621 cacccactct gtctcccaca ctctgctcgc ccaatggcta cctgggcaat ctcaagtctg
     1681 ccacacaggg caagaaggcc cgcaagccca gcaccaaagg gctggcttgt ggtagcaagg
     1741 aagctaagga cctcaaggca cggaggaaga agtcccagga tggcaagggc tgcctgttgg
     1801 acagctcgag catgctgtcg cctgtggact ccctcgagtc accccatggc tacttgtcag
     1861 atgtggcctc gccacccctc ctcccctccc cattccagca gtctccatcc atgcctctca
     1921 gccacctgcc tggtatgcct gacactcacc tgggcatcag ccacttgaat gtggcagcca
     1981 agcctgagat ggcagcactg gctggaggta gccggttggc ctttgagcca cccccgccac
     2041 gcctctccca cctgcctgta gcctccagtg ccagcacagt gctgagtacc aatggcacgg
     2101 gggctatgaa tttcaccgtg ggtgcaccgg caagctaggt agctagctcg agaattatcg
     2161 aattcctgca gcccggggga tccactagtt ctagagcggc cgccaccgcg gtggagctcc
     2221 aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt
     2281 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     2341 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     2401 aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     2461 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     2521 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     2581 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     2641 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     2701 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     2761 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     2821 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt
     2881 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     2941 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     3001 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     3061 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     3121 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     3181 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     3241 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     3301 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     3361 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     3421 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     3481 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     3541 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     3601 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     3661 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     3721 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     3781 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     3841 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     3901 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     3961 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     4021 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4081 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     4141 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     4201 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     4261 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     4321 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     4381 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     4441 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     4501 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     4561 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     4621 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctca
//
