LOCUS       sequence-176181-        7651 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Myc tagged NICD1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7651)
  AUTHORS   Fan B, Malato Y, Calvisi DF, Naqvi S, Razumilava N, Ribback S, Gores
            GJ, Dombrowski F, Evert M, Chen X, Willenbring H
  TITLE     Cholangiocarcinomas can originate from hepatocytes in mice.
  JOURNAL   J Clin Invest. 2012 Aug 1;122(8):2911-5. doi: 10.1172/JCI63212. Epub
            2012 Jul 17.
  PUBMED    22797301
REFERENCE   2  (bases 1 to 7651)
  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"; doi: "10.1172/JCI63212"; 
            journalName: "J Clin Invest"; date: "2012-08-1- 1"; volume: "122"; 
            issue: "8"; pages: "2911-5"
COMMENT     Sequence Label: pT3-EF1aH NICD1
FEATURES             Location/Qualifiers
     source          1..7651
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..1179
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          232..1170
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1127..1147
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1210..1234
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1271..1300
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    complement(3758..3782)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     3810..3829
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3864..3881)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3870..4094
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4512..4528)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4512..4528)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4525..4547)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4536..4552
                     /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(4560..4566,4567..4584,4585..4590))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4605..4626
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4743..4760)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4914..5502)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4994..5013)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5673..6464,6465..6533))
                     /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     6296..6315
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6534..6638)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6706..6724
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6762..6784)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6884..6903
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7097..7119
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7111..7128
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7112..7128
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7510..7530)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
       61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
      781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
      841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
      901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
      961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gaattagctt ggtaccaata
     1201 cggatatcaa caagtttgta caaaaaagca ggctttaaag gaaccaattc agtcgactgg
     1261 atccaccatg gagcaaaagc tcatttctga agaggacttg agatccacca tgcagcatgg
     1321 ccagctctgg ttccctgagg gtttcaaagt gtcagaggcc agcaagaaga agcggagaga
     1381 gcccctcggc gaggactcag tcggcctcaa gcccctgaag aatgcctcag atggtgctct
     1441 gatggacgac aatcagaacg agtggggaga cgaagacctg gagaccaaga agttccggtt
     1501 tgaggagcca gtagttctcc ctgacctgag tgatcagact gaccacagac agtggaccca
     1561 gcagcacctg gacgctgctg acctgcgcat gtctgccatg gccccaacac cgcctcaggg
     1621 ggaggtggat gctgactgca tggatgtcaa tgttcgagga ccagatggct tcacacccct
     1681 catgattgcc tcctgcagtg gagggggcct tgagacaggc aacagtgaag aagaagaaga
     1741 tgcacctgct gtcatctctg acttcatcta ccagggcgcc agcttgcaca accagacaga
     1801 ccgcaccggg gagaccgcct tgcacttggc tgcccgatac tctcgttcag atgctgcaaa
     1861 gcgcttgctg gaggccagtg cagatgccaa catccaggac aacatgggcc gtactccgtt
     1921 acatgcagca gtttctgcag atgctcaggg tgtcttccag atcctgctcc ggaacagggc
     1981 cacagatctg gatgcccgaa tgcatgatgg cacaactcca ctgatcctgg ctgcgcgcct
     2041 ggccctggag ggcatgctgg aggacctcat caactcacat gctgacgtca atgccgtgga
     2101 tgacctaggc aagtcggctt tgcattgggc ggccgcggtg aacaatgtgg atgctgctgt
     2161 tgtgctcctg aagaacggag ccaacaagga catgcagaac aacaaggagg agactcccct
     2221 gttcctggcc gcccgtgagg gcagctatga gactgccaaa gtgttgctgg accactttgc
     2281 caaccgggac atcacggatc acatggaccg attgccgcgg gacatcgcac aggagcgtat
     2341 gcaccacgat atcgtgcggc ttttggatga gtacaacctg gtgcgcagcc cacagctgca
     2401 tggcactgcc ctgggtggca cacccactct gtctcccaca ctctgctcgc ccaatggcta
     2461 cctgggcaat ctcaagtctg ccacacaggg caagaaggcc cgcaagccca gcaccaaagg
     2521 gctggcttgt ggtagcaagg aagctaagga cctcaaggca cggaggaaga agtcccagga
     2581 tggcaagggc tgcctgttgg acagctcgag catgctgtcg cctgtggact ccctcgagtc
     2641 accccatggc tacttgtcag atgtggcctc gccacccctc ctcccctccc cattccagca
     2701 gtctccatcc atgcctctca gccacctgcc tggtatgcct gacactcacc tgggcatcag
     2761 ccacttgaat gtggcagcca agcctgagat ggcagcactg gctggaggta gccggttggc
     2821 ctttgagcca cccccgccac gcctctccca cctgcctgta gcctccagtg ccagcacagt
     2881 gctgagtacc aatggcacgg gggctatgaa tttcaccgtg ggtgcaccgg caagcttgaa
     2941 tggccagtgt gagtggcttc cccggctcca gaatggcatg gtgcccagcc agtacaaccc
     3001 actacggccg ggtgtgacgc cgggcacact gagcacacag gcagctggcc tccagcatag
     3061 catgatgggg ccactacaca gcagcctctc caccaatacc ttgtccccga ttatttacca
     3121 gggcctgccc aacacacggc tggcaacaca gcctcacctg gtgcagaccc agcaggtgca
     3181 gccacagaac ttacagctcc agcctcagaa cctgcagcca ccatcacagc cacacctcag
     3241 tgtgagctcg gcagccaatg ggcacctggg ccggagcttc ttgagtgggg agcccagtca
     3301 ggcagatgta caaccgctgg gccccagcag tctgcctgtg cacaccattc tgccccagga
     3361 aagccaggcc ctgcccacat cactgccatc ctccatggtc ccacccatga ccactaccca
     3421 gttcctgacc cctccttccc agcacagtta ctcctcctcc cctgtggaca acacccccag
     3481 ccaccagctg caggtgccag agcacccctt cctcacccca tcccctgagt cccctgacca
     3541 gtggtccagc tcctccccgc attccaacat ctctgattgg tccgagggca tctccagccc
     3601 gcccaccacc atgccgtccc agatcaccca cattccagag gcatttaaat aaacagagat
     3661 gtgggatgca ggaccccagc ttccgttccc aagccctgtt gggagtcctt tacagtgctt
     3721 caggatgcga attcgcggcc gcactcgaga tatctagacc cagctttctt gtacaaagtg
     3781 gttgatatcg cggccgctgc attctagttg tggtttgtcc aaactcatca atgtatctta
     3841 tcatgtctgg atccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     3901 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     3961 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     4021 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     4081 ggtgggctct atggcttctg aggcgggatc cccttgaaat acatccacag gtacacctcc
     4141 aattgactca aatgatgtca attagtctat cagaagcttc taaagccatg acatcatttt
     4201 ctggaatttt ccaagctgtt taaaggcaca gtcaacttag tgtatgtaaa cttctgaccc
     4261 actggaattg tgatacagtg aattataagt gaaataatct gtctgtaaac aattgttgga
     4321 aaaatgactt gtgtcatgca caaagtagat gtcctaactg acttgccaaa actattgttt
     4381 gttaacaaga aatttgtgga gtagttgaaa aacgagtttt aatgactcca acttaagtgt
     4441 atgtaaactt ccgacttcaa ctgtataggt ctagagtcga cctgcaggca tgcaagcttg
     4501 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     4561 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     4621 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     4681 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     4741 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     4801 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     4861 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4921 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4981 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5041 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5101 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5161 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5221 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5281 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5341 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5401 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5461 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     5521 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     5581 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     5641 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     5701 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     5761 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     5821 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     5881 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     5941 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     6001 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     6061 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     6121 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     6181 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     6241 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     6301 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     6361 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     6421 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     6481 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     6541 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     6601 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     6661 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     6721 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     6781 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     6841 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     6901 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     6961 cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     7021 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     7081 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga attcgagctc
     7141 ggtaccctac agttgaagtc ggaagtttac atacacttaa gttggagtca ttaaaactcg
     7201 tttttcaact actccacaaa tttcttgtta acaaacaata gttttggcaa gtcagttagg
     7261 acatctactt tgtgcatgac acaagtcatt tttccaacaa ttgtttacag acagattatt
     7321 tcacttataa ttcactgtat cacaattcca gtgggtcaga agtttacata cactaagttg
     7381 actgtgcctt taaacagctt ggaaaattcc agaaaatgat gtcatggctt tagaagcttc
     7441 tgatagacta attgacatca tttgagtcaa ttggaggtgt acctgtggat gtatttcaag
     7501 gaattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     7561 gaagtatgca aagcatgcat atcgatacta gtttaattaa gatcctcttt gcagctaatg
     7621 gaccttctag gtcttgaaag gagtgggaat t
//
