LOCUS       sequence-210915-        6697 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human BHLHE40 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6697)
  AUTHORS   Kim J, D'Annibale S, Magliozzi R, Low TY, Jansen P, Shaltiel IA, 
            Mohammed S, Heck AJ, Medema RH, Guardavaccaro D
  TITLE     USP17- and SCFbetaTrCP--regulated degradation of DEC1 controls the 
            DNA damage response.
  JOURNAL   Mol Cell Biol. 2014 Nov 15;34(22):4177-85. doi: 
            10.1128/MCB.00530-14. Epub 2014 Sep 8.
  PUBMED    25202122
REFERENCE   2  (bases 1 to 6697)
  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: 
            "10.1128/MCB.00530-14"; journalName: "Mol Cell Biol"; date: 
            "2014-11-15- 15"; volume: "34"; issue: "22"; pages: "4177-85"
COMMENT     Sequence Label: pcDNA3-HA-BHLHE40
FEATURES             Location/Qualifiers
     source          1..6697
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(66..85)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        257..636
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        637..840
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     791..811
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     885..904
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        885..903
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             975..1001
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             975..1001
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(2272..2290)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2273..2290)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2310..2327)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2316..2540
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2586..3014
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2673..2692)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2883..2904
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3023..3043)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3028..3357
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3208..3343
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3270..3289
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3424..4218
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3478..3497)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4088..4107
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4392..4513
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4429..4448)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4483..4502
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4562..4578)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4562..4578)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4575..4597)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4586..4602
                     /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(4610..4616,4617..4634,4635..4640))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4655..4676
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4793..4810)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4964..5552)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5044..5063)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5723..6514,6515..6583))
                     /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     6346..6365
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6584..6688)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact
       61 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
      121 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
      181 gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg
      241 gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg
      301 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
      361 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
      421 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
      481 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
      541 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
      601 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
      661 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
      721 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
      781 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
      841 ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac gactcactat
      901 agggagaccc aagcttggta ccgagctcgg atccactagt aacggccgcc agtgtgctgg
      961 aattcgccac catgtacccc tacgacgtgc ccgactacgc cgagcggatc cccagcgcgc
     1021 aaccaccccc cgcctgcctg cccaaagcac cgggactgga gcacggagac ctaccaggga
     1081 tgtaccctgc ccacatgtac caagtgtaca agtcaagacg gggaataaag cggagcgagg
     1141 acagcaagga gacctacaaa ttgccgcacc ggctcatcga gaaaaagaga cgtgaccgga
     1201 ttaacgagtg catcgcccag ctgaaggatc tcctacccga acatctcaaa cttacaactt
     1261 tgggtcactt ggaaaaagca gtggttcttg aacttacctt gaagcatgtg aaagcactaa
     1321 caaacctaat tgatcagcag cagcagaaaa tcattgccct gcagagtggt ttacaagctg
     1381 gtgagctgtc agggagaaat gtcgaaacag gtcaagagat gttctgctca ggtttccaga
     1441 catgtgcccg ggaggtgctt cagtatctgg ccaagcacga gaacactcgg gacctgaagt
     1501 cttcgcagct tgtcacccac ctccaccggg tggtctcgga gctgctgcag ggtggtacct
     1561 ccaggaagcc atcagaccca gctcccaaag tgatggactt caaggaaaaa cccagctctc
     1621 cggccaaagg ttcggaaggt cctgggaaaa actgcgtgcc agtcatccag cggactttcg
     1681 ctcactcgag tggggagcag agcggcagcg acacggacac agacagtggc tatggaggag
     1741 aatcggagaa gggcgacttg cgcagtgagc agccgtgctt caaaagtgac cacggacgca
     1801 ggttcacgat gggagaaagg atcggcgcaa ttaagcaaga gtccgaagaa ccccccacaa
     1861 aaaagaaccg gatgcagctt tcggatgatg aaggccattt cactagcagt gacctgatca
     1921 gctccccgtt cctgggccca cacccacacc agcctccttt ctgcctgccc ttctacctga
     1981 tcccaccttc agcgactgcc tacctgccca tgctggagaa gtgctggtat cccacctcag
     2041 tgccagtgct atacccaggc ctcaacgcct ctgccgcagc cctctctagc ttcatgaacc
     2101 cagacaagat ctcggctccc ttgctcatgc cccagagact cccttctccc ttgccagctc
     2161 atccgtccgt cgactcttct gtcttgctcc aagctctgaa gccaatcccc cctttaaact
     2221 tagaaaccaa agactaagcg gccgctcgag catgcatcta gagggcccta ttctatagtg
     2281 tcacctaaat gctagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     2341 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     2401 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     2461 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     2521 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     2581 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     2641 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     2701 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     2761 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     2821 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     2881 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     2941 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     3001 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     3061 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     3121 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3181 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     3241 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     3301 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     3361 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     3421 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     3481 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     3541 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     3601 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     3661 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     3721 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     3781 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     3841 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     3901 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     3961 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     4021 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     4081 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     4141 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     4201 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     4261 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     4321 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     4381 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     4441 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     4501 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     4561 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     4621 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     4681 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     4741 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     4801 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     4861 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     4921 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4981 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     5041 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     5101 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     5161 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     5221 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     5281 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     5341 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     5401 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     5461 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     5521 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     5581 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     5641 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     5701 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     5761 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     5821 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     5881 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     5941 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     6001 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     6061 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     6121 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     6181 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     6241 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     6301 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     6361 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     6421 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     6481 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     6541 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     6601 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     6661 agaaaaataa acaaataggg gttccgcgca catttcc
//
