LOCUS       sequence-339584-        8856 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  full length clock in pCMV10 with 3x flag at the N-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8856)
  AUTHORS   Huang N, Chelliah Y, Shan Y, Taylor CA, Yoo SH, Partch C, Green CB, 
            Zhang H, Takahashi JS
  TITLE     Crystal structure of the heterodimeric CLOCK:BMAL1 transcriptional 
            activator complex.
  JOURNAL   Science. 2012 Jul 13;337(6091):189-94. doi: 10.1126/science.1222804.
            Epub 2012 May 31.
  PUBMED    22653727
REFERENCE   2  (bases 1 to 8856)
  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: 
            "10.1126/science.1222804"; journalName: "Science"; date: 
            "2012-07-13- 13"; volume: "337"; issue: "6091"; pages: "189-94"
COMMENT     Sequence Label: pCMV10/3xflag-Clock
FEATURES             Location/Qualifiers
     source          1..8856
                     /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    3298..3920
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3497..3516)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3944..3964)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3962..4259
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4313..5107
                     /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(4367..4386)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4977..4996
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5759..5893
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5796..5815)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5850..5869
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5898..5917)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5899..5917)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5931..5947)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5931..5947)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5944..5966)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5955..5971
                     /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(5979..5985,5986..6003,6004..6009))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6024..6045
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6162..6179)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6333..6921)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6413..6432)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7092..7883,7884..7952))
                     /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     7715..7734
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7953..8057)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      8084..8539
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8171..8190)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8381..8402
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8665..8687
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8679..8696
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8680..8696
                     /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 agaattaacc atggactaca aagaccatga cggtgattat aaagatcatg acatcgatta
      661 caaggatgac gatgacaagc ttgcggccgc gatggtgttt accgtaagct gtagtaaaat
      721 gagctcaatt gttgacagag atgacagtag tatttttgat ggattggtgg aagaagatga
      781 caaggacaaa gcaaaaagag tatctagaaa caaatcagaa aagaaacgta gagatcagtt
      841 caatgtcctc attaaggagc tggggtctat gcttcctggt aacgcgagaa agatggacaa
      901 gtctactgtt ctacagaaga gcattgattt tttgcgcaaa cataaagaga ccactgcaca
      961 gtcagatgct agtgagattc gacaggactg gaaacccaca ttccttagta atgaagagtt
     1021 tacacagtta atgttagagg ctcttgatgg ttttttttta gcgatcatga cagatggaag
     1081 tataatatat gtatctgaga gtgtaacttc gttacttgaa catttaccat ctgatcttgt
     1141 ggatcaaagt atatttaatt ttatcccaga gggagaacat tcagaggttt ataagatact
     1201 ctctactcat ctgctggaaa gtgactcatt aacccctgag tacttaaaat caaaaaatca
     1261 gttagaattc tgttgtcaca tgcttcgagg aacaatagac ccaaaggagc catccaccta
     1321 tgaatatgtg agatttatag gaaattttaa atctttaacc agtgtatcaa cttcaacaca
     1381 caatggtttt gaaggaacta tacaacgcac acataggcct tcttatgaag atagagtttg
     1441 ttttgtagct actgtcagat tagctacacc tcagttcatc aaggaaatgt gtactgttga
     1501 agaaccaaat gaagagttta catctagaca cagtttagaa tggaagtttc tatttttaga
     1561 tcacagggca ccaccaataa taggctattt gccatttgaa gtcttgggaa catcaggcta
     1621 tgattactat catgtggatg acctagaaaa tctggcaaaa tgtcacgagc acttaatgca
     1681 atatggaaaa ggcaaatcgt gttactatag attcctgacc aaaggccagc agtggatatg
     1741 gcttcagact cattattata ttacttacca tcagtggaat tcaaggccag agttcattgt
     1801 ttgtactcac actgtagtaa gttatgcaga agttagggct gaaagacggc gagaacttgg
     1861 cattgaagag tctcttcctg agacagctgc tgacaaaagc caagattctg ggtctgacaa
     1921 tcgtatcaac acagtgagtc tcaaggaagc actggaaagg tttgatcaca gcccaactcc
     1981 ttctgcctcc tctagaagct cacgaaagtc atctcacacc gcagtctcag acccttcctc
     2041 cacaccgaca aagatcccta ctgatactag cactcctccc agacagcatt tgccagctca
     2101 tgaaaagatg acacagcgga ggtcgtcctt cagcagtcag tccataaact cccagtcagt
     2161 tggtccatca ttaacacagc cagcgatgtc tcaagctgca aatttaccaa ttccacaagg
     2221 catgtcacag tttcagtttt cagctcagtt aggagccatg cagcatctaa aagaccagct
     2281 agagcagcgg acacggatga tagaggcaaa tattcatcgg cagcaagaag aactaaggaa
     2341 aattcaagag caacttcaga tggtccatgg tcaagggcta cagatgtttt tgcagcaatc
     2401 aaaccctgga ttgaattttg gttctgttca actttcctct ggaaattcta atatccagca
     2461 actcacacct gtaaatatgc aaggccaggt tgtccctgct aaccaggttc agagtggaca
     2521 tatcagcaca ggccagcaca tgatacagca acagacttta caaagtacat caactcagca
     2581 gagtcaacag agtgtaatga gtggacacag tcagcagacg tctcttccaa gtcagacacc
     2641 gagcactctc acagccccac tgtacaatac gatggtgatt tcccagcctg cagctgggag
     2701 catggtccag attccatcca gtatgccaca gaacagtacc cagagtgcta cagtcactac
     2761 gttcactcag gacagacaga taagattttc tcaaggtcag caacttgtga ccaaattagt
     2821 gactgctcct gtagcttgtg gggccgtcat ggtaccaagt accatgctta tgggtcaggt
     2881 ggtgactgcc tatcctacct tcgccacaca acagcagcag gcacagacat tatcggtaac
     2941 acaacagcag cagcagcagc agcagcagcc accacagcaa cagcaacaac aacagcagag
     3001 ttcccaggaa cagcagcttc cttcagttca gcagccagct caggcccagc tgggccagcc
     3061 accacagcag ttcttacaga catctaggtt gctccacggg aatccttcga cacagctcat
     3121 cctctctgct gcctttccac tacaacagag cactttccct ccttcgcacc accagcaaca
     3181 ccagcctcag cagcaacagc agcttcctcg gcacaggact gacagcctga ctgacccttc
     3241 caaggtccag ccacagtaga gatctgatat cggtaccagt cgactctaga ggatcccggg
     3301 tggcatccct gtgacccctc cccagtgcct ctcctggccc tggaagttgc cactccagtg
     3361 cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc
     3421 tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc
     3481 tgtagggcct gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc
     3541 actgcaatct ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc cgagttgttg
     3601 ggattccagg catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt
     3661 ttcaccatat tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg
     3721 cctcccaaat tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgatt
     3781 ttaaaataac tataccagca ggaggacgtc cagacacagc ataggctacc tggccatgcc
     3841 caaccggtgg gacatttgag ttgcttgctt ggcactgtcc tctcatgcgt tgggtccact
     3901 cagtagatgc ctgttgaatt gggtacgcgg ccagcttggc tgtggaatgt gtgtcagtta
     3961 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4021 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4081 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4141 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4201 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4261 ggaattgatc agcttgggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     4321 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     4381 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     4441 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     4501 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     4561 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     4621 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     4681 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     4741 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     4801 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     4861 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     4921 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     4981 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     5041 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     5101 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     5161 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     5221 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     5281 gggctcgatc ccctcgcgag ttggttcagc tgctgcctga ggctggacga cctcgcggag
     5341 ttctaccggc agtgcaaatc cgtcggcatc caggaaacca gcagcggcta tccgcgcatc
     5401 catgcccccg aactgcagga gtggggaggc acgatggccg ctttggtcga cccggacggg
     5461 acgctcctgc gcctgataca gaacgaattg cttgcaggca tctcatgagt gtgtcttccc
     5521 gttttccgcc tgaggtcact gcgtggatgg agcgctggcg cctgctgcgc gacggcgagc
     5581 tgctcaccac ccactcgcca agctggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     5641 ataggctccg ccgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     5701 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     5761 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     5821 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     5881 tcatgtctgg atcaattccc tatagtgagt cgtattaaat tcgtaatcat gtcatagctg
     5941 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     6001 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     6061 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     6121 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     6181 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     6241 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     6301 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     6361 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     6421 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     6481 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     6541 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     6601 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     6661 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     6721 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     6781 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     6841 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     6901 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     6961 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     7021 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     7081 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     7141 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     7201 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     7261 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     7321 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     7381 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     7441 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     7501 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     7561 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     7621 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     7681 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     7741 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     7801 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     7861 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     7921 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     7981 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     8041 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgcgcc ctgtagcggc
     8101 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     8161 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     8221 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     8281 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     8341 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     8401 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     8461 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     8521 atattaacgc ttacaatttc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     8581 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     8641 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca
     8701 agctgatcta tacattgaat caatattggc aattagccat attagtcatt ggttatatag
     8761 cataaatcaa tattggctat tggccattgc atacgttgta tctatatcat aatatgtaca
     8821 tttatattgg ctcatgtcca atatgaccgc catgtt
//
