LOCUS       sequence-353745-        8783 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of mutant mouse clock.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8783)
  AUTHORS   King DP, Zhao Y, Sangoram AM, Wilsbacher LD, Tanaka M, Antoch MP, 
            Steeves TD, Vitaterna MH, Kornhauser JM, Lowrey PL, Turek FW, 
            Takahashi JS
  TITLE     Positional cloning of the mouse circadian clock gene.
  JOURNAL   Cell. 1997 May 16;89(4):641-53. doi: 10.1016/s0092-8674(00)80245-7.
  PUBMED    9160755
REFERENCE   2  (bases 1 to 8783)
  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"; journalName: 
            "Cell"; date: "1997-05-16- 16"; volume: "89"; issue: "4"; pages: 
            "641-53"
COMMENT     Sequence Label: mouse Clock delta 19
FEATURES             Location/Qualifiers
     source          1..8783
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(83..103)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        88..417
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      268..403
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     330..349
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             484..1278
                     /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(538..557)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1148..1167
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1452..1573
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1489..1508)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1543..1562
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1622..1638)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1622..1638)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1635..1657)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1646..1662
                     /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(1670..1676,1677..1694,1695..1700))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1715..1736
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1853..1870)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2024..2612)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2104..2123)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2783..3574,3575..3643))
                     /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     3406..3425
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3644..3748)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3823..3842)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4014..4393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4394..4597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4548..4568
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4642..4661
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4642..4660
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(8115..8133)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(8116..8133)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(8153..8170)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8159..8383
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(8429..8783,1..74)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8516..8535)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8726..8747
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
       61 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
      121 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
      181 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
      241 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
      301 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
      361 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
      421 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
      481 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
      541 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
      601 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
      661 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
      721 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
      781 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
      841 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
      901 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
      961 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     1021 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     1081 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     1141 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     1201 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     1261 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     1321 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     1381 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     1441 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     1501 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     1561 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     1621 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     1681 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     1741 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     1801 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     1861 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     1921 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     1981 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     2041 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     2101 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     2161 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     2221 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     2281 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     2341 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     2401 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     2461 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     2521 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     2581 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     2641 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     2701 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     2761 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     2821 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     2881 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     2941 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     3001 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     3061 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     3121 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     3181 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     3241 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     3301 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     3361 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     3421 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     3481 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     3541 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     3601 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     3661 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     3721 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
     3781 acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc
     3841 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
     3901 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
     3961 tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg
     4021 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
     4081 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     4141 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     4201 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     4261 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     4321 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
     4381 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
     4441 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     4501 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
     4561 taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta gagaacccac
     4621 tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagacccaag cttggtaccg
     4681 agctcggatc caaagaattc ggcacgaggg ggaggagcgc ggcggtagcg gtgaattttg
     4741 aggggtgggt cgggggcgcg cactcgccgc ccctggtgct gccggctccc ggagccgtgg
     4801 cgtgtcccgg ctgtcgccgc tcggctgtcg cgagccgccg cgggcagagt cccgggcggg
     4861 ggagggagga agccggagcc tcaggcacgt gaaagaaaag cacaagaaga aacttttaca
     4921 ggcgttgttg attggactag ggcaacgatt cccaaaatca ccagcaagag ttctgatggt
     4981 cagtcacaca gaagacggcc ttgcgtctgt gggtgttgga gactccattc taaagatata
     5041 aaaagtgaaa gaggagaagt acaaatgtct accacaagac gaaaacataa tgtgttatgg
     5101 tgtttaccgt aagctgtagt aaaatgagct caattgttga cagagatgac agtagtattt
     5161 ttgatggatt ggtggaagaa gatgacaagg acaaagcaaa aagagtatct agaaacaaat
     5221 cagaaaagaa acgtagagat cagttcaatg tcctcattaa ggagctgggg tctatgcttc
     5281 ctggtaacgc gagaaagatg gacaagtcta ctgttctaca gaagagcatt gattttttgc
     5341 gcaaacataa agagaccact gcacagtcag atgctagtga gattcgacag gactggaaac
     5401 ccacattcct tagtaatgaa gagtttacac agttaatgtt agaggctctt gatggttttt
     5461 ttttagcgat catgacagat ggaagtataa tatatgtatc tgagagtgta acttcgttac
     5521 ttgaacattt accatctgat cttgtggatc aaagtatatt taattttatc ccagagggag
     5581 aacattcaga ggtttataag atactctcta ctcatctgct ggaaagtgac tcattaaccc
     5641 ctgagtactt aaaatcaaaa aatcagttag aattctgttg tcacatgctt cgaggaacaa
     5701 tagacccaaa ggagccatcc acctatgaat atgtgagatt tataggaaat tttaaatctt
     5761 taaccagtgt atcaacttca acacacaatg gttttgaagg aactatacaa cgcacacata
     5821 ggccttctta tgaagataga gtttgttttg tagctactgt cagattagct acacctcagt
     5881 tcatcaagga aatgtgtact gttgaagaac caaatgaaga gtttacatct agacacagtt
     5941 tagaatggaa gtttctattt ttagatcaca gggcaccacc aataataggc tatttgccat
     6001 ttgaagtctt gggaacatca ggctatgatt actatcatgt ggatgaccta gaaaatctgg
     6061 caaaatgtca cgagcactta atgcaatatg gaaaaggcaa atcgtgttac tatagattcc
     6121 tgaccaaagg ccagcagtgg atatggcttc agactcatta ttatattact taccatcagt
     6181 ggaattcaag gccagagttc attgtttgta ctcacactgt agtaagttat gcagaagtta
     6241 gggctgaaag acggcgagaa cttggcattg aagagtctct tcctgagaca gctgctgaca
     6301 aaagccaaga ttctgggtct gacaatcgta tcaacacagt gagtctcaag gaagcactgg
     6361 aaaggtttga tcacagccca actccttctg cctcctctag aagctcacga aagtcatctc
     6421 acaccgcagt ctcagaccct tcctccacac cgacaaagat ccctactgat actagcactc
     6481 ctcccagaca gcatttgcca gctcatgaaa agatgacaca gcggaggtcg tccttcagca
     6541 gtcagtccat aaactcccag tcagttggtc catcattaac acagccagcg atgtctcaag
     6601 ctgcaaattt accaattcca caaggcatgt cacagatgtt tttgcagcaa tcaaaccctg
     6661 gattgaattt tggttctgtt caactttcct ctggaaattc taatatccag caactcacac
     6721 ctgtaaatat gcaaggccag gttgtccctg ctaaccaggt tcagagtgga catatcagca
     6781 caggccagca catgatacag caacagactt tacaaagtac atcaactcag cagagtcaac
     6841 agagtgtaat gagtggacac agtcagcaga cgtctcttcc aagtcagaca ccgagcactc
     6901 tcacagcccc actgtacaat acgatggtga tttcccagcc tgcagctggg agcatggtcc
     6961 agattccatc cagtatgcca cagaacagta cccagagtgc tacagtcact acgttcactc
     7021 aggacagaca gataagattt tctcaaggtc agcaacttgt gaccaaatta gtgactgctc
     7081 ctgtagcttg tggggccgtc atggtaccaa gtaccatgct tatgggtcag gtggtgactg
     7141 cctatcctac cttcgccaca caacagcagc aggcacagac attatcggta acacaacagc
     7201 agcagcagca gcagcagcag ccaccacagc aacagcaaca acaacagcag agttcccagg
     7261 aacagcagct tccttcagtt cagcagccag ctcaggccca gctgggccag ccaccacagc
     7321 agttcttaca gacatctagg ttgctccacg ggaatccttc gacacagctc atcctctctg
     7381 ctgcctttcc actacaacag agcactttcc ctccttcgca ccaccagcaa caccagcctc
     7441 agcagcaaca gcagcttcct cggcacagga ctgacagcct gactgaccct tccaaggtcc
     7501 agccacagta gcacacacac ttcctctctg acatgcgaga ggaaggggat ggccagaaag
     7561 aatcgctcag ttggcatgcg gtcagaagtt gaacagtttc acgagggtgg tcttgagtgt
     7621 tcagtccctt gatgagacgg tagggaagtg ctgcccagtg cttcagatgt ccattaaata
     7681 ccagccagtg ggaaatggtc atagggacac agccaattct gacagtttct ttgcccaggt
     7741 attttttgat agaaagagta tattgccaaa tgctaacaag ctcagctatc aaccagatct
     7801 ttactgaatc cgaagagcac taacagtgtt ggtagcttta gtgggtctgt gcctgcatca
     7861 aatattacag agggcacacc actgccaggg gtttgcttag aatgccatga agatagtcca
     7921 gtagttaata gtccccaccc caaactcctc tccctgttca gacaatgatg gaaccgtgat
     7981 gactttgaga atgttgtgca ggtttgaatt cctgcagccc gggggatcca ctagtaacgg
     8041 ccgccagtgt gctggaattc tgcagatatc catcacactg gcggccgctc gagcatgcat
     8101 ctagagggcc ctattctata gtgtcaccta aatgctagag ctcgctgatc agcctcgact
     8161 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     8221 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     8281 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     8341 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     8401 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     8461 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     8521 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     8581 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     8641 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     8701 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     8761 cctatctcgg tctattcttt tga
//
