LOCUS       sequence-242477-        7305 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 7305)
  AUTHORS   Ye R, Selby CP, Ozturk N, Annayev Y, Sancar A
  TITLE     Biochemical analysis of the canonical model for the mammalian 
            circadian clock.
  JOURNAL   J Biol Chem. 2011 Jul 22;286(29):25891-902. doi: 
            10.1074/jbc.M111.254680. Epub 2011 May 25.
  PUBMED    21613214
REFERENCE   2  (bases 1 to 7305)
  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: 
            "10.1074/jbc.M111.254680"; journalName: "J Biol Chem"; date: 
            "2011-07-22- 22"; volume: "286"; issue: "29"; pages: "25891-902"
COMMENT     Sequence Label: pBMPC3
FEATURES             Location/Qualifiers
     source          1..7305
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        119..498
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        499..702
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     653..673
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     747..766
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        747..765
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2706..2723
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        complement(2742..2760)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2743..2760)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2780..2797)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2786..3010
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3056..3484
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3143..3162)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3353..3374
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3493..3513)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3498..3827
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3678..3813
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3740..3759
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3894..4688
                     /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(3948..3967)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4558..4577
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4862..4983
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4899..4918)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4953..4972
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5032..5048)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5032..5048)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5045..5067)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5056..5072
                     /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(5080..5086,5087..5104,5105..5110))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5125..5146
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5263..5280)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5434..6022)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5514..5533)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6193..6984,6985..7053))
                     /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     6816..6835
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7054..7158)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7233..7252)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
       61 ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga
      121 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
      181 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
      241 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
      301 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
      361 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      421 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
      481 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
      541 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
      601 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
      661 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctctctggct aactagagaa
      721 cccactgctt actggcttat cgaaattaat acgactcact atagggagac ccaagcttgg
      781 taccgagctc ggatccacta gtaacggccg ccagtgtgct ggaattcatg gcggaccaga
      841 gaatggacat ttcctcaacc atcagcgact tcatgtctcc gggccccacc gacctactct
      901 ccggttccct gggcaccagt ggtgtggact gcaatcgcaa gaggaaaggc agtgccactg
      961 actaccaaga aagtatggac acagacaaag atgaccctca tggaaggtta gaatatgcag
     1021 aacaccaagg aaggatcaag aatgcaaggg aggcccacag tcagattgaa aagaggcgtc
     1081 gggacaaaat gaacagtttc attgatgaat tggcttcttt ggtaccaaca tgcaatgcaa
     1141 tgtccaggaa gttagataaa ctcaccgtgc taaggatggc tgttcagcac atgaaaactt
     1201 tgagaggtgc caccaaccca tacacagaag caaactacaa gccaacattt ctatcagatg
     1261 acgaactgaa acacctaatt ctcagggcag cagatggatt tttgtttgtc gtaggatgtg
     1321 accgagggaa gatcctcttt gtctccgagt ctgtcttcaa gatcctcaat tatagccaga
     1381 atgaccttat tggccagagc ttgtttgact acctgcatcc aaaagatatt gccaaagtta
     1441 aggaacagct atcttcctcg gacactgcgc cccgggagcg actcattgat gccaagactg
     1501 gacttccggt taaaacggat ataacccctg ggccctcccg gctatgctct ggagcccgcc
     1561 gctctttctt ctgtagaatg aagtgcaaca ggccttcagt aaaggtggaa gataaggact
     1621 tcgcctctac ctgttcaaag aaaaaagcag atcgaaaaag cttctgcaca atccacagca
     1681 caggctattt gaaaagctgg ccacccacga agatggggct ggacgaagac aatgagccag
     1741 acaacgaggg ctgcaacctc agctgcctcg ttgcaatcgg gcgcctgcac tcgcacatgg
     1801 ttccacaacc agcgaacggg gaaatacggg tgaaatctat ggagtacgtt tctcgacacg
     1861 caatagatgg gaaatttgtt tttgtagatc agagggcgac agctattttg gcgtatctac
     1921 cacaggaact tctaggtaca tcatgttatg agtattttca tcaagacgac ataggacacc
     1981 tcgcagaatg tcacaggcaa gttttacaga caagagaaaa gatcacaact aattgctata
     2041 agtttaagat caaagatggt tcttttatca cgctacgaag tcgatggttc agtttcatga
     2101 acccgtggac caaggaagtt gaatacattg tctcaaccaa cactgttgtt ttagccaatg
     2161 tcctggaagg cggggaccca accttcccgc agctaacagc acccccccac agcatggaca
     2221 gcatgctgcc ctctggagaa ggtggcccaa agaggactca tcccactgtt ccaggcattc
     2281 cagggggaac cagagccgga gcaggaaaaa taggtcgaat gattgccgag gaaatcatgg
     2341 aaatccacag gataagaggg tcatcgcctt ccagctgtgg ctccagcccg ctgaacatca
     2401 caagtacgcc tccccctgat gcctcttctc caggaggcaa gaagattcta aatggaggga
     2461 ctccagacat tccttccact ggactattac cagggcaggc tcaggagacc ccagggtatc
     2521 cctattctga tagttcttct attcttggtg agaaccccca cataggcatc gatatgatag
     2581 ataacgacca aggatcaagt agtcccagta acgatgaggc agcaatggct gtcatcatga
     2641 gcctcttgga agcagatgcg gggctgggtg gccccgttga ctttagtgac ttgccatggc
     2701 cgctgcatca ccatcaccat cactagtcta gagggcccta ttctatagtg tcacctaaat
     2761 gctagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     2821 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     2881 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     2941 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     3001 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     3061 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3121 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3181 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     3241 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     3301 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3361 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     3421 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3481 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3541 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3601 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3661 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3721 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     3781 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     3841 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     3901 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     3961 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     4021 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     4081 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     4141 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     4201 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     4261 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     4321 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     4381 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     4441 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     4501 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     4561 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     4621 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     4681 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     4741 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     4801 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     4861 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     4921 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     4981 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     5041 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     5101 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     5161 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     5221 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     5281 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5341 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5401 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5461 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5521 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5581 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5641 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5701 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5761 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5821 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     5881 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5941 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     6001 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6061 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6121 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6181 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6241 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6301 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6361 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     6421 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6481 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     6541 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     6601 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     6661 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     6721 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6781 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     6841 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6901 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6961 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     7021 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     7081 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7141 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     7201 agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     7261 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtag
//
