LOCUS       sequence-248149-        7491 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 7491)
  AUTHORS   Soh JW, Weinstein IB
  TITLE     Roles of specific isoforms of protein kinase C in the 
            transcriptional control of cyclin D1 and related genes.
  JOURNAL   J Biol Chem. 2003 Sep 5. 278(36):34709-16.
  PUBMED    12794082
REFERENCE   2  (bases 1 to 7491)
  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"; journalName: "J Biol 
            Chem. 2003 Sep 5. 278(36):34709-16."
COMMENT     Sequence Label: PKC beta I DN
FEATURES             Location/Qualifiers
     source          1..7491
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        48..427
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        428..631
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     582..602
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     676..695
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        676..694
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      736..745
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2767..2793
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(2857..2875)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2858..2875)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2895..2912)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2901..3125
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3171..3599
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3258..3277)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3468..3489
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3608..3628)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3613..3942
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3793..3928
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3855..3874
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4009..4803
                     /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(4063..4082)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4673..4692
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4977..5098
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5014..5033)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5068..5087
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5147..5163)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5147..5163)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5160..5182)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5171..5187
                     /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(5195..5201,5202..5219,5220..5225))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5240..5261
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5378..5395)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5549..6137)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5629..5648)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6308..7099,7100..7168))
                     /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     6931..6950
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7169..7273)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7348..7367)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac attgattatt
       61 gactagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
      121 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
      181 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
      241 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
      301 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
      361 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
      421 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
      481 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
      541 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
      601 tgtacggtgg gaggtctata taagcagagc tctctggcta actagagaac ccactgctta
      661 ctggcttatc gaaattaata cgactcacta tagggagacc caagcttggt accgagctcg
      721 gatcgaattc tcgaggccac catgggatcc gctgacccgg ctgcggggcc gccgccgagc
      781 gagggcgagg agagcacggt gcgcttcgcc cgcaaaggcg ccctccggca gaagaacgtg
      841 cacgaggtga agaaccacaa attcaccgcc cgcttcttca agcagcccac cttctgcagc
      901 cactgcaccg acttcatttg gggcttcggg aagcagggat tccagtgtca agtctgctgc
      961 tttgttgtac acaagcgctg ccatgaattc gtcacgttct cctgccctgg tgcagacaag
     1021 ggcccggcct ctgatgaccc acggagcaaa cacaagttta agatccacac ctactccagc
     1081 cctaccttct gtgaccactg tggatcactg ctgtatgggc tcatccacca ggggatgaaa
     1141 tgcgacacct gtatgatgaa tgtccacaag cgctgcgtga tgaacgtccc cagcctctgt
     1201 ggcaccgacc acacagaacg ccgtggccgc atctacatcc aggcccacat cgacagggag
     1261 gtcctcatcg ttgttgtaag agatgctaaa aatctggtac ctatggaccc caacggcttg
     1321 tcagatccct acgtaaaact gaaactgatc cctgatccca aaagtgagag caagcagaag
     1381 accaagacta tcaaatgctc cctcaacccg gagtggaacg aaaccttcag atttcagctg
     1441 aaggaatcag acaaagacag aagactgtcc gtagagatct gggattggga cctgaccagc
     1501 aggaatgact tcatgggatc tctgtcgttt gggatttcag aactacagaa agccggagtg
     1561 gatggctggt tcaagttact aagccaggaa gaaggcgagt actttaatgt gccggtgccg
     1621 ccggaaggaa gcgagggcaa tgaagagctg cggcagaagt ttgagagagc caagattggc
     1681 caaggtacca aggctccaga agaaaagaca gcgaacacta tatccaaatt tgacaacaat
     1741 ggcaacaggg accggatgaa actgaccgat tttaacttcc tgatggtgct ggggaaaggc
     1801 agctttggca aggtcatgct ctcagagcgg aagggtacag atgaactcta tgccgtgaga
     1861 atcctgaaga aagatgtggt gatccaagat gacgatgtgg agtgcacaat ggtggagaag
     1921 agggtgctgg ccctgcctgg gaagccccca ttcctgactc agctccattc ctgcttccag
     1981 accatggacc gcctctactt tgtgatggag tatgtgaacg ggggcgacct catgtaccac
     2041 atccaacaag ttggccgttt caaggagccc catgctgtat tttacgctgc agagattgcc
     2101 atcggtcttt tcttcttgca gagcaagggc atcatttacc gtgacctgaa acttgacaac
     2161 gtgatgctgg attccgaggg gcacatcaaa atcgctgact ttggcatgtg taaagagaat
     2221 atctgggatg gggtgacaac caagacattc tgtggcactc cagactacat tgccccagag
     2281 atcattgctt atcagcccta cgggaagtct gtggactggt gggcgtttgg agtcctgctg
     2341 tatgaaatgt tggctggcca ggcacctttt gaaggggagg atgaggatga actcttccag
     2401 tcaatcatgg agcacaacgt ggcgtatccc aagtccatgt ctaaggaagc tgtggcaatc
     2461 tgcaaagggc taatgaccaa acacccaggc aagcgcctgg gttgtgggcc tgaaggggaa
     2521 cgagacatta aggagcatgc atttttccgg tatatcgact gggagaaact cgaacgcaag
     2581 gagattcagc caccttataa accaaaagct agagacaagc gagacacctc caacttcgac
     2641 aaagagttca ccaggcagcc tgtggaactg actcccactg acaaactctt catcatgaac
     2701 ttggaccaaa atgaatttgc tggcttctcg tatactaacc cagagtttgt cattaatgtg
     2761 ggatcctatc cttacgacgt gcctgactac gcctaagaat tctgcagata tccatcacac
     2821 tggcggccgc tcgagcatgc atctagaggg ccctattcta tagtgtcacc taaatgctag
     2881 agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     2941 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     3001 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     3061 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     3121 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     3181 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3241 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3301 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     3361 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     3421 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3481 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     3541 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     3601 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3661 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     3721 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3781 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3841 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     3901 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     3961 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     4021 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     4081 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     4141 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     4201 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     4261 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     4321 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     4381 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     4441 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     4501 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     4561 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     4621 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     4681 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     4741 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     4801 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     4861 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     4921 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     4981 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     5041 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     5101 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     5161 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     5221 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     5281 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     5341 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     5401 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     5461 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     5521 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5581 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5641 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5701 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5761 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5821 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5881 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5941 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     6001 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     6061 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     6121 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     6181 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6241 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     6301 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6361 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6421 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6481 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6541 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6601 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6661 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6721 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6781 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6841 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6901 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6961 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     7021 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     7081 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     7141 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     7201 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     7261 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     7321 tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     7381 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
     7441 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg t
//
