LOCUS       sequence-287543-        7569 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 7569)
  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 7569)
  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 gamma DN
FEATURES             Location/Qualifiers
     source          1..7569
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      918..927
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3027..3053
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(3122..3140)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3123..3140)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3160..3177)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3166..3390
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3436..3864
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3523..3542)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3733..3754
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3873..3893)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3878..4207
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4058..4193
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4274..5068
                     /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(4328..4347)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4938..4957
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5242..5363
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5279..5298)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5333..5352
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5412..5428)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5412..5428)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5425..5447)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5436..5452
                     /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(5460..5466,5467..5484,5485..5490))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5505..5526
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5643..5660)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5814..6402)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5894..5913)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6573..7364,7365..7433))
                     /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     7196..7215
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7434..7538)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat cctcgaggcc accatggaat tcgcgggtct gggccctggc ggaggcgact
      961 cagagggggg accccgaccc ctgttttgca gaaagggggc gctgaggcag aaggtggtcc
     1021 acgaggtgaa gagccacaag ttcaccgctc gtttcttcaa gcagccaacc ttctgcagtc
     1081 actgtaccga cttcatctgg ggcattggaa agcagggcct gcaatgtcaa gtctgtagct
     1141 ttgtggttca ccgccgatgc cacgaatttg tgaccttcga gtgtccaggc gctggaaagg
     1201 gcccccagac ggacgaccct cgcaacaagc acaagttccg tctgcacagc tacagcagtc
     1261 ccaccttctg cgaccactgt ggctccctcc tttacgggct ggtgcaccag ggcatgaaat
     1321 gttcctgttg cgagatgaat gtgcaccggc gctgtgtgcg cagcgtgccc tccctttgcg
     1381 gtgtggacca cacagagcgc cgtggacgtc tgcaactgga aatccgggct cctacgtcgg
     1441 atgagatcca tattactgtt ggcgaggccc ggaacctcat tcctatggac cccaatggcc
     1501 tgtctgatcc ctacgtgaaa ctgaagctga tcccggaccc tcggaacctg acaaaacaga
     1561 agacaaagac cgtgaaagcc acactgaatc ccgtgtggaa cgagaccttc gtgttcaacc
     1621 tgaagccagg ggatgtagag cgccggctca gtgtggaggt gtgggattgg gataggacat
     1681 cccgaaatga cttcatgggt gccatgtcct ttggtgtctc agagctactc aaggcccctg
     1741 tggatggatg gtacaagtta ctgaaccagg aggagggcga atattacaat gtaccggtgg
     1801 ccgatgctga caactgcagc ctcctccaga agtttgaggc ctgcaattac cccttggaat
     1861 tgtatgagag agtgcggatg ggcccctctt cctcccccat cccttctcca tcccccagtc
     1921 ctacggactc caagcgatgc ttctttggtg ccagcccagg acgcctgcat atctctgact
     1981 tcagcttcct catggttcta gggaaaggca gttttgggaa ggtaatgctg gcagagcgca
     2041 gaggctccga cgaactctat gccatcagga tactgaagaa agacgtcatc gtccaggatg
     2101 acgatgtaga ctgcaccctc gtagagaagc gtgtcctggc attgggaggc cgaggtcctg
     2161 gaggccggcc acactttctc acgcagcttc actccacctt tcagactccg gaccgcctgt
     2221 attttgtgat ggagtatgtc actgggggcg atttaatgta ccacatccag caactgggca
     2281 agtttaagga gcctcatgca gcattctacg ctgcggaaat cgccataggc ctcttcttcc
     2341 ttcacaacca gggcatcatc tacagggacc tcaagttgga taatgtgatg ctggatgctg
     2401 aaggacacat caagatcaca gactttggca tgtgtaaaga gaatgtcttc cctgggtcca
     2461 caacccgcac cttctgtggc accccagact acatagcacc tgagatcatt gcctatcagc
     2521 cctacgggaa gtctgtcgac tggtggtctt ttggggtcct gctgtatgag atgttggcag
     2581 gacagccacc ctttgatggg gaagatgagg aggagttgtt tcaagccatc atggaacaaa
     2641 ctgtcaccta tcccaagtca ctttcccggg aagctgtggc catctgcaaa gggttcctga
     2701 ccaagcaccc aggaaaacgc ctgggctcag ggccagatgg ggaacccacc atccgggctc
     2761 atggcttttt ccgttggatc gattgggaga ggttggagag actggaaatt gcacctcctt
     2821 tcagaccacg tccgtgtggc cgcagtggcg aaaactttga caagttcttc acgcgggcag
     2881 cgccagcact gaccccgcca gaccgcttgg ttctagccag catcgaccaa gctgatttcc
     2941 agggctttac ttatgtgaac ccggacttcg tgcacccaga tgcccgcagc cccacaagcc
     3001 ctgtgcccgt gcctgtcatg gaattctatc cttacgacgt gcctgactac gcctaaggat
     3061 ccaattctgc agatatccat cacactggcg gccgctcgag catgcatcta gagggcccta
     3121 ttctatagtg tcacctaaat gctagagctc gctgatcagc ctcgactgtg ccttctagtt
     3181 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     3241 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     3301 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     3361 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     3421 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     3481 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     3541 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     3601 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     3661 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     3721 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     3781 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     3841 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     3901 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3961 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4021 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     4081 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     4141 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     4201 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg
     4261 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     4321 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     4381 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     4441 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     4501 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     4561 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     4621 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     4681 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     4741 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     4801 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     4861 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     4921 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     4981 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     5041 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     5101 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     5161 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     5221 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     5281 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     5341 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     5401 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     5461 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     5521 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     5581 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     5641 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     5701 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     5761 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     5821 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5881 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5941 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     6001 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     6061 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     6121 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     6181 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     6241 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     6301 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     6361 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     6421 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     6481 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     6541 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     6601 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     6661 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     6721 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     6781 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     6841 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     6901 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     6961 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     7021 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     7081 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     7141 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     7201 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     7261 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     7321 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     7381 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     7441 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     7501 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     7561 cctgacgtc
//
