LOCUS       sequence-298544-        6519 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 6519)
  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 6519)
  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 II CAT
FEATURES             Location/Qualifiers
     source          1..6519
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        139..518
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        519..722
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     673..693
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     767..786
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        767..785
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      827..836
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             839..865
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(1976..1994)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1977..1994)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2014..2031)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2020..2244
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2290..2718
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2377..2396)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2587..2608
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2727..2747)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2732..3061
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2912..3047
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2974..2993
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3128..3922
                     /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(3182..3201)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3792..3811
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4096..4217
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4133..4152)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4187..4206
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4266..4282)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4266..4282)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4279..4301)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4290..4306
                     /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(4314..4320,4321..4338,4339..4344))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4359..4380
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4497..4514)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4668..5256)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4748..4767)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5427..6218,6219..6287))
                     /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     6050..6069
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6288..6392)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6467..6486)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
       61 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac
      121 gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
      181 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
      241 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
      301 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      361 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
      421 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
      481 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
      541 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
      601 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
      661 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
      721 ctctctggct aactagagaa cccactgctt actggcttat cgaaattaat acgactcact
      781 atagggagac ccaagcttgg taccgagctc ggatcgaatt ctcgaggcca ccatggctta
      841 tccttacgac gtgcctgact acgccggatc cgacaacaat ggcaacaggg accggatgaa
      901 actgaccgat tttaacttcc tgatggtgct ggggaaaggc agctttggca aggtcatgct
      961 ctcagagcgg aagggtacag atgaactcta tgccgtgaag atcctgaaga aggacgtggt
     1021 gatccaagat gacgatgtgg agtgcaccat ggtggagaag cgggtgctgg ccctgcctgg
     1081 gaagcccccg ttcctgactc agctccattc ctgcttccag accatggacc gcctgtactt
     1141 tgtgatggag tatgtgaacg ggggtgacct catgtaccac atccaacaag ttggccgttt
     1201 caaggagccc catgctgtat tttacgccgc agagattgcc atcggtctgt tcttcttgca
     1261 gagcaagggc attatttacc gcgacctgaa acttgacaac gtgatgctgg attctgaggg
     1321 gcacatcaaa atcgctgact ttggcatgtg taaggagaac atctgggatg gggtgacaac
     1381 caagacattc tgtggcactc cggactacat tgctccagag atcattgctt atcagcccta
     1441 cgggaagtct gtggactggt gggcatttgg agtcctgctg tatgaaatgt tggctggcca
     1501 ggcaccgttt gaaggagagg atgaggatga actcttccag tcaatcatgg aacacaatgt
     1561 ggcgtatccc aagtccatgt ccaaggaagc cgtggcaatc tgcaaagggc taatgaccaa
     1621 acacccaggc aagcgcctgg gttgtgggcc tgaaggggaa cgagacatta aggagcatgc
     1681 gtttttccgg tatattgact gggagaaact cgaacgcaag gagattcagc caccttataa
     1741 accaaaagct tgtgggcgaa acgctgaaaa cttcgaccgg tttttcaccc gccatccacc
     1801 agtcctaaca cctcctgacc aggaagtcat caggaatatt gaccaatcag aattcgaagg
     1861 attttccttt gttaactctg aatttttaaa acctgaagtc aagagcggat cctaagaatt
     1921 ctgcagatat ccatcacact ggcggccgct cgagcatgca tctagagggc cctattctat
     1981 agtgtcacct aaatgctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc
     2041 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     2101 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     2161 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     2221 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     2281 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     2341 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     2401 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     2461 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     2521 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     2581 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     2641 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     2701 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     2761 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     2821 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     2881 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     2941 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3001 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3061 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc
     3121 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     3181 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     3241 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     3301 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     3361 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     3421 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     3481 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     3541 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     3601 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     3661 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     3721 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     3781 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     3841 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     3901 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     3961 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     4021 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     4081 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     4141 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     4201 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     4261 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     4321 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     4381 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     4441 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     4501 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4561 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4621 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4681 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4741 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4801 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4861 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4921 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4981 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5041 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5101 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5161 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5221 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5281 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5341 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5401 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     5461 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     5521 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     5581 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     5641 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     5701 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     5761 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     5821 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     5881 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     5941 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6001 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6061 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6121 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6181 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     6241 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     6301 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     6361 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     6421 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
     6481 atgccgcata gttaagccag tatctgctcc ctgcttgtg
//
