LOCUS       sequence-134985-        7425 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 7425)
  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 7425)
  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 DN
FEATURES             Location/Qualifiers
     source          1..7425
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(11..28)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(941..1732,1733..1801))
                     /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     1564..1583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1802..1906)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1981..2000)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2172..2551
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2552..2755
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2706..2726
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2800..2819
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2800..2818
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2860..2869
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4897..4923
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(4987..5005)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4988..5005)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5025..5042)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5031..5255
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5301..5729
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5388..5407)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5598..5619
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5738..5758)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5782..5992
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        5804..6000
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5851..5986
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5913..5932
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6067..6861
                     /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(6121..6140)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6731..6750
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7035..7156
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7072..7091)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7126..7145
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7205..7221)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7205..7221)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7218..7240)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7229..7245
                     /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(7253..7259,7260..7277,7278..7283))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7298..7319
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag
     1981 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga
     2041 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa
     2101 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag
     2161 atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
     2221 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     2281 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     2341 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     2401 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     2461 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     2521 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
     2581 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     2641 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     2701 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctctg
     2761 gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc actataggga
     2821 gacccaagct tggtaccgag ctcggatcga attctcgagg ccaccatggg atccgctgac
     2881 ccggctgcgg ggccgccgcc gagcgagggc gaggagagca cagtgcgctt cgcccgcaaa
     2941 ggcgccctcc ggcagaagaa cgtgcacgag gtgaagaacc acaaattcac cgcccgcttc
     3001 ttcaagcagc ccaccttctg cagccactgc accgacttca tctggggctt cgggaagcag
     3061 ggattccagt gtcaagtctg ctgctttgtt gtacacaagc gctgccatga gttcgtcacg
     3121 ttctcctgcc ccggtgcgga caagggcccg gcctctgatg acccccggag caaacacaag
     3181 tttaagatcc acacgtactc cagtcctacc ttctgtgacc actgtggatc gctgctgtat
     3241 ggacttattc accaggggat gaaatgcgac acctgtatga tgaacgtgca caagcgctgc
     3301 gtgatgaacg tccccagcct ctgcggcact gaccacacgg aacgccgcgg ccgcatctat
     3361 atccaggccc acatcgacag agaggttctc attgttgttg taagagatgc taaaaatctg
     3421 gtacctatgg accccaatgg cttgtcagat ccctacgtta aactgaaact gatccctgat
     3481 cccaaaagtg agagcaaaca gaagaccaag accatcaagt gttccctcaa tccagagtgg
     3541 aatgaaacct tcagatttca gctgaaggaa tcagacaaag acagaagact gtctgtagag
     3601 atctgggatt gggacctgac cagcaggaat gacttcatgg gatctctgtc atttgggatt
     3661 tcagaactac agaaagccgg tgtggatggc tggttcaagt tactaagcca ggaagagggt
     3721 gaatacttca atgtgcccgt gccgcccgaa ggaagcgagg gcaatgaaga gctgcggcag
     3781 aagtttgaga gagccaagat tggccaaggt accaaggctc cagaagaaaa gacagctaac
     3841 actatctcca aatttgacaa caatggcaac agggaccgga tgaaactgac cgattttaac
     3901 ttcctgatgg tgctggggaa aggcagcttt ggcaaggtca tgctctcaga gcggaagggt
     3961 acagatgaac tctatgccgt gagaatcctg aagaaggacg tggtgatcca agatgacgat
     4021 gtggagtgca ccatggtgga gaagcgggtg ctggccctgc ctgggaagcc cccgttcctg
     4081 actcagctcc attcctgctt ccagaccatg gaccgcctgt actttgtgat ggagtatgtg
     4141 aacgggggtg acctcatgta ccacatccaa caagttggcc gtttcaagga gccccatgct
     4201 gtattttacg ccgcagagat tgccatcggt ctgttcttct tgcagagcaa gggcattatt
     4261 taccgcgacc tgaaacttga caacgtgatg ctggattctg aggggcacat caaaatcgct
     4321 gactttggca tgtgtaagga gaacatctgg gatggggtga caaccaagac attctgtggc
     4381 actccggact acattgctcc agagatcatt gcttatcagc cctacgggaa gtctgtggac
     4441 tggtgggcat ttggagtcct gctgtatgaa atgttggctg gccaggcacc gtttgaagga
     4501 gaggatgagg atgaactctt ccagtcaatc atggaacaca atgtggcgta tcccaagtcc
     4561 atgtccaagg aagccgtggc aatctgcaaa gggctaatga ccaaacaccc aggcaagcgc
     4621 ctgggttgtg ggcctgaagg ggaacgagac attaaggagc atgcgttttt ccggtatatt
     4681 gactgggaga aactcgaacg caaggagatt cagccacctt ataaaccaaa agcttgtggg
     4741 cgaaacgctg aaaacttcga ccggtttttc acccgccatc caccagtcct aacacctcct
     4801 gaccaggaag tcatcaggaa tattgaccaa tcagaattcg aaggattttc ctttgttaac
     4861 tctgaatttt taaaacctga agtcaagagc ggatcctatc cttacgacgt gcctgactac
     4921 gcctaagaat tctgcagata tccatcacac tggcggccgc tcgagcatgc atctagaggg
     4981 ccctattcta tagtgtcacc taaatgctag agctcgctga tcagcctcga ctgtgccttc
     5041 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     5101 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     5161 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     5221 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     5281 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     5341 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     5401 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     5461 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     5521 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     5581 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     5641 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     5701 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     5761 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     5821 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     5881 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     5941 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     6001 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     6061 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     6121 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     6181 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     6241 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     6301 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     6361 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     6421 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     6481 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     6541 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     6601 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     6661 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     6721 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     6781 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     6841 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     6901 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     6961 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     7021 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     7081 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     7141 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     7201 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     7261 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     7321 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     7381 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgct
//
