LOCUS       sequence-138838-        7494 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 7494)
  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 7494)
  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 alpha DN
FEATURES             Location/Qualifiers
     source          1..7494
                     /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      2855..2864
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4889..4915
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(4984..5002)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4985..5002)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5022..5039)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5028..5252
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5298..5726
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5385..5404)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5595..5616
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5735..5755)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5740..6069
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5920..6055
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5982..6001
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6136..6930
                     /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(6190..6209)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6800..6819
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7104..7225
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7141..7160)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7195..7214
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7274..7290)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7274..7290)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7287..7309)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7298..7314
                     /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(7322..7328,7329..7346,7347..7352))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7367..7388
                     /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 ctcggatcct cgaggccacc atggaattcg ctgacgtttt
     2881 cccgggcaac gactccacgg cgtctcagga cgtggccaac cgcttcgccc gcaaaggggc
     2941 gctgaggcag aagaacgtgc acgaggtgaa ggaccacaaa ttcatcgcgc gcttcttcaa
     3001 gcagcccacc ttctgcagcc actgcaccga cttcatctgg gggtttggga aacaaggctt
     3061 ccagtgccaa gtttgctgtt ttgtggtcca caagaggtgc catgaatttg ttactttttc
     3121 ttgtccgggt gcggataagg gacccgacac tgatgacccc aggagcaagc acaagttcaa
     3181 aatccacact tacggaagcc ccaccttctg cgatcactgt gggtcactgc tctatggact
     3241 tatccatcaa gggatgaaat gtgacacctg cgatatgaac gttcacaagc aatgcgtcat
     3301 caatgtcccc agcctctgcg gaatggatca cactgagaag agggggcgga tttacctaaa
     3361 ggctgaggtt gctgatgaaa agctccatgt cacagtacga gatgcaaaaa atctaatccc
     3421 tatggatcca aacgggcttt cagatcctta tgtgaagctg aaacttattc ctgatcccaa
     3481 gaatgaaagc aagcaaaaaa ccaaaaccat ccgctccaca ctaaatccgc agtggaatga
     3541 gtcctttaca ttcaaattga aaccttcaga caaagaccga cgactgtctg tagaaatctg
     3601 ggactgggat cgaacaacaa ggaatgactt catgggatcc ctttcctttg gagtttcgga
     3661 gctgatgaag atgccggcca gtggatggta caagttgctt aaccaagaag aaggtgagta
     3721 ctacaacgta cccattccgg aaggggacga ggaaggaaac atggaactca ggcagaaatt
     3781 cgagaaagcc aaacttggcc ctgctggcaa caaagtcatc agtccctctg aagacaggaa
     3841 acaaccttcc aacaaccttg accgagtgaa actcacggac ttcaatttcc tcatggtgtt
     3901 gggaaagggg agttttggaa aggtgatgct tgccgacagg aagggcacag aagaactgta
     3961 tgcaatcaga atcctgaaga aggatgtggt gattcaggat gatgacgtgg agtgcaccat
     4021 ggtagaaaag cgagtcttgg ccctgcttga caaacccccg ttcttgacgc agctgcactc
     4081 ctgcttccag acagtggatc ggctgtactt cgtcatggaa tatgtcaacg gtggggacct
     4141 catgtaccac attcagcaag taggaaaatt taaggaacca caagcagtat tctatgcggc
     4201 agagatttcc atcggattgt tctttcttca taaaagagga atcatttata gggatctgaa
     4261 gttagataac gtcatgttgg attcagaagg acatatcaaa attgctgact ttgggatgtg
     4321 caaggaacac atgatggatg gagtcacgac caggaccttc tgtgggactc cagattatat
     4381 cgccccagag ataatcgctt atcagccgta tggaaaatct gtggactggt gggcctatgg
     4441 cgtcctgttg tatgaaatgc ttgccgggca gcctccattt gatggtgaag atgaagacga
     4501 gctatttcag tctatcatgg agcacaacgt ttcctatcca aaatccttgt ccaaggaggc
     4561 tgtttctatc tgcaaaggac tgatgaccaa acacccagcc aagcggctgg gctgtgggcc
     4621 tgagggggag agggacgtga gagagcatgc cttcttccgg aggatcgact gggaaaaact
     4681 ggagaacagg gagatccagc caccattcaa gcccaaagtg tgtggcaaag gagcagagaa
     4741 ctttgacaag ttcttcacac gaggacagcc cgtcttaaca ccacctgatc agctggttat
     4801 tgctaacata gaccagtctg attttgaagg gttctcgtat gtcaaccccc agtttgtgca
     4861 ccccatctta cagagtgcag tagaattcta tccttacgac gtgcctgact acgcctaagg
     4921 atccaattct gcagatatcc atcacactgg cggccgctcg agcatgcatc tagagggccc
     4981 tattctatag tgtcacctaa atgctagagc tcgctgatca gcctcgactg tgccttctag
     5041 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     5101 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     5161 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     5221 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     5281 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     5341 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     5401 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     5461 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     5521 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     5581 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     5641 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     5701 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     5761 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     5821 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     5881 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     5941 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     6001 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     6061 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     6121 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     6181 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     6241 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     6301 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     6361 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     6421 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     6481 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     6541 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     6601 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     6661 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     6721 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     6781 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     6841 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     6901 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     6961 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     7021 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     7081 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     7141 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     7201 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     7261 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     7321 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     7381 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     7441 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgct
//
