LOCUS       sequence-301930-        6561 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 6561)
  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 6561)
  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 CAT
FEATURES             Location/Qualifiers
     source          1..6561
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      684..693
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1785..1811
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(1880..1898)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1881..1898)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1918..1935)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1924..2148
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2194..2622
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2281..2300)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2491..2512
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2631..2651)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2636..2965
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2816..2951
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3032..3826
                     /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(3086..3105)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3696..3715
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4000..4121
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4037..4056)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4091..4110
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4170..4186)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4170..4186)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4183..4205)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4194..4210
                     /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(4218..4224,4225..4242,4243..4248))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4263..4284
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4401..4418)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4572..5160)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4652..4671)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5331..6122,6123..6191))
                     /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     5954..5973
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6192..6296)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6371..6390)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ggtaccgagc tcggatcctc gaggccacca tggaattctt ctttggtgcc agcccaggac
      721 gcctgcatat ctctgacttc agcttcctca tggttctagg gaaaggcagt tttgggaagg
      781 taatgctggc agagcgcaga ggctccgacg aactctatgc catcaagata ctgaagaaag
      841 acgtcatcgt ccaggatgac gatgtagact gcaccctcgt agagaagcgt gtcctggcat
      901 tgggaggccg aggtcctgga ggccggccac actttctcac gcagcttcac tccacctttc
      961 agactccgga ccgcctgtat tttgtgatgg agtatgtcac tgggggcgat ttaatgtacc
     1021 acatccagca actgggcaag tttaaggagc ctcatgcagc attctacgct gcggaaatcg
     1081 ccataggcct cttcttcctt cacaaccagg gcatcatcta cagggacctc aagttggata
     1141 atgtgatgct ggatgctgaa ggacacatca agatcacaga ctttggcatg tgtaaagaga
     1201 atgtcttccc tgggtccaca acccgcacct tctgtggcac cccagactac atagcacctg
     1261 agatcattgc ctatcagccc tacgggaagt ctgtcgactg gtggtctttt ggggtcctgc
     1321 tgtatgagat gttggcagga cagccaccct ttgatgggga agatgaggag gagttgtttc
     1381 aagccatcat ggaacaaact gtcacctatc ccaagtcact ttcccgggaa gctgtggcca
     1441 tctgcaaagg gttcctgacc aagcacccag gaaaacgcct gggctcaggg ccagatgggg
     1501 aacccaccat ccgggctcat ggctttttcc gttggatcga ttgggagagg ttggagagac
     1561 tggaaattgc acctcctttc agaccacgtc cgtgtggccg cagtggcgaa aactttgaca
     1621 agttcttcac gcgggcagcg ccagcactga ccccgccaga ccgcttggtt ctagccagca
     1681 tcgaccaagc tgatttccag ggctttactt atgtgaaccc ggacttcgtg cacccagatg
     1741 cccgcagccc cacaagccct gtgcccgtgc ctgtcatgga attctatcct tacgacgtgc
     1801 ctgactacgc ctaaggatcc aattctgcag atatccatca cactggcggc cgctcgagca
     1861 tgcatctaga gggccctatt ctatagtgtc acctaaatgc tagagctcgc tgatcagcct
     1921 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     1981 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     2041 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     2101 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     2161 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     2221 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     2281 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     2341 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     2401 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     2461 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     2521 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     2581 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     2641 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     2701 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     2761 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     2821 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     2881 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     2941 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     3001 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     3061 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     3121 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     3181 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     3241 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     3301 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     3361 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     3421 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     3481 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     3541 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     3601 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     3661 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     3721 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     3781 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     3841 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     3901 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     3961 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     4021 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     4081 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     4141 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     4201 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     4261 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     4321 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4381 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4441 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4501 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4561 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4621 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4681 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4741 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4801 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4861 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4921 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4981 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     5041 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5101 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5161 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5221 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     5281 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     5341 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     5401 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     5461 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     5521 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     5581 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     5641 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     5701 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     5761 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     5821 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     5881 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     5941 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     6001 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     6061 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     6121 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     6181 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     6241 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     6301 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
     6361 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
     6421 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
     6481 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
     6541 tacgggccag atatacgcgt t
//
