LOCUS       sequence-174275-        7500 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7500)
  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 7500)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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 delta DN
FEATURES             Location/Qualifiers
     source          1..7500
                     /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             2724..2750
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(2819..2837)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2820..2837)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2857..2874)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2863..3087
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3133..3561
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3220..3239)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3430..3451
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3570..3590)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3575..3904
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3755..3890
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3817..3836
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3971..4765
                     /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(4025..4044)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4635..4654
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4939..5060
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4976..4995)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5030..5049
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5109..5125)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5109..5125)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5122..5144)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5133..5149
                     /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(5157..5163,5164..5181,5182..5187))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5202..5223
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5340..5357)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5511..6099)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5591..5610)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6270..7061,7062..7130))
                     /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     6893..6912
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7131..7235)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7310..7329)
                     /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 tggaattcgc acccttcctg cgcatctcct
      721 tcaattccta tgagctgggc tccctgcaag ttgaggacga agcaagccag cctttctgtg
      781 ctgtgaagat gaaggaggca ctcagcacag agcgagggaa gacactggta cagaagaagc
      841 ccaccatgta tcctgagtgg aagacaacgt tcgacgccca catctatgaa ggccgtgtta
      901 tccagattgt gctgatgcgg gcagctgaag accccgtgtc tgaggtcacg gtgggcgtgt
      961 cagtactggc tgagcgctgc aagaagaaca acggcaaggc tgagttctgg ctggatctgc
     1021 agcctcaggc caaggtgctg atgtgtgtgc agtatttcct ggaggatggg gattgcaaac
     1081 agtctatgcg tagtgaggag gaggcaaagt ttccaaccat gaaccgtcgt ggagccatta
     1141 aacaggccaa gatccactac atcaagaacc acgagtttat cgccaccttc ttcgggcagc
     1201 ccaccttctg ttctgtgtgc aaagagtttg tctggggcct caacaagcaa ggctacaaat
     1261 gcaggcaatg caacgctgcc atccacaaga aatgcattga caagattatc ggccgctgca
     1321 ctggcactgc caccaatagc cgggacacca tcttccagaa agaacgcttc aacatcgaca
     1381 tgcctcaccg attcaaggtt tataactaca tgagccccac cttctgtgac cactgtggca
     1441 gtttgctctg gggactggtg aagcagggat taaagtgtga agattgtggc atgaatgtgc
     1501 accacaaatg ccgggagaag gtggccaacc tgtgtggtat caaccaaaag ctcttggctg
     1561 aggccttgaa ccaagtgacc cagagatctt cccggaagct ggacacaaca gagtctgtcg
     1621 gaatatacca gggatttgag aagaagccag aagtctctgg gagtgacatc ctagacaaca
     1681 acgggaccta tggcaagatc tgggagggga gcacccggtg cacccttgag aacttcacct
     1741 tccaaaaagt acttggcaaa ggcagctttg gcaaggtgct gctggcagag ctgaagggca
     1801 aagacaagta ctttgcaatc aggtgtctga agaaggacgt ggtgttgatt gacgatgatg
     1861 tagagtgtac catggtggag aagcgggtgc tggcgctcgc ctgggagagt cccttcctca
     1921 cccacctcat ctgtaccttc cagaccaagg accacctgtt cttcgtgatg gagtttctca
     1981 atgggggtga cctgatgttc cacattcagg acaaaggccg cttcgaactc taccgggcta
     2041 cgttttatgc agctgagatc atctgcggac tgcagtttct acacagcaaa ggcattattt
     2101 acagggacct caagctggac aatgtgatgc tagacaggga cggccacatc aagatcgctg
     2161 actttgggat gtgcaaagag aatatatttg gggagggccg ggccagcaca ttctgcggca
     2221 ctcctgacta catcgcccct gagatcctgc agggcctgaa gtactccttc tcggtggact
     2281 ggtggtcttt cggggtcctc ctgtacgaaa tgctcatcgg ccagtccccc ttccacggcg
     2341 acgatgagga cgagctcttc gagtccatcc gggtggacac accacactat ccccgttgga
     2401 tcaccaagga atccaaggac atcatggaga agctattcga gagggaccct gacaagaggc
     2461 tgggagtaac aggaaacatc aggattcacc cctttttcaa gactatcaac tggtccctcc
     2521 tggagaagcg gaaggtggag ccgcccttta agcccaaagt gaaatcccct tcagactaca
     2581 gcaactttga cccagagttc ctgaatgaga aacctcagct ttccttcagt gacaagaacc
     2641 tcatcgactc tatggaccag gaagccttcc atggcttctc ctttgtgaat cccaagtttg
     2701 agcaattcct ggacattgaa ttctatcctt acgacgtgcc tgactacgcc taaggatcca
     2761 attctgcaga tatccatcac actggcggcc gctcgagcat gcatctagag ggccctattc
     2821 tatagtgtca cctaaatgct agagctcgct gatcagcctc gactgtgcct tctagttgcc
     2881 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     2941 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     3001 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     3061 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     3121 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     3181 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     3241 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     3301 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     3361 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     3421 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     3481 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     3541 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     3601 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     3661 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3721 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     3781 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     3841 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     3901 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg
     3961 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     4021 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     4081 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     4141 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     4201 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     4261 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     4321 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     4381 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     4441 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     4501 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     4561 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     4621 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     4681 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     4741 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     4801 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     4861 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     4921 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     4981 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     5041 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     5101 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     5161 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     5221 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     5281 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     5341 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     5401 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     5461 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     5521 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5581 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     5641 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     5701 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5761 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5821 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5881 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5941 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     6001 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     6061 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     6121 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     6181 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     6241 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     6301 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     6361 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     6421 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     6481 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     6541 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     6601 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     6661 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     6721 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     6781 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     6841 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     6901 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     6961 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     7021 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     7081 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     7141 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     7201 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     7261 gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct
     7321 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
     7381 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
     7441 atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt
//
