LOCUS       sequence-140177-        7500 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 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 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 delta WT
FEATURES             Location/Qualifiers
     source          1..7500
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    270..391
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(281..300)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     335..354
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(565..1359)
                     /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(676..695)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1286..1305
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1426..1755)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1440..1575
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1494..1513)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     1740..1760
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(1769..2197)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1879..1900)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2091..2110
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    2243..2467
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2456..2473
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        2493..2511
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2493..2510
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             complement(2580..2606)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      4637..4646
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(4682..4701)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4683..4701)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4746..4949)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4775..4795)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4950..5329
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     5501..5520
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5595..5699
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5700..5768,5769..6560)
                     /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     complement(5918..5937)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6731..7319
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7220..7239
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7473..7490
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctct
      241 agctagaggt cgacggtata cagacatgat aagatacatt gatgagtttg gacaaaccac
      301 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
      361 tgtaaccatt ataagctgca ataaacaagt tggggtgggc gaagaactcc agcatgagat
      421 ccccgcgctg gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt
      481 catagaaggc ggcggtggaa tcgaaatctc gtgatggcag gttgggcgtc gcttggtcgg
      541 tcatttcgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat agaaggcgat
      601 gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag cccattcgcc
      661 gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc ggtccgccac
      721 acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca tgatattcgg
      781 caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcatgc gcgccttgag
      841 cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat catcctgatc
      901 gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc
      961 gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag ccatgatgga
     1021 tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca cttcgcccaa
     1081 tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc
     1141 cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc agttcattca gggcaccgga
     1201 caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga acacggcggc
     1261 atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct ccacccaagc
     1321 ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt
     1381 ctcttgatca gatccgaaaa tggatataca agctcccggg agctttttgc aaaagcctag
     1441 gcctccaaaa aagcctcctc actacttctg gaatagctca gaggcagagg cggcctcggc
     1501 ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg
     1561 agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact aattgagatg
     1621 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc tggttgctga
     1681 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
     1741 ccctaactga cacacattcc acagaattaa ttcgcgttaa atttttgtta aatcagctca
     1801 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     1861 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     1921 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     1981 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     2041 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     2101 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     2161 acacccgccg cgcttaatgc gccgctacag ggcgcgtggg gataccccct agagccccag
     2221 ctggttcttt ccgcctcaga agccatagag cccaccgcat ccccagcatg cctgctattg
     2281 tcttcccaat cctccccctt gctgtcctgc cccaccccac cccccagaat agaatgacac
     2341 ctactcagac aatgcgatgc aatttcctca ttttattagg aaaggacagt gggagtggca
     2401 ccttccaggg tcaaggaagg cacgggggag gggcaaacaa cagatggctg gcaactagaa
     2461 ggcacagtcg aggctgatca gcgagctcta gcatttaggt gacactatag aatagggccc
     2521 tctagatgca tgctcgagcg gccgccagtg tgatggatat ctgcagaatt ggatccttag
     2581 gcgtagtcag gcacgtcgta aggatagaat tcaatgtcca ggaattgctc aaacttggga
     2641 ttcacaaagg agaagccatg gaaggcttcc tggtccatag agtcgatgag gttcttgtca
     2701 ctgaaggaaa gctgaggttt ctcattcagg aactctgggt caaagttgct gtagtctgaa
     2761 ggggatttca ctttgggctt aaagggcggc tccaccttcc gcttctccag gagggaccag
     2821 ttgatagtct tgaaaaaggg gtgaatcctg atgtttcctg ttactcccag cctcttgtca
     2881 gggtccctct cgaatagctt ctccatgatg tccttggatt ccttggtgat ccaacgggga
     2941 tagtgtggtg tgtccacccg gatggactcg aagagctcgt cctcatcgtc gccgtggaag
     3001 ggggactggc cgatgagcat ttcgtacagg aggaccccga aagaccacca gtccaccgag
     3061 aaggagtact tcaggccctg caggatctca ggggcgatgt agtcaggagt gccgcagaat
     3121 gtgctggccc ggccctcccc aaatatattc tctttgcaca tcccaaagtc agcgatcttg
     3181 atgtggccgt ccctgtctag catcacattg tccagcttga ggtccctgta aataatgcct
     3241 ttgctgtgta gaaactgcag tccgcagatg atctcagctg cataaaacgt agcccggtag
     3301 agttcgaagc ggcctttgtc ctgaatgtgg aacatcaggt cacccccatt gagaaactcc
     3361 atcacgaaga acaggtggtc cttggtctgg aaggtacaga tgaggtgggt gaggaaggga
     3421 ctctcccagg cgagcgccag cacccgcttc tccaccatgg tacactctac atcatcgtca
     3481 atcaacacca cgtccttctt cagacacttg attgcaaagt acttgtcttt gcccttcagc
     3541 tctgccagca gcaccttgcc aaagctgcct ttgccaagta ctttttggaa ggtgaagttc
     3601 tcaagggtgc accgggtgct cccctcccag atcttgccat aggtcccgtt gttgtctagg
     3661 atgtcactcc cagagacttc tggcttcttc tcaaatccct ggtatattcc gacagactct
     3721 gttgtgtcca gcttccggga agatctctgg gtcacttggt tcaaggcctc agccaagagc
     3781 ttttggttga taccacacag gttggccacc ttctcccggc atttgtggtg cacattcatg
     3841 ccacaatctt cacactttaa tccctgcttc accagtcccc agagcaaact gccacagtgg
     3901 tcacagaagg tggggctcat gtagttataa accttgaatc ggtgaggcat gtcgatgttg
     3961 aagcgttctt tctggaagat ggtgtcccgg ctattggtgg cagtgccagt gcagcggccg
     4021 ataatcttgt caatgcattt cttgtggatg gcagcgttgc attgcctgca tttgtagcct
     4081 tgcttgttga ggccccagac aaactctttg cacacagaac agaaggtggg ctgcccgaag
     4141 aaggtggcga taaactcgtg gttcttgatg tagtggatct tggcctgttt aatggctcca
     4201 cgacggttca tggttggaaa ctttgcctcc tcctcactac gcatagactg tttgcaatcc
     4261 ccatcctcca ggaaatactg cacacacatc agcaccttgg cctgaggctg cagatccagc
     4321 cagaactcag ccttgccgtt gttcttcttg cagcgctcag ccagtactga cacgcccacc
     4381 gtgacctcag acacggggtc ttcagctgcc cgcatcagca caatctggat aacacggcct
     4441 tcatagatgt gggcgtcgaa cgttgtcttc cactcaggat acatggtggg cttcttctgt
     4501 accagtgtct tccctcgctc tgtgctgagt gcctccttca tcttcacagc acagaaaggc
     4561 tggcttgctt cgtcctcaac ttgcagggag cccagctcat aggaattgaa ggagatgcgc
     4621 aggaagggtg cgaattccat ggtggcctcg aggatccgag ctcggtacca agcttgggtc
     4681 tccctatagt gagtcgtatt aatttcgata agccagtaag cagtgggttc tctagttagc
     4741 cagagagctc tgcttatata gacctcccac cgtacacgcc taccgcccat ttgcgtcaat
     4801 ggggcggagt tgttacgaca ttttggaaag tcccgttgat tttggtgcca aaacaaactc
     4861 ccattgacgt caatggggtg gagacttgga aatccccgtg agtcaaaccg ctatccacgc
     4921 ccattgatgt actgccaaaa ccgcatcacc atggtaatag cgatgactaa tacgtagatg
     4981 tactgccaag taggaaagtc ccataaggtc atgtactggg cataatgcca ggcgggccat
     5041 ttaccgtcat tgacgtcaat agggggcgta cttggcatat gatacacttg atgtactgcc
     5101 aagtgggcag tttaccgtaa atactccacc cattgacgtc aatggaaagt ccctattggc
     5161 gttactatgg gaacatacgt cattattgac gtcaatgggc gggggtcgtt gggcggtcag
     5221 ccaggcgggc catttaccgt aagttatgta acgcggaact ccatatatgg gctatgaact
     5281 aatgaccccg taattgatta ctattaataa ctagtcaata atcaatgtca acgcgtatat
     5341 ctggcccgta catcgcgaag cagcgcaaaa cgcctaaccc taagcagatt cttcatgcaa
     5401 ttgtcggtca agccttgcct tgttgtagct taaattttgc tcgcgcacta ctcagcgacc
     5461 tccaacacac aagcagggag cagatactgg cttaactatg cggcatcaga gcagattgta
     5521 ctgagagtgc accatagggg atcgggagat ctcccgatcc gtcgacgtca ggtggcactt
     5581 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     5641 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     5701 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     5761 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     5821 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     5881 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     5941 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     6001 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     6061 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     6121 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     6181 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     6241 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     6301 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     6361 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     6421 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     6481 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     6541 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     6601 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     6661 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     6721 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     6781 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     6841 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     6901 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     6961 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     7021 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     7081 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     7141 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     7201 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     7261 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     7321 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     7381 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     7441 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
//
