LOCUS       sequence-354476-        6521 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 6521)
  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 6521)
  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 alpha CAT
FEATURES             Location/Qualifiers
     source          1..6521
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(80..100)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        85..414
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      265..400
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     327..346
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(535..554)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1144..1163
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1448..1569
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1485..1504)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1539..1558
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1618..1634)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1618..1634)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1631..1653)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1642..1658
                     /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(1666..1672,1673..1690,1691..1696))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1711..1732
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1849..1866)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2020..2608)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2100..2119)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2779..3570,3571..3639))
                     /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     3402..3421
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3640..3744)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3819..3838)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4010..4389
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4390..4593
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4544..4564
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4638..4657
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4638..4656
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4693..4702
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5755..5781
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(5850..5868)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5851..5868)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5888..5905)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5894..6118
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(6164..6521,1..71)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6251..6270)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6461..6482
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
       61 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
      121 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
      181 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
      241 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
      301 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
      361 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
      421 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
      481 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
      541 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
      601 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
      661 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
      721 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
      781 gatctcctgt catctcacct tgctctgccg agaaagtatc catcatggct gatgcaatgc
      841 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
      901 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
      961 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     1021 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     1081 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     1141 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     1201 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     1261 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     1321 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     1381 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     1441 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     1501 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     1561 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     1621 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     1681 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     1741 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     1801 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     1861 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     1921 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     1981 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     2041 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2101 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2161 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     2221 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     2281 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     2341 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     2401 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     2461 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     2521 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     2581 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     2641 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     2701 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     2761 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     2821 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     2881 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     2941 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     3001 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     3061 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     3121 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     3181 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     3241 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     3301 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     3361 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     3421 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     3481 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     3541 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     3601 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     3661 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     3721 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     3781 atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca
     3841 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
     3901 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
     3961 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
     4021 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
     4081 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
     4141 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
     4201 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
     4261 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
     4321 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     4381 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
     4441 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
     4501 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
     4561 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
     4621 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagcttg gtaccgagct
     4681 cggatcctcg aggccaccat ggaattccaa ccttccaaca accttgaccg agtgaaactc
     4741 acggacttca atttcctcat ggtgttggga aaggggagtt ttggaaaggt gatgcttgcc
     4801 gacaggaagg gcacagaaga actgtatgca atcaaaatcc tgaagaagga tgtggtgatt
     4861 caggatgatg acgtggagtg caccatggta gaaaagcgag tcttggccct gcttgacaaa
     4921 cccccgttct tgacgcagct gcactcctgc ttccagacag tggatcggct gtacttcgtc
     4981 atggaatatg tcaacggtgg ggacctcatg taccacattc agcaagtagg aaaatttaag
     5041 gaaccacaag cagtattcta tgcggcagag atttccatcg gattgttctt tcttcataaa
     5101 agaggaatca tttataggga tctgaagtta gataacgtca tgttggattc agaaggacat
     5161 atcaaaattg ctgactttgg gatgtgcaag gaacacatga tggatggagt cacgaccagg
     5221 accttctgtg ggactccaga ttatatcgcc ccagagataa tcgcttatca gccgtatgga
     5281 aaatctgtgg actggtgggc ctatggcgtc ctgttgtatg aaatgcttgc cgggcagcct
     5341 ccatttgatg gtgaagatga agacgagcta tttcagtcta tcatggagca caacgtttcc
     5401 tatccaaaat ccttgtccaa ggaggctgtt tctatctgca aaggactgat gaccaaacac
     5461 ccagccaagc ggctgggctg tgggcctgag ggggagaggg acgtgagaga gcatgccttc
     5521 ttccggagga tcgactggga aaaactggag aacagggaga tccagccacc attcaagccc
     5581 aaagtgtgtg gcaaaggagc agagaacttt gacaagttct tcacacgagg acagcccgtc
     5641 ttaacaccac ctgatcagct ggttattgct aacatagacc agtctgattt tgaagggttc
     5701 tcgtatgtca acccccagtt tgtgcacccc atcttacaga gtgcagtaga attctatcct
     5761 tacgacgtgc ctgactacgc ctaaggatcc aattctgcag atatccatca cactggcggc
     5821 cgctcgagca tgcatctaga gggccctatt ctatagtgtc acctaaatgc tagagctcgc
     5881 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     5941 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     6001 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     6061 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     6121 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     6181 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     6241 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     6301 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     6361 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     6421 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     6481 ggaacaacac tcaaccctat ctcggtctat tcttttgatt t
//
