LOCUS       sequence-63618-e        5972 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 5972)
  AUTHORS   Zhong H, Sia GM, Sato TR, Gray NW, Mao T, Khuchua Z, Huganir RL, 
            Svoboda K
  TITLE     Subcellular dynamics of type II PKA in neurons.
  JOURNAL   Neuron. 2009 May 14;62(3):363-74. doi: 10.1016/j.neuron.2009.03.013.
  PUBMED    19447092
REFERENCE   2  (bases 1 to 5972)
  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"; doi: 
            "10.1016/j.neuron.2009.03"; journalName: "Neuron"; date: 
            "2009-05-14- 14"; volume: "62"; issue: "3"; pages: "363-74"
COMMENT     Sequence Label: pRK5-rat PKA-RIIbeta
FEATURES             Location/Qualifiers
     source          1..5972
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        14..393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        394..597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     548..568
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     594..618
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        828..846
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     828..845
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     859..878
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      2221..2230
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2234..2368
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2271..2290)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2325..2344
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        2437..2794
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2437..2628
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2460..2480)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2645..2780
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2814..2831)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2814..2830)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2823..2845)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3043..3498
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3130..3149)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3340..3361
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3515..3534)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3669..3691
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3744..3762)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3830..3934
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3935..4003,4004..4795)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRDDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4153..4172)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4966..5554
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5455..5474
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5708..5725
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(5878..5883,5884..5901,5902..5908)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5916..5932
                     /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     5921..5943
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5940..5956
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5940..5956
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ttcgagctcg cccgacattg attattgact agttattaat agtaatcaat tacggggtca
       61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      361 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      421 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      481 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      541 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
      601 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
      661 caccgggacc gatccagcct ccgcggccgg gaacggtgca ttggaacgcg gattccccgt
      721 gccaagagtg acgtaagtac cgcctataga gtctataggc ccaccccctt ggcttcgtta
      781 gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt taggtgacac
      841 tatagaataa catccacttt gcctttctct ccacaggtgt ccactcccag gtccaactgc
      901 acctcggttc tatcgattga attcgtcgcc accatgagca tcgagatccc cgcggggctc
      961 acggagctgc tgcagggctt cacggtggag gtgctgaggc accagcccgc cgacctgctg
     1021 gagttcgcgc tgcagcactt cacgcggctg cagcaggaga acgagcgcaa gggcgccgcg
     1081 cgcttcggcc atgagggcag gacctggggg gacgcgggcg cagccgccgg gggcggaacc
     1141 cccagtaagg gtgtcaactt cgcggaggag cccatgcgct ccgattccga gaacggcgaa
     1201 gaggaggagg ccgcggaagc aggggcgttc aacgctccag ttataaaccg gttcacaagg
     1261 cgtgcctcgg tatgtgcaga agcttataat cctgatgaag aagaagatga tgcagagtcc
     1321 aggataatac atcccaaaac tgacgatcaa agaaacagat tgcaagaagc ctgcaaagac
     1381 atcctgctgt ttaagaacct ggatccagaa cagatgtctc aagtattaga tgccatgttt
     1441 gaaaaattgg tcaaagaagg ggaacacgta atcgatcaag gtgatgatgg tgacaacttt
     1501 tacgtcatcg acagaggaac atttgatatt tatgtaaaat gtgatggcgt tggaagatgc
     1561 gttggtaact atgacaatcg tgggagtttt ggagaactgg ccttaatgta caatacaccc
     1621 agagcagcta caatcactgc tacctctcct ggtgctctgt ggggtttgga cagggtgacc
     1681 ttcaggagaa taatagtaaa aaacaatgcc aaaaagagga agatgtacga gagctttatt
     1741 gagtcactgc cattcctcaa gtctctggag gtttctgaac gcctgaaggt ggtagatgtg
     1801 attggcacca aagtttacaa cgatggagaa cagatcattg ctcagggaga ctcggcggat
     1861 tcgttcttca ttgtagaatc tggagaagtg agaattacta tgaagagaaa gggtaaatca
     1921 gacatcgaag agaacggtgc tgtggaaatc gctcggtgtc tccggggaca gtattttgga
     1981 gagcttgccc tggtcactaa caagccaaga gcagcatctg cacacgccat tgggactgtc
     2041 aaatgcttag ccatggatgt gcaagcattt gagaggcttc tgggaccttg catggaaatt
     2101 atgaagagga atatcgccac ctatgaggag caattagttg cactgtttgg gacaaacatg
     2161 gatattgttg agcccactgc ataagcggcc gcgactctag agtcgacctg cagaagcttg
     2221 gccgccatgg cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     2281 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     2341 catcaatgta tcttatcatg tctggatcgg gaattaattc ggcgcagcac catggcctga
     2401 aataacctct gaaagaggaa cttggttagg taccttctga ggcggaaaga accagctgtg
     2461 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     2521 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     2581 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     2641 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     2701 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     2761 aggaggcttt tttggaggcc taggcttttg caaaaagctg ttaacagctt ggcactggcc
     2821 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     2881 gcacatcccc ccttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     2941 caacagttgc gtagcctgaa tggcgaatgg cgcctgatgc ggtattttct ccttacgcat
     3001 ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg
     3061 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     3121 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     3181 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     3241 accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     3301 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     3361 gaacaacact caaccctatc tcgggctatt cttttgattt ataagggatt ttgccgattt
     3421 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     3481 tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     3541 taagccaact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac
     3601 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     3661 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     3721 gcagtattct tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     3781 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     3841 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3901 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     3961 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     4021 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     4081 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     4141 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgat gacgccgggc aagagcaact
     4201 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     4261 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     4321 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     4381 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     4441 agccatacca aacgacgagc gtgacaccac gatgccagca gcaatggcaa caacgttgcg
     4501 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     4561 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     4621 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     4681 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     4741 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     4801 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     4861 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     4921 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4981 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5041 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5101 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5161 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5221 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5281 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5341 atacctacag cgtgagcatt gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     5401 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     5461 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     5521 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     5581 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     5641 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     5701 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     5761 ccccgcgcgt tggccgattc attaatccag ctggcacgac aggtttcccg actggaaagc
     5821 gggcagtgag cgcaacgcaa ttaatgtgag ttacctcact cattaggcac cccaggcttt
     5881 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     5941 aggaaacagc tatgaccatg attacgaatt aa
//
