LOCUS       sequence-63626-e        7916 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 7916)
  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 7916)
  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: pCAGGS-rat PKA-RIIbeta-mEGFP
FEATURES             Location/Qualifiers
     source          1..7916
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1670
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1594..1616
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             join(3003..3005,3006..3008,3009..3722)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3048..3069)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3309..3328)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3656..3677
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     intron          3839..3972
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3955..3974
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    3981..4569
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4034..4054)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4452..4463)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4596..4613)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4602..4713
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4940..4959)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5005..5060
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5059..5078)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5419..5435)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5432..5454)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5443..5459
                     /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(5467..5473,5474..5491,5492..5497))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5512..5533
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5592..5787
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5638..5773
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5793..5927
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5830..5849)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5884..5903
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5995..6012)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6166..6754)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6246..6265)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6925..7716,7717..7785))
                     /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     7548..7567
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7786..7890)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg ggcgagggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
     1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
     1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
     1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
     1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     1681 acgtgctggt tgttgtgctg tctcatcatt ttggcaaaga attcggtacc gcgggcccgg
     1741 ccgccaccat gagcatcgag atccccgcgg ggctcacgga gctgctgcag ggcttcacgg
     1801 tggaggtgct gaggcaccag cccgccgacc tgctggagtt cgcgctgcag cacttcacgc
     1861 ggctgcagca ggagaacgag cgcaagggcg ccgcgcgctt cggccatgag ggcaggacct
     1921 ggggggacgc gggcgcagcc gccgggggcg gaacccccag taagggtgtc aacttcgcgg
     1981 aggagcccat gcgctccgat tccgagaacg gcgaagagga ggaggccgcg gaagcagggg
     2041 cgttcaacgc tccagttata aaccggttca caaggcgtgc ctcggtatgt gcagaagctt
     2101 ataatcctga tgaagaagaa gatgatgcag agtccaggat aatacatccc aaaactgacg
     2161 atcaaagaaa cagattgcaa gaagcctgca aagacatcct gctgtttaag aacctggatc
     2221 cagaacagat gtctcaagta ttagatgcca tgtttgaaaa attggtcaaa gaaggggaac
     2281 acgtaatcga tcaaggtgat gatggtgaca acttttacgt catcgacaga ggaacatttg
     2341 atatttatgt aaaatgtgat ggcgttggaa gatgcgttgg taactatgac aatcgtggga
     2401 gttttggaga actggcctta atgtacaata cacccagagc agctacaatc actgctacct
     2461 ctcctggtgc tctgtggggt ttggacaggg tgaccttcag gagaataata gtaaaaaaca
     2521 atgccaaaaa gaggaagatg tacgagagct ttattgagtc actgccattc ctcaagtctc
     2581 tggaggtttc tgaacgcctg aaggtggtag atgtgattgg caccaaagtt tacaacgatg
     2641 gagaacagat cattgctcag ggagactcgg cggattcgtt cttcattgta gaatctggag
     2701 aagtgagaat tactatgaag agaaagggta aatcagacat cgaagagaac ggtgctgtgg
     2761 aaatcgctcg gtgtctccgg ggacagtatt ttggagagct tgccctggtc actaacaagc
     2821 caagagcagc atctgcacac gccattggga ctgtcaaatg cttagccatg gatgtgcaag
     2881 catttgagag gcttctggga ccttgcatgg aaattatgaa gaggaatatc gccacctatg
     2941 aggagcaatt agttgcactg tttgggacaa acatggatat tgttgagccc actgcagata
     3001 tcatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     3061 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     3121 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     3181 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
     3241 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     3301 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     3361 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     3421 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     3481 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     3541 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     3601 actacctgag cacccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg
     3661 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     3721 aatctagggg atccaccgga agccacctgc atgcggccgc atgatcagct ggatgcatcg
     3781 atcacgcgta ccggtgctcg aggtaccgat gaatagctaa ggtcgaccga ggccgcaggt
     3841 aagtatcaag gttacaagac aggtttaagg agaccacata gaaactgggc ttgtcgagac
     3901 agagaagact cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct
     3961 ttctctccac aggtgtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg
     4021 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     4081 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     4141 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     4201 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     4261 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     4321 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga
     4381 cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     4441 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     4501 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     4561 cctccccgcc tggaattcga gctcggtacg atcagcctcg actgtgcctt ctagttgcca
     4621 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     4681 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     4741 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggcc
     4801 agcttttgtt ccctttaggg ttaattgcgc gcttggcgct aatcatggtc atagctgttt
     4861 ccactcctca ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc aatgccctgg
     4921 ctcacaaata ccactgagat ctttttccct ctgccaaaaa ttatggggac atcatgaagc
     4981 cccttgagca tctgacttct ggctaataaa ggaaatttat tttcattgca atagtgtgtt
     5041 ggaatttttt gtgtctctca ctcggaagga catatgggag ggcaaatcat ttaaaacatc
     5101 agaatgagta tttggtttag agtttggcaa catatgccat atgctggctg ccatgaacaa
     5161 aggtggctat aaagaggtca tcagtatatg aaacagcccc ctgctgtcca ttccttattc
     5221 catagaaaag ccttgacttg aggttagatt tttttttata ttttgtttgt gttatttttt
     5281 tctttaacat ccctaaaatt ttccttacat gttttactag ccagattttt cctcctctcc
     5341 tgactactcc cagtcatagc tgtccctctt ctcttatgaa gatccctcga cctgcagccc
     5401 aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     5461 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     5521 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     5581 ccagcggatc cgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     5641 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     5701 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     5761 ttttttggag gcctaggctt ttgcaaaaag ctaacttgtt tattgcagct tataatggtt
     5821 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5881 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatccgc tgcattaatg
     5941 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     6001 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     6061 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     6121 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     6181 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     6241 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     6301 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     6361 atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     6421 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     6481 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     6541 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     6601 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     6661 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     6721 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     6781 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     6841 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     6901 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     6961 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     7021 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     7081 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     7141 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     7201 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     7261 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     7321 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     7381 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     7441 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     7501 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     7561 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     7621 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     7681 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     7741 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     7801 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     7861 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctg
//
