LOCUS       sequence-63625-e        7201 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 7201)
  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 7201)
  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
FEATURES             Location/Qualifiers
     source          1..7201
                     /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..1673
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1597..1619
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1681..1700
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          3124..3257
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3240..3259
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    3266..3854
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3319..3339)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3737..3748)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3881..3898)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3887..3998
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4225..4244)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4290..4345
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4344..4363)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4704..4720)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4717..4739)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4728..4744
                     /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(4752..4758,4759..4776,4777..4782))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4797..4818
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4877..5072
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4923..5058
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5078..5212
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5115..5134)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5169..5188
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5280..5297)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5451..6039)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5531..5550)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6210..7001,7002..7070))
                     /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     6833..6852
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7071..7175)
                     /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 tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct
     1441 ctagcgggcg cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc
     1501 ttcgtgcgtc gccgcgccgc cgtccccttc tccctctcca gcctcggggc tgtccgcggg
     1561 gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg
     1621 gcggctctag agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg
     1681 gcaacgtgct ggttattgtg ctgtctcatc attttggcaa agaattcggt accgcgggcc
     1741 caattcgtcg ccaccatgag catcgagatc cccgcggggc tcacggagct gctgcagggc
     1801 ttcacggtgg aggtgctgag gcaccagccc gccgacctgc tggagttcgc gctgcagcac
     1861 ttcacgcggc tgcagcagga gaacgagcgc aagggcgccg cgcgcttcgg ccatgagggc
     1921 aggacctggg gggacgcggg cgcagccgcc gggggcggaa cccccagtaa gggtgtcaac
     1981 ttcgcggagg agcccatgcg ctccgattcc gagaacggcg aagaggagga ggccgcggaa
     2041 gcaggggcgt tcaacgctcc agttataaac cggttcacaa ggcgtgcctc ggtatgtgca
     2101 gaagcttata atcctgatga agaagaagat gatgcagagt ccaggataat acatcccaaa
     2161 actgacgatc aaagaaacag attgcaagaa gcctgcaaag acatcctgct gtttaagaac
     2221 ctggatccag aacagatgtc tcaagtatta gatgccatgt ttgaaaaatt ggtcaaagaa
     2281 ggggaacacg taatcgatca aggtgatgat ggtgacaact tttacgtcat cgacagagga
     2341 acatttgata tttatgtaaa atgtgatggc gttggaagat gcgttggtaa ctatgacaat
     2401 cgtgggagtt ttggagaact ggccttaatg tacaatacac ccagagcagc tacaatcact
     2461 gctacctctc ctggtgctct gtggggtttg gacagggtga ccttcaggag aataatagta
     2521 aaaaacaatg ccaaaaagag gaagatgtac gagagcttta ttgagtcact gccattcctc
     2581 aagtctctgg aggtttctga acgcctgaag gtggtagatg tgattggcac caaagtttac
     2641 aacgatggag aacagatcat tgctcaggga gactcggcgg attcgttctt cattgtagaa
     2701 tctggagaag tgagaattac tatgaagaga aagggtaaat cagacatcga agagaacggt
     2761 gctgtggaaa tcgctcggtg tctccgggga cagtattttg gagagcttgc cctggtcact
     2821 aacaagccaa gagcagcatc tgcacacgcc attgggactg tcaaatgctt agccatggat
     2881 gtgcaagcat ttgagaggct tctgggacct tgcatggaaa ttatgaagag gaatatcgcc
     2941 acctatgagg agcaattagt tgcactgttt gggacaaaca tggatattgt tgagcccact
     3001 gcataagcgg ccgggatcca ccggaagcca cctgcatgcg gccgcatgat cagctggatg
     3061 catcgatcac gcgtaccggt gctcgaggta ccgatgaata gctaaggtcg accgaggccg
     3121 caggtaagta tcaaggttac aagacaggtt taaggagacc acatagaaac tgggcttgtc
     3181 gagacagaga agactcttgc gtttctgata ggcacctatt ggtcttactg acatccactt
     3241 tgcctttctc tccacaggtg tcgacaatca acctctggat tacaaaattt gtgaaagatt
     3301 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     3361 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     3421 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     3481 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     3541 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     3601 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     3661 gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     3721 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     3781 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     3841 ggccgcctcc ccgcctggaa ttcgagctcg gtacgatcag cctcgactgt gccttctagt
     3901 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     3961 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     4021 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     4081 aggccagctt ttgttccctt tagggttaat tgcgcgcttg gcgctaatca tggtcatagc
     4141 tgtttccact cctcaggtgc aggctgccta tcagaaggtg gtggctggtg tggccaatgc
     4201 cctggctcac aaataccact gagatctttt tccctctgcc aaaaattatg gggacatcat
     4261 gaagcccctt gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt
     4321 gtgttggaat tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa
     4381 acatcagaat gagtatttgg tttagagttt ggcaacatat gccatatgct ggctgccatg
     4441 aacaaaggtg gctataaaga ggtcatcagt atatgaaaca gccccctgct gtccattcct
     4501 tattccatag aaaagccttg acttgaggtt agattttttt ttatattttg tttgtgttat
     4561 ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga tttttcctcc
     4621 tctcctgact actcccagtc atagctgtcc ctcttctctt atgaagatcc ctcgacctgc
     4681 agcccaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     4741 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     4801 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     4861 tcgtgccagc ggatccgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     4921 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     4981 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     5041 gaggcttttt tggaggccta ggcttttgca aaaagctaac ttgtttattg cagcttataa
     5101 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     5161 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga tccgctgcat
     5221 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     5281 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     5341 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     5401 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     5461 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5521 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     5581 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     5641 tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5701 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5761 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5821 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5881 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5941 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     6001 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     6061 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     6121 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     6181 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     6241 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     6301 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     6361 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     6421 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     6481 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     6541 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     6601 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     6661 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     6721 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     6781 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     6841 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     6901 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     6961 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     7021 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     7081 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     7141 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     7201 g
//
