LOCUS       sequence-63624-e        7745 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 7745)
  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 7745)
  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-mouse PKA-Calpha-mEGFP
FEATURES             Location/Qualifiers
     source          1..7745
                     /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             2808..2834
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     CDS             join(2835..2837,2838..3551)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2877..2898)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3138..3157)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3485..3506
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     intron          3668..3801
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3784..3803
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    3810..4398
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3863..3883)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4281..4292)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4425..4442)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4431..4542
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4769..4788)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4834..4889
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4888..4907)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5248..5264)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5261..5283)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5272..5288
                     /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(5296..5302,5303..5320,5321..5326))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5341..5362
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5421..5616
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5467..5602
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5622..5756
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5659..5678)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5713..5732
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5824..5841)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5995..6583)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6075..6094)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6754..7545,7546..7614))
                     /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     7377..7396
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7615..7719)
                     /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 ccgctaccat gggcaacgcc gccgccgcca agaagggcag cgagcaggag agcgtgaaag
     1801 agttcctagc caaagccaag gaagatttcc tgaaaaaatg ggagacccct tctcagaata
     1861 cagcccagtt ggatcagttt gatagaatca agacccttgg caccggctcc tttgggcgag
     1921 tgatgctggt gaagcacaag gagagtggga accactacgc catgaagatc ttagacaagc
     1981 agaaggtggt gaagctaaag cagatcgagc acactctgaa tgagaagcgc atcctgcagg
     2041 ccgtcaactt cccgttcctg gtcaaacttg aattctcctt caaggacaac tcaaacctgt
     2101 acatggtcat ggagtatgta gctggtggcg agatgttctc ccacctacgg cggattggaa
     2161 ggttcagcga gccccatgcc cgtttctacg cggcgcagat cgtcctgacc tttgagtatc
     2221 tgcactccct ggacctcatc taccgggacc tgaagcccga gaatcttctc atcgaccagc
     2281 agggctatat tcaggtgaca gacttcggtt ttgccaagcg tgtgaaaggc cgtacttgga
     2341 ccttgtgtgg gacccctgag tacttggccc ccgagattat cctgagcaaa ggctacaaca
     2401 aggctgtgga ctggtgggct ctcggagtcc tcatctacga gatggctgct ggttacccac
     2461 ccttcttcgc tgaccagcct atccagatct atgagaaaat cgtctctggg aaggtgcggt
     2521 tcccatccca cttcagctct gacttgaagg acctgctgcg gaaccttctg caggtggatc
     2581 tcaccaagcg ctttgggaac ctcaagaacg gggtcaatga catcaagaac cacaagtggt
     2641 ttgccacgac tgactggatt gccatctatc agagaaaggt ggaagctccc ttcataccaa
     2701 agtttaaagg ccctggggac acgagtaact ttgacgacta tgaggaggaa gagatccggg
     2761 tctccatcaa tgagaagtgt ggcaaggagt ttactgagtt tatcgattat gatgttcctg
     2821 attatgcaag cttaatgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     2881 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     2941 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     3001 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
     3061 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     3121 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     3181 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     3241 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     3301 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
     3361 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     3421 ccgacaacca ctacctgagc acccagtcca agctgagcaa agaccccaac gagaagcgcg
     3481 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     3541 tgtacaagta atctagggga tccaccggaa gccacctgca tgcggccgca tgatcagctg
     3601 gatgcatcga tcacgcgtac cggtgctcga ggtaccgatg aatagctaag gtcgaccgag
     3661 gccgcaggta agtatcaagg ttacaagaca ggtttaagga gaccacatag aaactgggct
     3721 tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc
     3781 actttgcctt tctctccaca ggtgtcgaca atcaacctct ggattacaaa atttgtgaaa
     3841 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     3901 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     3961 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     4021 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     4081 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     4141 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     4201 ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     4261 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     4321 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     4381 tttgggccgc ctccccgcct ggaattcgag ctcggtacga tcagcctcga ctgtgccttc
     4441 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     4501 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     4561 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     4621 tagcaggcca gcttttgttc cctttagggt taattgcgcg cttggcgcta atcatggtca
     4681 tagctgtttc cactcctcag gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca
     4741 atgccctggc tcacaaatac cactgagatc tttttccctc tgccaaaaat tatggggaca
     4801 tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt ttcattgcaa
     4861 tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt
     4921 taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgccata tgctggctgc
     4981 catgaacaaa ggtggctata aagaggtcat cagtatatga aacagccccc tgctgtccat
     5041 tccttattcc atagaaaagc cttgacttga ggttagattt ttttttatat tttgtttgtg
     5101 ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc
     5161 ctcctctcct gactactccc agtcatagct gtccctcttc tcttatgaag atccctcgac
     5221 ctgcagccca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     5281 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     5341 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     5401 cctgtcgtgc cagcggatcc gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     5461 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     5521 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     5581 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc taacttgttt attgcagctt
     5641 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     5701 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatccgct
     5761 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5821 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5881 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5941 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     6001 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     6061 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     6121 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     6181 gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     6241 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     6301 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6361 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6421 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     6481 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     6541 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6601 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6661 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6721 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     6781 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     6841 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     6901 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     6961 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     7021 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     7081 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     7141 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     7201 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     7261 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     7321 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     7381 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     7441 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     7501 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     7561 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     7621 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     7681 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     7741 acctg
//
