LOCUS Exported 7281 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA3-mouse PKA-RIbeta-mEGFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7281) 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 7281) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7281 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 863..882 /label=T7 /note="T7 promoter, forward primer" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 919..936 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 2098..2817 /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(2143..2164) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2404..2423) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2751..2772 /label=EGFP-C /note="EGFP, forward primer" promoter complement(2833..2851) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(2834..2851) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(2871..2888) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2877..3101 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3147..3575 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3234..3253) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3444..3465 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3584..3604) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3589..3919 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3770..3905 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3832..3851 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3986..4780 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(4040..4059) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4650..4669 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4954..5075 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4991..5010) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5045..5064 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5124..5140) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5124..5140) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5137..5159) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5148..5164 /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(5172..5202) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5217..5238 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5355..5372) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5526..6114) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5606..5625) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6285..7145) /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 6908..6927 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7146..7250) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttgcggcc 901 gccaccatgc ggggttctca tcatcatcat catcatggta tggctagcat ggcctcccca 961 tcatgcttcc actcggagga tgaggactct ctgaaaggat gcgagatgta cgtgcagaaa 1021 catggcatcc agcaggtgct caaagaatgc attgtgcacc tctgtgtcgc caagccggac 1081 cggccactgc gattcctccg ggagcacttt gagaagttgg agaaggagga aaacaggcag 1141 atcctggctc ggcagaagtc aaactcccag tgtgattccc acgatgagga gatctcccca 1201 acacctccaa accccgtggt caaggcgcgt cgccggcggg gcggtgtgag tgctgaagtc 1261 tacactgaag aagatgctgt ctcctacgtg aggaaggtca ttcccaagga ctataagacc 1321 atgaccgcgc tggccaaggc catttctaag aacgtgctct tttctcacct ggacgacaac 1381 gagagaagtg acatatttga cgccatgttt cctgtcactc acatcggtgg ggaaacagtc 1441 atacagcaag ggaatgaagg agataatttc tatgtgattg accaaggaga agtagatgta 1501 tatgtgaacg gggaatgggt gaccaacatc agtgaggggg gaagcttcgg ggagctggct 1561 ctcatctacg gcacccccag agcggctacc gtgaaggcca agacggacct caagctctgg 1621 ggtatcgacc gtgacagcta caggcgcatc ctcatgggaa gcacactgag gaaacgcaag 1681 atgtatgagg agttcctcag caaagtctcc atcctagaat ccctggagaa gtgggaacgc 1741 ctgactgtag ctgatgccct ggagcctgtg cagtttgaag atggagagaa aattgttgtg 1801 cagggggagc ctggagatga cttctacatc atcacagagg gcactgcttc agtcctccag 1861 cgacgatccc ccaatgagga gtacgtggaa gtggggcgcc ttggaccctc tgactacttt 1921 ggggagattg ccctgctgct gaatcggccc cgtgcagcca ctgtggtggc ccggggtccc 1981 ctcaagtgtg tgaagttaga ccggcctcgt tttgagcgtg tcctgggccc ctgctctgag 2041 atcctgaaga ggaacatcca gcgttacaac agcttcatct ccctaactgt cgatatcatg 2101 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc 2161 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc 2221 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc 2281 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag 2341 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc 2401 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg 2461 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag 2521 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc 2581 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac 2641 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac 2701 ctgagcaccc agtccaagct gagcaaagac cccaacgaga agcgcgatca catggtcctg 2761 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaatct 2821 agagggccct attctatagt gtcacctaaa tgctagagct cgctgatcag cctcgactgt 2881 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga 2941 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag 3001 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga 3061 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac 3121 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg 3181 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt 3241 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg 3301 gggcatccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga 3361 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac 3421 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc 3481 tatctcggtc tattcttttg atttataagg gattttgggg atttcggcct attggttaaa 3541 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta 3601 gggtgtggaa agtccccagg ctccccaggc aggcagaagt atgcaaagca tgcatctcaa 3661 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag 3721 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct 3781 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc 3841 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg 3901 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca 3961 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc 4021 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc 4081 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga 4141 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac 4201 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct 4261 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa 4321 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc 4381 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct 4441 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc 4501 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg 4561 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct 4621 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct 4681 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca 4741 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa 4801 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc 4861 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc 4921 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt 4981 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct 5041 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct 5101 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc 5161 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga 5221 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg 5281 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg 5341 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg 5401 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga 5461 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg 5521 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag 5581 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc 5641 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg 5701 ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt 5761 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc 5821 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc 5881 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg 5941 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca 6001 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc 6061 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat 6121 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt 6181 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt 6241 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc 6301 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc 6361 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata 6421 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg 6481 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc 6541 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct 6601 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa 6661 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt 6721 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca 6781 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac 6841 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca 6901 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt 6961 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc 7021 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca 7081 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata 7141 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc 7201 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc 7261 cgaaaagtgc cacctgacgt c //