LOCUS       sequence-307416-        7697 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes residues 1-469 of rat soluble adenylyl cyclase (ADCY10) 
            fused to mCherry; nuclear targeted..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7697)
  AUTHORS   Sample V, DiPilato LM, Yang JH, Ni Q, Saucerman JJ, Zhang J
  TITLE     Regulation of nuclear PKA revealed by spatiotemporal manipulation of
            cyclic AMP.
  JOURNAL   Nat Chem Biol. 2012 Feb 26;8(4):375-82. doi: 10.1038/nchembio.799.
  PUBMED    22366721
REFERENCE   2  (bases 1 to 7697)
  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: "10.1038/nchembio"; 
            journalName: "Nat Chem Biol"; date: "2012-02-26- 26"; volume: "8"; 
            issue: "4"; pages: "375-82"
COMMENT     Sequence Label: pcDNA3-sACt-mCherry-NLS
FEATURES             Location/Qualifiers
     source          1..7697
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     regulatory      901..910
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             922..939
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             943..975
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(979..987,988..1002)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             2449..3156
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2594..2612)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2862..2881
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             3157..3177
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(3250..3268)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3251..3268)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3288..3305)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3294..3518
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3564..3992
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3651..3670)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3861..3882
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4001..4021)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4006..4335
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4186..4321
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4248..4267
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4402..5196
                     /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(4456..4475)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5066..5085
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5370..5491
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5407..5426)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5461..5480
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5540..5556)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5540..5556)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5553..5575)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5564..5580
                     /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(5588..5594,5595..5612,5613..5618))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5633..5654
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5771..5788)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5942..6530)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6022..6041)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6701..7492,7493..7561))
                     /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     7324..7343
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7562..7666)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct 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 atgggtggag 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 gccaccatgg tgcggggttc tcatcatcat catcatcatg gtatggctag catgactggt
      961 ggacagcaaa tgggtcggga tctgtacgac gatgacgata aggatccgag ctcgagatct
     1021 gcagctgcca tggacatgag tgcccgaagg caggaattac aggacagggc aatcgtcaag
     1081 atagctgctc acttaccgga cctcattgtc tatggagatt tctctcccga gcggccgtca
     1141 gtgaaatgtt ttgatggagt tctgatgttt gtcgatattt caggctttac tgcaatgact
     1201 gagaagttca gcacagccat gtacatggac cgaggagccg agcagctggt ggagatcctc
     1261 aactactaca taagtgcgat agtggagaaa gtactgattt ttggaggaga catcctaaaa
     1321 tttgcaggtg acgccttgtt ggccctgtgg aaagtggaac gaaagcaact gaagaatatc
     1381 atcacggtgg taattaaatg cagcctggag attcatggct tgtttgaagc caaggaggtt
     1441 gaagaaggcc tggatattcg agttaagata ggactggctg ctggccacat caccatgttg
     1501 gtctttgggg atgaaacacg gaactacttc ctggtgattg gccaagcggt ggatgatgta
     1561 cgccttgctc agaacatggc tcagatgaat gatgttattc tgtcaccaaa ctgctggcag
     1621 ctctgtgatc ggagcatgat tgaaatcgag aggattccgg atcagagagc agttaaggtt
     1681 agcttcttaa aaccaccccc aacttttaac ttcgacgagt tttttgccaa gtgtatggcc
     1741 ttcatggatt attatccttc tggtgaccac aaaaacttcc taaggcttgc ctgcatgctg
     1801 gagtctgatc ctgaactcga gttgtctcta caaaagtatg tgatggaaat cattttgaag
     1861 cagattgatg acaagcagct tcggggctat ttatctgagc ttcgtcctgt gacgatcgtg
     1921 tttgtgaact tgatgtttaa agagcaagac aaagcagaag tcataggatc agccatccaa
     1981 gctgcctgtg tgcacatcac ttccgtcttg aaggtcttcc gaggccagat caataaggtc
     2041 ttcatgtttg ataagggctg ctccttcctc tgtgtcttcg gtttccctgg ggaaaaggcc
     2101 cctgacgaga tcactcacgc tttggaaagt gccgtggata tattcgactt ctgctctcag
     2161 gtccacaaaa tccgtactgt ctccatcggc gtcgccagtg ggattgtctt ctgtgggatc
     2221 gttggacaca ctgtgagaca cgagtacaca gtcattggcc aaaaggtcaa tattgctgcc
     2281 aggatgatga tgtattaccc aggcatcgtg acctgcgact ctgtcacata cgatggcagc
     2341 aacctgccag cctacttttt taaagagctt ccaaagaaag tcatgaaagg agttgcggac
     2401 cccggaccag tgtatcagtg tctgggcctc aatgagaaag tcgtcgacat ggtgagcaag
     2461 ggcgaagagg ataacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag
     2521 ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag
     2581 ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac
     2641 atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc cgccgacatc
     2701 cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc
     2761 gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc
     2821 tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag
     2881 accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc cctgaagggc
     2941 gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga ggtcaagacc
     3001 acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa catcaagttg
     3061 gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg cgccgagggc
     3121 cgccactcca ccggcggcat ggacgagctg tacaagccaa aaaagaagag aaaggtggaa
     3181 gacgcataag aattctgcag atatccatca cactggcggc cgctcgagca tgcatctaga
     3241 gggccctatt ctatagtgtc acctaaatgc tagagctcgc tgatcagcct cgactgtgcc
     3301 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     3361 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     3421 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     3481 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     3541 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3601 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     3661 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3721 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     3781 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3841 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     3901 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     3961 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     4021 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4081 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4141 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     4201 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     4261 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     4321 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     4381 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     4441 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     4501 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     4561 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     4621 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     4681 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     4741 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     4801 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     4861 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     4921 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     4981 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     5041 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     5101 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     5161 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     5221 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     5281 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     5341 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     5401 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     5461 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     5521 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     5581 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     5641 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     5701 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     5761 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     5821 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     5881 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     5941 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     6001 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     6061 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     6121 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     6181 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     6241 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     6301 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     6361 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     6421 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     6481 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     6541 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     6601 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     6661 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     6721 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     6781 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6841 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6901 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6961 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     7021 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     7081 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     7141 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     7201 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     7261 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     7321 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     7381 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     7441 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     7501 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     7561 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     7621 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     7681 aagtgccacc tgacgtc
//
