LOCUS       sequence-173287-        7926 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of Spinophilin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7926)
  AUTHORS   Hiday AC, Edler MC, Salek AB, Morris CW, Thang M, Rentz TJ, Rose KL,
            Jones LM, Baucum AJ 2nd
  TITLE     Mechanisms and Consequences of Dopamine Depletion-Induced 
            Attenuation of the Spinophilin/Neurofilament Medium Interaction.
  JOURNAL   Neural Plast. 2017;2017:4153076. doi: 10.1155/2017/4153076. Epub 
            2017 May 28.
  PUBMED    28634551
REFERENCE   2  (bases 1 to 7926)
  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"; journalName: "Neural 
            Plast."; date: "2017"; volume: "2017:4153076. doi"; pages: " 
            10.1155/2017/4153076. Epub 2017 May 28"
COMMENT     Sequence Label: pcDNA3.1_nHA Spinophilin
FEATURES             Location/Qualifiers
     source          1..7926
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(2222..2246)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2265..2282)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2271..2495
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2541..2969
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2628..2647)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2838..2859
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2978..2998)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2983..3312
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3163..3298
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3225..3244
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3379..4173
                     /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(3433..3452)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4043..4062
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4347..4468
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4384..4403)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4438..4457
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4517..4533)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4517..4533)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4530..4552)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4541..4557
                     /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(4565..4571,4572..4589,4590..4595))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4610..4631
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4748..4765)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4919..5507)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4999..5018)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5678..6469,6470..6538))
                     /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     6301..6320
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6539..6643)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6718..6737)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6909..7288
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7289..7492
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7443..7463
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7537..7556
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7537..7555
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             7588..7614
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             7615..7635
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    7666..7690
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 ggcctggccg aggccccacg ggcgtccgag cgcggcgtgc gcctgtcgct gccgcgggcc
       61 agcagcctga acgagaacgt ggaccacagc gccctgctga agctgggcac cagcgtgtcg
      121 gagcgcgtga gccgcttcga ctccaagccc gcgccctccg cgcagcctgc gccgccgccg
      181 cacccgccgt cccggctgca ggagacgcgg aagctgttcg aacggagcgc cccagcggcc
      241 gcaggcggcg acaaggaggc cgcggcgcgg cggctgctga ggcaggagcg cgccggcctg
      301 caggaccgga agctggacgt cgtggtgcgc ttcaacggca gcaccgaggc gctggacaag
      361 ctggacgctg acgccgtgtc ccccacggtc agccagctca gcgccgtctt cgagaaggcc
      421 gactcgagga ccggcctcca ccgcgggccc gggctcccca gggccgcagg ggttccccag
      481 gtcaactcga agctggtcag caagcggtcc cgggtgttcc agcccccgcc gccgccgccg
      541 cccgccccgt cgggggatgc cccggccgag aaagagcgat gccccgcagg gcagcagccc
      601 ccgcagcacc gagtggcccc tgcccggccg ccccccaagc cccgggaggt gcgcaagatt
      661 aagccggtgg aggtggagga gagcggggag tcggaggccg agtcggcgcc cggggaggta
      721 atccaggccg aggttacggt ccacgcggcc ctggagaatg gcagcaccgt ggcaactgca
      781 gccagccccg cgcccgagga gccaaaggcc caagcggccc cggagaagga ggcggcggcg
      841 gtagcgccgc cagagagggg ggtgggcaat ggccgggccc cggacgtggc ccctgaggag
      901 gtagatgaat ccaagaagga ggacttctcg gaggcggact tggtggacgt gagcgcctac
      961 agtgggctcg gggaggactc tgcgggcagt gccctggagg aggacgacga agacgacgag
     1021 gaggatgggg agccccccta cgagcccgag tcggggtgcg tggagatccc ggggctgtcg
     1081 gaggaggagg acccagcccc gagccggaag atccatttca gcacggcgcc catccaagtg
     1141 ttcagcactt actccaacga ggattacgat cgtcgcaacg aggatgtgga tcccatggca
     1201 gcctctgctg agtacgagct ggagaagcgt gtggagaggt tggagctgtt ccctgtggag
     1261 ctggagaagg actccgaggg cctgggcatc agcatcatcg gcatgggcgc cggggcagac
     1321 atgggcctgg agaagctggg tatcttcgtc aagaccgtga cggagggtgg tgcggcccat
     1381 cgggatggca ggatccaggt gaatgatctc ctggtggagg tggatggaac aagtctggtg
     1441 ggagtgaccc agagcttcgc ggcgtctgtg ctccggaaca ccaagggccg agtgcggttt
     1501 atgattggcc gggagcggcc gggagagcag agcgaagtgg cccagctaat tcagcagact
     1561 ttggaacagg agcgatggca gcgggagatg atggagcaga gatacgccca gtatggggag
     1621 gatgacgagg agacgggaga gtatgccact gacgaggatg aggagctgag ccccacgttc
     1681 ccgggtggtg agatggccat cgaggtgttt gagctagcgg agaacgagga tgcactgtcc
     1741 cctgtggaca tggagcccga gaagctggtg cacaagttca aggagctcca gatcaagcat
     1801 gcggtcactg aggcagagat ccagcagctg aaaagaaagc tgcagagcct ggagcaggag
     1861 aaggggcgct ggcgggtgga gaaggcgcag ttggagcaga gtgtggagga gaacaaggag
     1921 cgcatggaga aactggaagg ctactggggt gaggcccaga gcctgtgcca ggctgtggac
     1981 gagcacctgc gggagactca ggcgcagtac caggccctgg agcgcaagta cagcaaggcc
     2041 aagcgcctca tcaaggacta ccagcagaag gagatcgagt tcctgaaaaa ggagactgca
     2101 cagcgtcggg ttctggagga gtcggagctg gccagaaagg aggagatgga caagctcctg
     2161 gacaagatct cagaactgga aggaaacttg caaacactga ggaattccaa ttctacttaa
     2221 tacccagctt tcttgtacaa agtggtgata aacccgctga tcagcctcga ctgtgccttc
     2281 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     2341 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     2401 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     2461 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     2521 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2581 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     2641 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     2701 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     2761 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     2821 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     2881 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     2941 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     3001 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3061 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3121 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     3181 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     3241 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     3301 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     3361 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     3421 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     3481 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     3541 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     3601 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     3661 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     3721 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     3781 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     3841 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     3901 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     3961 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     4021 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     4081 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     4141 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     4201 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     4261 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     4321 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     4381 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     4441 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     4501 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     4561 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     4621 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     4681 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     4741 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     4801 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     4861 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4921 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4981 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     5041 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5101 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5161 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5221 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5281 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5341 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5401 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5461 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5521 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5581 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5641 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     5701 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     5761 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     5821 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     5881 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     5941 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     6001 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6061 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6121 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6181 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6241 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6301 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6361 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6421 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6481 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6541 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6601 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6661 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac
     6721 aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt
     6781 cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg
     6841 catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata
     6901 tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
     6961 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     7021 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     7081 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     7141 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     7201 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     7261 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
     7321 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
     7381 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
     7441 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctctctggct
     7501 aactagagaa cccactgctt actggcttat cgaaattaat acgactcact atagggagac
     7561 ccaagctggc tagttaagct taccatgtac ccatacgatg ttccagatta cgctgaaaac
     7621 ctgtattttc agggcccgga tccgtcgacg aattctgcag atatcacaag tttgtacaaa
     7681 aaagcaggct taatgatgaa gacggagcca cgggggcccg ggggtcccct ccggagcgcc
     7741 tccccgcacc gcagcgccta cgaggcgggc atccaggcgc tgaagccgcc cgacgcgccc
     7801 gggcccgacg aggcacccaa gggggcccac cacaagaaat atggctccaa cgtccaccgc
     7861 atcaaaagta tgttcctgca gatgggcacg acggcggggc cctcgggcga ggcgggcggc
     7921 ggcgcg
//
