LOCUS       sequence-324557-        7212 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 7212)
  AUTHORS   Bingol B, Schuman EM
  TITLE     Activity-dependent dynamics and sequestration of proteasomes in 
            dendritic spines.
  JOURNAL   Nature. 2006 Jun 29. 441(7097):1144-8.
  PUBMED    16810255
REFERENCE   2  (bases 1 to 7212)
  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: "Nature. 
            2006 Jun 29. 441(7097):1144-8."
COMMENT     Sequence Label: pcDNA3.1 CIM5 (Rpt-1) GFP
FEATURES             Location/Qualifiers
     source          1..7212
                     /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"
     CDS             2402..3115
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(2432..2460)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3055..3075
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             3125..3151
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3161..3187
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3220..3237)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3226..3450
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3496..3924
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3583..3602)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3793..3814
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3933..3953)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3938..4267
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4118..4253
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4180..4199
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4315..4362
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4381..4755
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    4885..5006
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4922..4941)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4976..4995
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5055..5071)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5055..5071)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5068..5090)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5079..5095
                     /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(5103..5109,5110..5127,5128..5133))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5148..5169
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5286..5303)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5457..6045)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5537..5556)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6216..7007,7008..7076))
                     /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     6839..6858
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7077..7181)
                     /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 gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattctgc
      961 agatatccag cacagtggcg gccgcgccgc cgccatgcca ccaaaagaag actgggaaaa
     1021 atacaaggca cctttggaag acgatgataa aaaacctgat gatgacaaga tcgtaccctt
     1081 aacagaaggt gacattcaag tcttaaaatc atacggtgca gctccatacg cagccaaatt
     1141 aaagcaaact gagaatgact taaaagacat tgaagcgaga atcaaagaaa aggccggtgt
     1201 gaaagaaagc gatacagggt tagcaccctc ccatctatgg gatattatgg gtgataggca
     1261 gagactgggc gaagaacatc ccttacaagt cgctcgttgc acgaagatca ttaaggggaa
     1321 tggtgaaagt gacgaaacta cgacagataa taataacagc ggcaatagca atagtaatag
     1381 caatcaacag tctactgatg cagacgaaga cgatgaagat gccaagtacg tgataaatct
     1441 taagcaaatt gccaaatttg tcgtcggatt aggtgaacgt gtttctccaa cagatataga
     1501 agaaggtatg cgtgtaggtg tcgataggtc gaaatataat attgaacttc cattgcctcc
     1561 aaggattgac ccatcagtta ctatgatgac cgttgaagaa aagcctgacg ttacttatag
     1621 cgatgtcggt ggctgtaaag accaaattga aaagctgaga gaagttgtcg agctgccctt
     1681 attatctcca gaaagatttg ctactcttgg tattgatcca ccaaagggta tcttattata
     1741 tgggccacct ggtactggta agacattatg tgctcgtgct gttgctaata gaactgatgc
     1801 aacttttatt agggtcattg ggtctgagct agtgcaaaaa tatgtcggtg aaggtgctcg
     1861 tatggttaga gaattatttg aaatggctcg taccaaaaag gcttgcatta tattcttcga
     1921 tgaaatcgac gctgttggtg gtgctcgttt tgatgatggt gctggtggtg ataatgaagt
     1981 tcaaagaact atgttagaac tgattacaca gttagacggg tttgaccctc gtggtaacat
     2041 taaagtgatg tttgccacca ataggcccaa cactttagat ccagctttat tgagacccgg
     2101 tagaattgat cgtaaggtag agttttcatt acctgacttg gaaggtcgtg caaatatctt
     2161 ccgtattcac tctaaatcaa tgagtgtgga acgtggcatc agatgggaac ttatctccag
     2221 actgtgtcca aactctaccg gtgcagaatt aagatctgtt tgcacagaag ccggtatgtt
     2281 tgccattaga gcaagaagaa aggtggctac tgaaaaggat ttcctgaaag ctgttgataa
     2341 agttattagc ggatacaaga agtttagttc cacatcgcgt tatatgcaat ataatggatc
     2401 catgagtaaa ggagaagaac ttttcactgg agttgtccca attcttgttg aattagatgg
     2461 tgatgttaat gggcacaaat tttctgtcag tggagagggt gaaggtgatg caacatacgg
     2521 aaaacttacc cttaaattta tttgcactac tggaaaacta cctgttccat ggccaacact
     2581 tgtcactact ttcacttatg gtgttcaatg cttttcaaga tacccagatc atatgaaacg
     2641 gcatgacttt ttcaagagtg ccatgcccga aggttatgta caggaaagaa ctatattttt
     2701 caaagatgac gggaactaca agacacgtgc tgaagtcaag tttgaaggtg atacccttgt
     2761 taatagaatc gagttaaaag gtattgattt taaagaagat ggaaacattc ttggacacaa
     2821 attggaatac aactataact cacacaatgt atacatcatg gcagacaaac aaaagaatgg
     2881 aatcaaagtt aacttcaaaa ttagacacaa cattgaagat ggaagcgttc aactagcaga
     2941 ccattatcaa caaaatactc caattggcga tggccctgtc cttttaccag acaaccatta
     3001 cctgtccaca caatctgccc tttcgaaaga tcccaacgaa aagagagacc acatggtcct
     3061 tcttgagttt gtaacagctg ctgggattac acatggcatg gatgaactat acaaaggatc
     3121 cgggtaccca tacgatgttc cagattacgc tatgggcgga tacccatatg acgtacctga
     3181 ctatgcagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
     3241 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     3301 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     3361 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     3421 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     3481 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     3541 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     3601 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     3661 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     3721 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     3781 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     3841 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     3901 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     3961 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4021 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4081 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     4141 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     4201 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     4261 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc acgtgttgac
     4321 aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc
     4381 atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc
     4441 gagttctgga ccgaccggct cgggttctcc cgggacttcg tggaggacga cttcgccggt
     4501 gtggtccggg acgacgtgac cctgttcatc agcgcggtcc aggaccaggt ggtgccggac
     4561 aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag
     4621 gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca tgaccgagat cggcgagcag
     4681 ccgtgggggc gggagttcgc cctgcgcgac ccggccggca actgcgtgca cttcgtggcc
     4741 gaggagcagg actgacacgt gctacgagat ttcgattcca ccgccgcctt ctatgaaagg
     4801 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     4861 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     4921 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     4981 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     5041 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     5101 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     5161 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     5221 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     5281 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     5341 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5401 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5461 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5521 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5581 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5641 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5701 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     5761 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     5821 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     5881 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     5941 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     6001 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6061 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6121 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6181 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6241 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6301 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6361 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6421 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6481 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6541 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6601 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     6661 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     6721 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     6781 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     6841 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     6901 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     6961 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     7021 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     7081 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7141 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     7201 ccacctgacg tc
//
