LOCUS       sequence-8824-ex        5574 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 5574)
  AUTHORS   Harvey CD, Yasuda R, Zhong H, Svoboda K
  TITLE     The spread of Ras activity triggered by activation of a single 
            dendritic spine.
  JOURNAL   Science. 2008 Jul 4. 321(5885):136-40.
  PUBMED    18556515
REFERENCE   2  (bases 1 to 5574)
  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: "Science. 
            2008 Jul 4. 321(5885):136-40."
COMMENT     Sequence Label: MARCKS(1-40)-paGFP
FEATURES             Location/Qualifiers
     source          1..5574
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        14..393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        394..597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     548..568
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     594..618
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        828..846
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     828..845
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     859..878
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             join(1081..1083,1084..1086,1087..1797)
                     /codon_start=1
                     /product="photoactivatable GFP variant (Patterson et al., 
                     2002)"
                     /label=PA-GFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1126..1147)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1387..1406)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1734..1755
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      1819..1828
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1832..1966
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1869..1888)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1923..1942
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        2039..2396
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2039..2230
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2062..2082)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2247..2382
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2416..2433)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2416..2432)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2425..2447)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2645..3100
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2732..2751)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2942..2963
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3117..3136)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3271..3293
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3346..3364)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3432..3536
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3537..3605,3606..4397)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRDDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3755..3774)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4568..5156
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5057..5076
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5310..5327
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(5480..5485,5486..5503,5504..5510)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5518..5534
                     /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)."
     primer_bind     5523..5545
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5542..5558
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5542..5558
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ttcgagctcg cccgacattg attattgact agttattaat agtaatcaat tacggggtca
       61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      361 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      421 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      481 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      541 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
      601 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
      661 caccgggacc gatccagcct ccgcggccgg gaacggtgca ttggaacgcg gattccccgt
      721 gccaagagtg acgtaagtac cgcctataga gtctataggc ccaccccctt ggcttcgtta
      781 gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt taggtgacac
      841 tatagaataa catccacttt gcctttctct ccacaggtgt ccactcccag gtccaactgc
      901 acctcggttc tatcgattga attcccgggc tcgagcagca tgggttgctg tttctccaag
      961 accgcagcga agggagaagc cgccgcggag aggcctgggg aggcggctgt ggcctcgtcg
     1021 ccttccaaag cgaacggaca ggagaatggc cacgtgaagg taggatcctc tagagtcgac
     1081 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     1141 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     1201 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     1261 ctcgtgacca ccttcagcta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     1321 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     1381 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     1441 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     1501 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     1561 ggcatcaagg ccaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     1621 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     1681 tacctgagcc accagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     1741 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
     1801 gcggccgcga caagcttggc cgccatggcc caacttgttt attgcagctt ataatggtta
     1861 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     1921 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatcgatc gggaattaat
     1981 tcggcgcagc accatggcct gaaataacct ctgaaagagg aacttggtta ggtaccttct
     2041 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     2101 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     2161 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     2221 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     2281 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     2341 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     2401 tgttaacagc ttggcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     2461 tacccaactt aatcgccttg cagcacatcc ccccttcgcc agctggcgta atagcgaaga
     2521 ggcccgcacc gatcgccctt cccaacagtt gcgtagcctg aatggcgaat ggcgcctgat
     2581 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca
     2641 tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     2701 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     2761 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     2821 tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt
     2881 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     2941 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcgggcta ttcttttgat
     3001 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     3061 tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca
     3121 atctgctctg atgccgcata gttaagccaa ctccgctatc gctacgtgac tgggtcatgg
     3181 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     3241 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     3301 cgtcatcacc gaaacgcgcg aggcagtatt cttgaagacg aaagggcctc gtgatacgcc
     3361 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     3421 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3481 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     3541 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     3601 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     3661 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     3721 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg
     3781 atgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     3841 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     3901 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     3961 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     4021 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgccag
     4081 cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     4141 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     4201 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     4261 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     4321 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     4381 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     4441 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4501 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4561 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4621 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4681 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     4741 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4801 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4861 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4921 gaacgaccta caccgaactg agatacctac agcgtgagca ttgagaaagc gccacgcttc
     4981 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     5041 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     5101 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     5161 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     5221 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     5281 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     5341 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatcc agctggcacg
     5401 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttacctca
     5461 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     5521 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgaa ttaa
//
