LOCUS       Exported                8001 bp ds-DNA     circular SYN 15-JUN-2021
DEFINITION  FRET donor for dynamically measuring TrkB activity in neurons.
ACCESSION   .
VERSION     .
KEYWORDS    TrkB mEGFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8001)
  AUTHORS   Harward SC, Hedrick NG, Hall CE, Parra-Bueno P, Milner TA, Pan E, 
            Laviv T, Hempstead BL, Yasuda R, McNamara JO
  TITLE     Autocrine BDNF-TrkB signalling within a single dendritic spine.
  JOURNAL   Nature. 2016 Sep 28;538(7623):99-103. doi: 10.1038/nature19766.
  PUBMED    27680698
REFERENCE   2  (bases 1 to 8001)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 15, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8001
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        9..388
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..666
                     /label=chicken beta-actin promoter
     primer_bind     1607..1629
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1695..1714
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      4270..4279
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             4276..4995
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             4276..4992
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4321..4342)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4582..4601)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4929..4950
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5023..5042)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5088..5143
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5142..5161)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5504..5520)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5504..5520)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5517..5539)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5528..5544
                     /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(5552..5582)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5597..5618
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5677..5872
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5723..5858
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5785..5804
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    5878..6012
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5915..5934)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5969..5988
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6080..6097)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6251..6839)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6331..6350)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7010..7870)
                     /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     7633..7652
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7871..7975)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgatcga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
       61 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      121 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      181 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      241 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      301 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      361 ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct
      421 ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt aattattttg
      481 tgcagcgatg ggggcggggg gggggggggg sgcgcgccag gcggggcggg gcggggcgag
      541 gggcggggcg gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga
      601 aagtttcctt ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg
      661 cgggcgggag tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc
      721 cgcccgcccc ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct
      781 tctcctccgg gctgtaatta gcgcttggtt taatgacggc ttgtttcttt tctgtggctg
      841 cgtgaaagcc ttgaggggct ccgggagggc cctttgtgcg gggggagcgg ctcggggggt
      901 gcgtgcgtgt gtgtgtgcgt ggggagcgcc gcgtgcggct ccgcgctgcc cggcggctgt
      961 gagcgctgcg ggcgcggcgc ggggctttgt gcgctccgca gtgtgcgcga ggggagcgcg
     1021 gccgggggcg gtgccccgcg gtgcgggggg ggctgcgagg ggaacaaagg ctgcgtgcgg
     1081 ggtgtgtgcg tgggggggtg agcagggggt gtgggcgcgt cggtcgggct gcaacccccc
     1141 ctgcaccccc ctccccgagt tgctgagcac ggcccggctt cgggtgcggg gctccgtacg
     1201 gggcgtggcg cggggctcgc cgtgccgggc ggggggtggc ggcaggtggg ggtgccgggc
     1261 ggggcggggc cgcctcgggc cggggagggc tcgggggagg ggcgcggcgg cccccggagc
     1321 gccggcggct gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag
     1381 agggcgcagg gacttccttt gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc
     1441 cgcaccccct ctagcgggcg cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc
     1501 ggggagggcc ttcgtgcgtc gccgcgccgc cgtccccttc tccctctcca gcctcggggc
     1561 tgtccgcggg gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg
     1621 cgtgtgaccg gcggctctag ctagagcctc tgctaaccat gttcatgcct tcttcttttt
     1681 cctacagctc ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt
     1741 cgctagcgct accggactca gatctcgagc tcaagcttcg aattcgccac catgtcgccc
     1801 tggctgaagt ggcatggacc cgccatggcg cggctctggg gcttatgcct gctggtcttg
     1861 ggcttctgga gggcctctct cgcctgcccg acgtcctgca aatgcagttc cgctaggatt
     1921 tggtgtactg agccttctcc aggcatcgtg gcattcccga ggttggaacc taacagcgtt
     1981 gacccggaga acatcacgga aattctcatt gcaaaccaga aaaggctaga aatcatcaat
     2041 gaagatgacg ttgaagctta cgtggggctg agaaacctta caattgtgga ttccggctta
     2101 aagtttgtgg cttacaaagc gtttctgaaa aacagcaacc tgcggcacat aaatttcaca
     2161 cgaaacaagc tgacgagttt gtccaggaga catttccgcc accttgactt gtctgacctg
     2221 atcctgacgg gtaatccgtt cacgtgctcc tgcgacatca tgtggctcaa gactctccag
     2281 gagactaaat ccagccccga cactcaggat ttgtactgcc tcaatgagag cagcaagaac
     2341 atgcccctgg cgaacctgca gatacccaat tgtggtctgc catctgcacg tctggctgct
     2401 cctaacctca ccgtggagga aggaaagtct gtgacccttt cctgcagtgt ggggggtgac
     2461 ccactcccca ccttgtactg ggacgttggg aatttggttt ccaagcacat gaatgaaaca
     2521 agccacacac agggctcctt aaggataacg aacatttcat ctgatgacag tggaaagcaa
     2581 atctcttgtg tggcagaaaa ccttgtagga gaagaccaag attctgtgaa cctcactgtg
     2641 cattttgcgc caactatcac gtttctcgag tctccaacct cagatcacca ctggtgcatt
     2701 ccattcactg tgagaggcaa ccccaagcct gcgcttcagt ggttctacaa tggggccata
     2761 ctgaatgagt ccaagtacat ctgtactaag atccacgtca ccaatcacac ggagtaccat
     2821 ggctgcctcc agctggataa ccccactcat atgaataacg gagactacac cctgatggcc
     2881 aagaacgagt atgggaagga tgagagacag atctccgctc acttcatggg ccggcctgga
     2941 gtcgactacg agacaaaccc aaattaccct gaagtcctct atgaagactg gaccacgcca
     3001 actgacattg gggatactac gaacaaaagt aatgaaatcc cctccacgga tgttgctgac
     3061 caaagcaatc gggagcatct ctcggtctat gccgtggtgg tgattgcatc tgtggtggga
     3121 ttctgcctgc tggtgatgtt gctcctgctc aagttggcga gacattccaa gtttggcatg
     3181 aaaggcccag cttcggtcat cagcaacgac gatgactctg ccagccccct ccaccacatc
     3241 tccaatggga gtaacactcc atcttcttcg gagggcggtc ccgacgctgt cattattgga
     3301 atgaccaaga ttcctgttat tgaaaacccc cagtactttg gcatcaccaa cagtcagctc
     3361 aagccagaca catttgttca gcacatcaag agacacaaca tcgttctgaa gagggaactt
     3421 ggggaaggag ccttcgggaa agttttcctt gccgagtgct acaacctctg cccagagcag
     3481 gataagatcc tggtggctgt gaagacgctg aaggacgcca gcgacaatgc acgcaaggac
     3541 tttcatcggg aagctgagct gctgaccaac ctccagcacg agcacattgt caagttctac
     3601 ggtgtctgtg tggagggcga cccactcatc atggtctttg agtacatgaa gcacggggac
     3661 ctcaacaagt tccttagggc acacgggccc gacgcagtgc tgatggcaga gggtaacccg
     3721 cccacagagc tgacgcagtc gcagatgctg cacatcgctc agcaaatcgc agcaggtatg
     3781 gtctacctgg cgtcccaaca ctttgtgcac cgtgacctgg ccacccggaa ctgcctggtg
     3841 ggagagaacc tgctggtgaa aattggggac tttgggatgt cccgagatgt gtacagcacc
     3901 gactactatc gggtcggtgg ccacacaatg ttgcccatcc gatggatgcc tccagagagc
     3961 atcatgtata ggaaattcac caccgagagc gacgtctgga gcctgggcgt tgtgttgtgg
     4021 gagatcttca cctacggcaa gcagccctgg tatcagctat cgaacaatga ggtgatagag
     4081 tgcatcaccc agggaagagt ccttcagcgg cctcgaacct gtccccagga ggtgtatgag
     4141 ctcatgctcg gatgctggca gcgggaacca cacacccgga agaacatcaa gagcatccac
     4201 accctccttc agaacttggc caaggcatct cccgtctacc tggatatcct aggcgcggat
     4261 ccaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     4321 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     4381 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     4441 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
     4501 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     4561 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     4621 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     4681 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     4741 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     4801 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     4861 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     4921 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     4981 gagctgtaca agtaaagcgg ccgcgactct agagtcgacg gatctttttc cctctgccaa
     5041 aaattatggg gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt
     5101 tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg
     5161 gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc
     5221 ccatatgctg gctgccatga acaaaggttg gctataaaga ggtcatcagt atatgaaaca
     5281 gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt
     5341 tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt
     5401 actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt
     5461 atggagatcc ctcgacctgc agcccaagct tggcgtaatc atggtcatag ctgtttcctg
     5521 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     5581 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     5641 ctttccagtc gggaaacctg tcgtgccagc ggatacgcat ctcaattagt cagcaaccat
     5701 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     5761 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     5821 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctaac
     5881 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     5941 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     6001 catgtctcga tccgctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     6061 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     6121 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     6181 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     6241 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     6301 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     6361 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     6421 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     6481 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     6541 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     6601 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     6661 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     6721 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     6781 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     6841 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     6901 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     6961 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     7021 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     7081 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     7141 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     7201 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     7261 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     7321 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     7381 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     7441 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     7501 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     7561 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     7621 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     7681 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     7741 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     7801 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     7861 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     7921 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     7981 ttccccgaaa agtgccacct g
//
