LOCUS       sequence-181352-        6339 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FRET probe; extracellular membrane tethered eGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6339)
  AUTHORS   Octeau JC, Chai H, Jiang R, Bonanno SL, Martin KC, Khakh BS
  TITLE     An Optical Neuron-Astrocyte Proximity Assay at Synaptic Distance 
            Scales.
  JOURNAL   Neuron. 2018 Apr 4;98(1):49-66.e9. doi: 
            10.1016/j.neuron.2018.03.003.
  PUBMED    29621490
REFERENCE   2  (bases 1 to 6339)
  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: 
            "10.1016/j.neuron.2018.03"; journalName: "Neuron"; date: "2018-04-4-
            4"; volume: "98"; issue: "1"; pages: "49-66.e9"
COMMENT     Sequence Label: pcDNA3.1 CMV NAPA-A BGH
FEATURES             Location/Qualifiers
     source          1..6339
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        60..439
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        440..643
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     594..614
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     688..707
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        688..706
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     sig_peptide     747..806
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             join(810..812,813..1523)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(810..812,813..1523)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(852..873)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1113..1132)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1460..1481
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1530..1559
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1563..1709
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     primer_bind     complement(1755..1772)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1761..1985
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2031..2459
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2118..2137)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2328..2349
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2468..2488)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2473..2802
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2653..2788
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2715..2734
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2869..3663
                     /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(2923..2942)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3533..3552
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3837..3958
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3874..3893)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3928..3947
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4007..4023)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4007..4023)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4020..4042)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4031..4047
                     /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(4055..4061,4062..4079,4080..4085))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4100..4121
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4238..4255)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4409..4997)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4489..4508)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5168..5959,5960..6028))
                     /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     5791..5810
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6029..6133)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6208..6227)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg
       61 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
      121 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
      181 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      241 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      301 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      361 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      421 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      481 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      541 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      601 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga
      661 acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg
      721 ctagcgttta aacttaagct accaccatgg agacagacac actcctgcta tgggtactgc
      781 tgctctgggt tccaggttcc actggtgacg tgagcaaggg cgaggagctg ttcaccgggg
      841 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
      901 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
      961 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
     1021 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     1081 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     1141 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     1201 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     1261 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
     1321 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     1381 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
     1441 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     1501 tcggcatgga cgagctgtac aagaagcttg aacaaaaact catctcagaa gaggatctga
     1561 atgctgtggg ccaggacacg caggaggtca tcgtggtgcc acactccttg ccctttaagg
     1621 tggtggtgat ctcagccatc ctggccctgg tggtgctcac catcatctcc cttatcatcc
     1681 tcatcatgct ttggcagaag aagccacgtt agggccgctc gagtctagag ggcccgttta
     1741 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     1801 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     1861 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     1921 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     1981 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     2041 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     2101 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     2161 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     2221 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     2281 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     2341 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     2401 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     2461 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     2521 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     2581 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     2641 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     2701 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     2761 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     2821 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     2881 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     2941 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     3001 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     3061 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     3121 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     3181 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     3241 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     3301 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     3361 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     3421 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     3481 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     3541 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     3601 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     3661 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     3721 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     3781 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     3841 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     3901 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     3961 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     4021 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     4081 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     4141 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     4201 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     4261 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     4321 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4381 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4441 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4501 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4561 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4621 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4681 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4741 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4801 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     4861 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4921 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4981 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5041 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5101 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5161 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5221 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5281 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5341 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5401 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5461 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5521 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     5581 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     5641 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     5701 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5761 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     5821 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     5881 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5941 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6001 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6061 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6121 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     6181 tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     6241 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
     6301 tacaacaagg caaggcttga ccgacaattg catgaagaa
//
