LOCUS       sequence-65200-e        5867 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 5867)
  AUTHORS   Gross GG, Junge JA, Mora RJ, Kwon HB, Olson CA, Takahashi TT, Liman 
            ER, Ellis-Davies GC, McGee AW, Sabatini BL, Roberts RW, Arnold DB
  TITLE     Recombinant probes for visualizing endogenous synaptic proteins in 
            living neurons.
  JOURNAL   Neuron. 2013 Jun 19;78(6):971-85. doi: 10.1016/j.neuron.2013.04.017.
  PUBMED    23791193
REFERENCE   2  (bases 1 to 5867)
  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.2013.04"; journalName: "Neuron"; date: 
            "2013-06-19- 19"; volume: "78"; issue: "6"; pages: "971-85"
COMMENT     Sequence Label: pGW_GPHN.FingR-YFP
FEATURES             Location/Qualifiers
     source          1..5867
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             283..306
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(325..327,328..1041)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(367..388)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(628..647)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     975..996
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1157..1291
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1194..1213)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1248..1267
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1315..1331)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1315..1331)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1328..1350)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1339..1355
                     /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(1363..1369,1370..1387,1388..1393))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1408..1429
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1546..1563)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1717..2305)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1797..1816)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2476..3267,3268..3336))
                     /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     3099..3118
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     3546..3564
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3602..3624)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3724..3743
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3975..3997
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3989..4006
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3990..4006
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        4556..4935
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4936..5139
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5090..5110
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5136..5160
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 aagcttatta caatgctcga ggtcaaggaa gcatcaccaa ccagcatcca gatcagctgg
       61 ggcaagtaca aggtcatggt tcgctactac cgcatcacct acggtgaaac tggtggcaat
      121 agccctgtcc aggaattcac cgtgcctggc agcaagtcca ctgctaccat cagcagcctg
      181 aaacctggtg tcgactatac catcacggtg tacgccgtca cgatcgacca ctggaactac
      241 caggacccga tcccgatctc catcaactac cgcaccggat ccgattacaa ggatgacgac
      301 gataagggta gcggctcctc tagagtgagc aagggcgagg agctgttcac cggggtggtg
      361 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
      421 ggcgagggcg atgccaccta cggcaagctg accctgaagc tgatctgcac caccggcaag
      481 ctgcccgtgc cctggcccac cctcgtgacc accctgggct acggcctgca gtgcttcgcc
      541 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
      601 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
      661 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
      721 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
      781 accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag
      841 gacggcggcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
      901 gtgctgctgc ccgacaacca ctacctgagc taccagtccg ccctgagcaa agaccccaac
      961 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     1021 atggacgagc tgtacaagta ggaattcatt gatcataatc agccatacca catttgtaga
     1081 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     1141 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     1201 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     1261 actcatcaat gtatcttatc atgtctggat ccgtaccgag ctcgcgtaat catggtcata
     1321 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     1381 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     1441 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     1501 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     1561 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     1621 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     1681 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     1741 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     1801 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     1861 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg
     1921 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     1981 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2041 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2101 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     2161 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     2221 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     2281 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     2341 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     2401 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     2461 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     2521 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     2581 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     2641 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     2701 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     2761 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     2821 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     2881 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     2941 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     3001 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     3061 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     3121 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     3181 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     3241 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     3301 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aataagcggc
     3361 cgcggccatg ccggccacta gtctcgagtt attattgaag catttatcag ggttattgtc
     3421 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     3481 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     3541 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     3601 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     3661 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     3721 atgcggcatc agagcagatt gtactgagag tgcaccatat gaagacgtcg cctcctcact
     3781 acttctggaa tagctcagag gccgaggcgg cctcggcctc tgcataaata aaaaaaatta
     3841 gtcagcgcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     3901 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     3961 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca tctgcagtga
     4021 ataataaaat gtgtgtttgt ccgaaatacg cgtttgagat ttctgtcccg actaaattca
     4081 tgtcgcgcga tagtggtgtt tatcgccgat agagatggcg atattggaaa aatcgatatt
     4141 tgaaaatatg gcatattgaa aatgtcgccg atgtgagttt ctgtgtaact gatatcgcca
     4201 tttttccaaa agttgatttt tgggcatacg cgatatctgg cgatacgctt atatcgttta
     4261 cgggggatgg cgatagacgc ctttggtgac ttgggcgatt ctgtgtgtcg caaataatcg
     4321 cagtttcgat ataggtgaca gacgatatga ggctatatcg ccgatagagg cgacatcaag
     4381 ctggcacatg gccaatgcat atcgatctat acattgaatc aatattggcc attagccata
     4441 ttattcattg gttatatagc ataaatcaat attggctatt ggccattgca tacgttgtat
     4501 ccatatcata atatgtacat ttatattggc tcatgtccaa cattaccgcc atgttgacat
     4561 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
     4621 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     4681 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     4741 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     4801 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     4861 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
     4921 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
     4981 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
     5041 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
     5101 ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcagatc
     5161 gcctggagac gccatccacg ctgttttgac ctccatagaa gacaccggga ccgatccagc
     5221 ctccgcggcc gggaacggtg cattggaacg cggattcccc gtgccaagag tgacgtaagt
     5281 accgcctata gagtctatag gcccaccccc ttggcttctt atgcatgcta tactgttttt
     5341 ggcttggggt ctatacaccc ccgcttcctc atgttatagg tgatggtata gcttagccta
     5401 taggtgtggg ttattgacca ttattgacca ctcccctatt ggtgacgata ctttccatta
     5461 ctaatccata acatggctct ttgcactgtc cttcagagac tgacacggac tctgtatttt
     5521 tacaggatgg ggtctcattt attatcaact ctctttattg gctatatgcc aatacattac
     5581 aaattcacat atacaacacc accgtcccca gtgcccgcag tttttattaa acataacgtg
     5641 ggatctccag cgaatctcgg gtacgtgttc cggacatggg gctcttctcc ggtagcggcg
     5701 gagtttctac atccgagccc tgctcccatg cctccagcga ctcatggtcg ctcggcagct
     5761 ccttgctcct aacagtggag gccagactta ggcacagcac gatgcccacc accaccagtg
     5821 tgccgcacaa ggccgtggcg gtagggtatg tgtctgaaaa tgagctc
//
