LOCUS       sequence-156678-        7889 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpression vector for Venus-MDGA2 in neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7889)
  AUTHORS   Loh KH, Stawski PS, Draycott AS, Udeshi ND, Lehrman EK, Wilton DK, 
            Svinkina T, Deerinck TJ, Ellisman MH, Stevens B, Carr SA, Ting AY
  TITLE     Proteomic Analysis of Unbounded Cellular Compartments: Synaptic 
            Clefts.
  JOURNAL   Cell. 2016 Aug 25;166(5):1295-1307.e21. doi: 
            10.1016/j.cell.2016.07.041.
  PUBMED    27565350
REFERENCE   2  (bases 1 to 7889)
  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.cell.2016.07"; journalName: "Cell"; date: "2016-08-25- 
            25"; volume: "166"; issue: "5"; pages: "1295-1307.e21"
COMMENT     Sequence Label: pCAG-Venus-MDGA2
FEATURES             Location/Qualifiers
     source          1..7889
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1670
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1594..1616
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1678..1697
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1726..1735
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1840..1842,1843..1845,1846..2556)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1885..1906)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2146..2165)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2493..2514
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5392..5408)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5392..5408)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5405..5427)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5416..5432
                     /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(5440..5446,5447..5464,5465..5470))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5485..5506
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5565..5760
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5611..5746
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5766..5900
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5803..5822)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5857..5876
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5968..5985)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6139..6727)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6219..6238)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6898..7689,7690..7758))
                     /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     7521..7540
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7759..7863)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
     1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
     1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
     1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
     1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     1681 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attccgccac catggatttc
     1741 ctgctgctcg gtctctgtct atactggctg ctgaggaggc cctcgggggt ggtcttgtgt
     1801 ctgctggggg cctgctttca gatgctgccc gccgctagca tggtgagcaa gggcgaggag
     1861 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     1921 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagctg
     1981 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgggctac
     2041 ggcctgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     2101 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     2161 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     2221 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     2281 agccacaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc caacttcaag
     2341 atccgccaca acatcgagga cggcggcgtg cagctcgccg accactacca gcagaacacc
     2401 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccgcc
     2461 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     2521 gccgggatca ctctcggcat ggacgagctg tacaagggct cgggctcggg caccggtccc
     2581 ccgacggttc ggattgtgca ctcaggattg gcctgtaata tagaggagga gcgctactcg
     2641 gaaagggtct ataccatccg ggaaggagaa acgctagagt tgacctgtct ggttactgga
     2701 catccacgcc cacagatcag gtggaccaaa acagcaggaa gcgcctctga cagattccaa
     2761 gactcaagtg tcttcaatga gactttgagg attacaagca ttcagagaca ccaaggaggc
     2821 cgatattact gtaaagcaga gaatggcttg gggtctccag caataaagtc gatccgagtg
     2881 gatgtgtact acttggatga tcctgtagta actgttcatc aaagtatagg tgaagctaaa
     2941 gaacaatttt actatgagag aacagtgttc ctccgatgtg tggccaattc caacccccct
     3001 gtgcgctaca gctggagacg tggccaagag gtgttgctac aaggatcaga caaaggagtg
     3061 gagatttacg agcccttttt tacccagggg gaaacaaaga tcttaaagct aaagaatctc
     3121 cgacctcagg actatgctaa ctacagctgc attgcttcag tgaggaacgt gtgtaatatt
     3181 ccagacaaga tggtgtcttt tagactttcc aataaaaccg cttctccgtc aattaagctc
     3241 ttggtggatg accctatagt tgtaaatcct ggagaggcca taacgttagt gtgtgttaca
     3301 acagggggag agcctatgcc ctctctcacc tgggtcaggt ccttcgggac tctgcctgaa
     3361 aagattgttt tgaatggagg gacattgacc atacctgcca tcacctctga tgatgctggg
     3421 acttacagct gcattgccaa taataatgtg ggaaaccctg caaaaaagtc caccaacatc
     3481 attgtgagag cattaaaaaa aggacgattt tggatcactc ctgatcctta tcacaaagat
     3541 gacaacatcc agattgggcg tgaggtgaaa atatcttgcc aagtagaagc cgttccctct
     3601 gaagagttaa cattcagttg gtttaaaaac ggccgtccat taagaagttc tgagcggatg
     3661 gttattacac agactgaccc tgatgtgtcc cccggaacaa caaatttgga tatcattgat
     3721 ttaaaattca cggactttgg gacttacaca tgcgtggcct ctctgaaggg aggaggaata
     3781 tctgatatca gtattgacgt taacatttcc agcagcacag ttccacccaa tctgactgtt
     3841 ccacaggaaa aatcaccttt ggtcaccaga gaaggggaca caatagaact tcagtgtcaa
     3901 gtaactggga aaccaaaacc aatcatcctt tggtctagag cagataaaga agttgcaatg
     3961 cctgatggga caatgcaaat ggagagttat gatgggaccc tcaggattgt gaacgtgtca
     4021 agggaaatgt ctgggatgta cagatgccag accagtcagt acaatggatt caacgtgaag
     4081 cccagagaag ccttggtgca gctcatcgtg cagtatcccc cagctgtgga accagcattc
     4141 ttggaaatcc gtcaaggaca agacagaagt gtgaccatga gctgtagggt tctaagagcc
     4201 tatccgatac gtgtgctgac ttacgaatgg cgtctgggca ataaattatt acggaccggt
     4261 cagtttgact ctcaggagta tactgagtac ccactaaaga gtctttccaa tgaaaactat
     4321 ggagtttata actgcagcat cattaatgaa gctggagcag ggagatgcag ctttcttgtg
     4381 acaggaaaag cctatgcccc agagttctac tatgacacct acaaccctgt gtggcagaac
     4441 cgacaccgag tgtattctta cagtctgcag tggacacaaa tgaatcccga tgcagtggat
     4501 cggattgtcg cataccgttt gggtattcgg caggctggac agcaacgttg gtgggaacag
     4561 gagattaaaa taaatgggaa tattcaaaag ggagaactaa ttacatataa cttgactgaa
     4621 cttataaaac cagaggccta tgaagtccga ctgactcctc tcactaaatt tggtgaagga
     4681 gattcaacaa ttcgtgtaat taaatatacg gcacctgtaa atcctcattt gagagaattt
     4741 cactgtggat ttgaagatgg taatatatgc ttattcaccc aagatgatac agataacttt
     4801 gactggacaa agcaaagtac tgcaacaaga aatacaaaat acacccctaa cacaggtccc
     4861 aatgcagatc ggagtggttc caaagaaggt ttttatatgt acatcgagac atcgcgaccc
     4921 agactagaag gagaaaaggc tcgacttctc agccctgtat tcagcatagc tcccaaaaat
     4981 ccatatggac ctaccaatag tgcatattgt ttcagtttct tctatcacat gtatgggcaa
     5041 catataggtg ttttaaatgt atatctacgt ttgaaagggc agacgacaat agagaatcct
     5101 ctatggtctt cgagtgggaa caaaggacaa cgatggaatg aagctcatgt taatatatat
     5161 ccaattactt catttcagtt aatttttgaa ggcattcggg gtcctgggat agaaggtgac
     5221 atcgccattg acgatgtgtc aattgcagaa ggagaatgtg caaaacaaga cctaccaact
     5281 aagaattccg tggatggtgc tgttgggatc ttagttcata tatggctttt tccagttatc
     5341 atcctcatct cgatcttaag ccctcgaagg tgaaagcttg gcgtaatcat ggtcatagct
     5401 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     5461 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     5521 actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg atccgcatct caattagtca
     5581 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     5641 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     5701 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     5761 aagctaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     5821 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     5881 tatcttatca tgtctggatc cgctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5941 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     6001 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     6061 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     6121 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     6181 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     6241 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     6301 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc
     6361 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6421 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6481 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6541 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     6601 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6661 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6721 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6781 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6841 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6901 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6961 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     7021 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     7081 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     7141 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     7201 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     7261 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     7321 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     7381 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7441 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7501 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7561 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7621 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7681 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7741 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7801 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7861 gcgcacattt ccccgaaaag tgccacctg
//
