LOCUS       sequence-156675-        7951 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpression vector for Venus-MDGA1 in neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7951)
  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 7951)
  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-MDGA1
FEATURES             Location/Qualifiers
     source          1..7951
                     /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(5454..5470)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5454..5470)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5467..5489)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5478..5494
                     /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(5502..5508,5509..5526,5527..5532))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5547..5568
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5627..5822
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5673..5808
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5828..5962
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5865..5884)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5919..5938
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6030..6047)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6201..6789)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6281..6300)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6960..7751,7752..7820))
                     /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     7583..7602
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7821..7925)
                     /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 caccggtcac
     2581 tgccggggac aaggagtcta cgctccagcc caggcgcaga tcgtgcatgc gggccaggca
     2641 tgtgtggtga aagaggacaa tatcagcgag cgtgtctaca ccatccggga gggggacacc
     2701 ctcatgctgc agtgccttgt aacagggcac cctcgacccc aggtacggtg gaccaagacg
     2761 gcaggtagcg cctcggacaa gttccaggag acatcggtgt tcaacgagac gctgcgcatc
     2821 gagcgtattg cacgcacgca gggcggccgc tactactgca aggctgagaa cggcgtgggg
     2881 gtgccggcca tcaagtccat ccgcgtggac gtgcagtacc tggatgagcc aatgctgacg
     2941 gtgcaccaga cggtgagcga tgtgcgaggc aacttctacc aggagaagac ggtgttcctg
     3001 cgctgtactg tcaactccaa cccgcctgcc cgcttcatct ggaagcgggg ttccgatacc
     3061 ctatcccaca gccaggacaa tggggttgac atctatgagc ccctctacac tcagggggag
     3121 accaaggtcc tgaagctgaa gaacctgcgg ccccaggact atgccagcta cacctgccag
     3181 gtgtctgtgc gtaacgtgtg cggcatccca gacaaggcca tcaccttccg gctcaccaac
     3241 accacggcac caccagccct gaagctgtct gtgaacgaaa ctctggtggt gaaccctggg
     3301 gagaatgtga cggtgcagtg tctgctgaca ggcggtgatc ccctccccca gctgcagtgg
     3361 tcccatgggc ctggcccact gcccctgggt gctctggccc agggtggcac cctcagcatc
     3421 ccttcagtgc aggcccggga ctctggctac tacaactgca cagccaccaa caatgtgggc
     3481 aaccctgcca agaagactgt caacctgctg gtgcgatcca tgaagaacgc tacattccag
     3541 atcactcctg acgtgatcaa agagagtgag aacatccagc tgggccagga cctgaagcta
     3601 tcgtgccacg tggatgcagt gccccaggag aaggtgacct accagtggtt caagaatggc
     3661 aagccggcac gcatgtccaa gcggctgctg gtgacccgca atgatcctga gctgcccgca
     3721 gtcaccagca gcctagagct cattgacctg cacttcagtg actatggcac ctacctgtgc
     3781 atggcttctt tcccaggggc acccgtgccc gacctcagcg tcgaggtcaa catctcctct
     3841 gagacagtgc cgcccaccat cagtgtgccc aagggtaggg ccgtggtgac cgtgcgcgag
     3901 ggatcgcctg ccgagctgca atgcgaggtg cggggcaagc cgcggccgcc agtgctctgg
     3961 tcccgcgtgg acaaggaggc tgcactgctg ccctcggggc tgcccctgga ggagactccg
     4021 gacgggaagc tgcggctgga gcgagtgagc cgagacatga gcgggaccta ccgctgccag
     4081 acggcccgct ataatggctt caacgtgcgc ccccgtgagg cccaggtgca gctgaacgtg
     4141 cagttcccgc cggaggtgga gcccagttcc caggacgtgc gccaggcgct gggccggccc
     4201 gtgctcctgc gctgctcgct gctgcgaggc agcccccagc gcatcgcctc ggctgtgtgg
     4261 cgtttcaaag ggcagctgct gccgccgccg cctgttgttc ccgccgccgc cgaggcgccg
     4321 gatcacgcgg agctgcgcct cgacgccgta actcgcgaca gcagcggcag ctacgagtgc
     4381 agcgtctcca acgatgtggg ctcggctgcc tgcctcttcc aggtctccgc caaagcctac
     4441 agcccggagt tttacttcga cacccccaac cccacccgca gccacaagct gtccaagaac
     4501 tactcctacg tgctgcagtg gactcagagg gagcccgacg ctgtcgaccc tgtgctcaac
     4561 tacagactca gcatccgcca gttgaaccag cacaatgcgg tggtcaaggc catcccggtc
     4621 cggcgtgtgg agaaggggca gctgctggag tacatcctga ccgatctccg tgtgccccac
     4681 agctatgagg tccgcctcac accctatacc accttcgggg ctggtgacat ggcctcccgc
     4741 atcatccact acacagagcc catcaactct ccgaaccttt cagacaacac ctgccacttt
     4801 gaggatgaga agatctgtgg ctatacccag gacctgacag acaactttga ctggacgcgg
     4861 cagaatgccc tcacccagaa ccccaaacgc tcccccaaca ctggtccccc caccgacata
     4921 agtggcaccc ctgagggcta ctacatgttc atcgagacat cgaggcctcg ggagctgggg
     4981 gaccgtgcaa ggttagtgag tcccctctac aatgccagcg ccaagttcta ctgtgtctcc
     5041 ttcttctacc acatgtacgg gaaacacatc ggctccctca acctcctggt gcggtcccgg
     5101 aacaaagggg ctctggacac gcacgcctgg tctctcagtg gcaataaggg caatgtgtgg
     5161 cagcaggccc atgtgcccat cagccccagt gggcccttcc agattatttt tgagggggtt
     5221 cgaggcccgg gctacctggg ggatattgcc atagatgacg tcacactgaa gaagggggag
     5281 tgtccccgga agcagacgga tcccaataaa gtggtggtga tgccgggcag tggagccccc
     5341 tgccagtcca gcccacagct gtgggggccc atggccatct tcctcttggc gttgcagaga
     5401 tgaggcgcgc ccctgcaggg aattcgatat caagcaagct tggcgtaatc atggtcatag
     5461 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     5521 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     5581 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc ggatccgcat ctcaattagt
     5641 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     5701 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     5761 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     5821 aaaagctaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     5881 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     5941 tgtatcttat catgtctgga tccgctgcat taatgaatcg gccaacgcgc ggggagaggc
     6001 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     6061 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     6121 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     6181 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     6241 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     6301 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     6361 gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt
     6421 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     6481 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     6541 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     6601 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     6661 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     6721 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     6781 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     6841 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     6901 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     6961 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     7021 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     7081 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     7141 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     7201 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     7261 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     7321 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     7381 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     7441 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     7501 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     7561 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     7621 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     7681 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     7741 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     7801 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     7861 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     7921 ccgcgcacat ttccccgaaa agtgccacct g
//
