LOCUS       sequence-10214-e        7822 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 7822)
  AUTHORS   Holbro N, Grunditz A, Oertner TG
  TITLE     Differential distribution of endoplasmic reticulum controls 
            metabotropic signaling and plasticity at hippocampal synapses.
  JOURNAL   Proc Natl Acad Sci U S A. 2009 Sep 1;106(35):15055-60. doi: 
            10.1073/pnas.0905110106. Epub 2009 Aug 18.
  PUBMED    19706463
REFERENCE   2  (bases 1 to 7822)
  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: 
            "10.1073/pnas.0905110106"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2009-09-1- 1"; volume: "106"; issue: "35"; pages: "15055-60"
COMMENT     Sequence Label: pMH4-SYN-EGFP-ER
FEATURES             Location/Qualifiers
     source          1..7822
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_signal     191..341
                     /label=Ad5 Psi
                     /note="packaging signal for adenovirus serotype 5"
     promoter        393..840
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             join(925..927,928..930,931..1641)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(970..991)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1231..1250)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1578..1599
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1642..1707
                     /label=MCS
                     /note="multiple cloning site"
     CDS             5353..5544
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(5538..5560)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5646..5786
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5701..5720
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5801..5818)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5972..6560)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6052..6071)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6731..7522,7523..7591))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7354..7373
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7592..7696)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7764..7782
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcccttccag ctctctgccc cttttggatt gaagccaata tgataatgag ggggtggagt
       61 ttgtgacgtg gcgcggggcg tgggaacggg gcgggtgacg tagtagtgtg gcggaagtgt
      121 gatgttgcaa gtgtggcgga acacatgtaa gcgacggatg tggcaaaagt gacgtttttg
      181 gtgtgcgccg gtgtacacag gaagtgacaa ttttcgcgcg gttttaggcg gatgttgtag
      241 taaatttggg cgtaaccgag taagatttgg ccattttcgc gggaaaactg aataagagga
      301 agtgaaatct gaataatttt gtgttactca tagcgcgtaa tctctagcat cgaagatccc
      361 tcgactctag actgcagagg gccctgcgta tgagtgcaag tgggttttag gaccaggatg
      421 aggcggggtg ggggtgccta cctgacgacc gaccccgacc cactggacaa gcacccaacc
      481 cccattcccc aaattgcgca tcccctatca gagaggggga ggggaaacag gatgcggcga
      541 ggcgcgtgcg cactgccagc ttcagcaccg cggacagtgc cttcgccccc gcctggcggc
      601 gcgcgccacc gccgcctcag cactgaaggc gcgctgacgt cactcgccgg tcccccgcaa
      661 actccccttc ccggccacct tggtcgcgtc cgcgccgccg ccggcccagc cggaccgcac
      721 cacgcgaggc gcgagatagg ggggcacggg cgcgaccatc tgcgctgcgg cgccggcgac
      781 tcagcgctgc ctcagtctgc ggtgggcagc ggaggagtcg tgtcgtgcct gagagcgcag
      841 tcgagaattc aagctgctag catgctgcta tccgtgccgt tgctgctcgg cctcctcggc
      901 ctggccgtcg ccgaccggtc caccatggtg agcaagggcg aggagctgtt caccggggtg
      961 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     1021 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     1081 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     1141 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     1201 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     1261 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     1321 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     1381 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
     1441 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     1501 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccaagctgag caaagacccc
     1561 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     1621 ggcatggacg agctgtacaa gtccggactc agatctcgag ctcaagcttc gaattctgca
     1681 gtcgacggta ccgcgggccc gggatcctac aagaaggacg agctgtaaag atctcgagct
     1741 caagcttcga attctgcagt cgacttcgag caacttgttt attgcagctt ataatggtta
     1801 caaataaagt agcatcgaag atctgggcgt ggttaagggt gggaaagaat atataaggtg
     1861 ggggtcttat gtagttttgt atctgttttg cagcagccgc cgccgccatg agcaccaact
     1921 cgtttgatgg aagcattgtg agctcatatt tgacaacgcg catgccccca tgggccgggg
     1981 tgcgtcagaa tgtgatgggc tccagcattg atggtcgccc cgtcctgccc gcaaactcta
     2041 ctaccttgac ctacgagacc gtgtctggaa cgccgttgga gactgcagcc tccgccgccg
     2101 cttcagccgc tgcagccacc gcccgcggga ttgtgactga ctttgctttc ctgagcccgc
     2161 ttgcaagcag tgcagcttcc cgttcatccg cccgcgatga caagttgacg gctcttttgg
     2221 cacaattgga ttctttgacc cgggaactta atgtcgtttc tcagcagctg ttggatctgc
     2281 gccagcaggt ttctgccctg aaggcttcct cccctcccaa tgcggtttaa aacataaata
     2341 aaaaaccaga ctctgtttgg atttggatca agcaagtgtc ttgctgtctt tatttagggg
     2401 ttttgcgcgc gcggtaggcc cgggaccagc ggtctcggtc gttgagggtc ctgtgtattt
     2461 tttccaggac gtggtaaagg tgactctgga tgttcagata catgggcata agcccgtctc
     2521 tggggtggag gtagcaccac tgcagagctt catgctgcgg ggtggtgttg tagatgatcc
     2581 agtcgtagca ggagcgctgg gcgtggtgcc taaaaatgtc tttcagtagc aagctgattg
     2641 ccaggggcag gcccttggtg taagtgttta caaagcggtt aagctgggat gggtgcatac
     2701 gtggggatat gagatgcatc ttggactgta tttttaggtt ggctatgttc ccagccatat
     2761 ccctccgggg attcatgttg tgcagaacca ccagcacagt gtatccggtg cacttgggaa
     2821 atttgtcatg tagcttagaa ggaaatgcgt ggaagaactt ggagacgccc ttgtgacctc
     2881 caagattttc catgcattcg tccataatga tggcaatggg cccacgggcg gcggcctggg
     2941 cgaagatatt tctgggatca ctaacgtcat agttgtgttc caggatgaga tcgtcatagg
     3001 ccatttttac aaagcgcggg cggagggtgc cagactgcgg tataatggtt ccatccggcc
     3061 caggggcgta gttaccctca cagatttgca tttcccacgc tttgagttca gatgggggga
     3121 tcatgtctac ctgcggggcg atgaagaaaa cggtttccgg ggtaggggag atcagctggg
     3181 aagaaagcag gttcctgagc agctgcgact taccgcagcc ggtgggcccg taaatcacac
     3241 ctattaccgg gtgcaactgg tagttaagag agctgcagct gccgtcatcc ctgagcaggg
     3301 gggccacttc gttaagcatg tccctgactc gcatgttttc cctgaccaaa tccgccagaa
     3361 ggcgctcgcc gcccagcgat agcagttctt gcaaggaagc aaagtttttc aacggtttga
     3421 gaccgtccgc cgtaggcatg cttttgagcg tttgaccaag cagttccagg cggtcccaca
     3481 gctcggtcac ctgctctacg gcatctcgat ccagcatatc tcctcgtttc gcgggttggg
     3541 gcggctttcg ctgtacggca gtagtcggtg ctcgtccaga cgggccaggg tcatgtcttt
     3601 ccacgggcgc agggtcctcg tcagcgtagt ctgggtcacg gtgaaggggt gcgctccggg
     3661 ctgcgcgctg gccagggtgc gcttgaggct ggtcctgctg gtgctgaagc gctgccggtc
     3721 ttcgccctgc gcgtcggcca ggtagcattt gaccatggtg tcatagtcca gcccctccgc
     3781 ggcgtggccc ttggcgcgca gcttgccctt ggaggaggcg ccgcacgagg ggcagtgcag
     3841 acttttgagg gcgtagagct tgggcgcgag aaataccgat tccggggagt aggcatccgc
     3901 gccgcaggcc ccgcagacgg tctcgcattc cacgagccag gtgagctctg gccgttcggg
     3961 gtcaaaaacc aggtttcccc catgcttttt gatgcgtttc ttacctctgg tttccatgag
     4021 ccggtgtcca cgctcggtga cgaaaaggct gtccgtgtcc ccgtatacag acttgagagg
     4081 cctgtcctcg accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc
     4141 ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac
     4201 aggtgccggc agcgctctgg gtcattttcg gcgaggaccg ctttcgctgg agcgcgacga
     4261 tgatcggcct gtcgcttgcg gtattcggaa tcttgcacgc cctcgctcaa gccttcgtca
     4321 ctggtcccgc caccaaacgt ttcggcgaga agcaggccat tatcgccggc atggcggccg
     4381 acgcgctggg ctacgtcttg ctggcgttcg cgacgcgagg ctggatggcc ttccccatta
     4441 tgattcttct cgcttccggc ggcatcggga tgcccgcgtt gcaggccatg ctgtccaggc
     4501 aggtagatga cgaccatcag ggacagcttc aaggatcgct cgcggctctt accagcctaa
     4561 cttcgatcac tggaccgctg atcgtcacgg cgatttatgc cgcctcggcg agcacatgga
     4621 acgggttggc atggattgta ggcgccgccc tataccttgt ctgcctcccc gcgttgcgtc
     4681 gcggtgcatg gagccgggcc acctcgacct gaatggaagc cggcggcacc tcgctaacgg
     4741 attcaccact ccaagaattg gagccaatca attcttgcgg agaactgtga atgcgcaaac
     4801 caacccttgg cagaacatat ccatcgcgtc cgccatctcc agcagccgca cgcggcgcat
     4861 ctcgggcagc gttgggtcct ggccacgggt gcgcatgatc gtgctcctgt cgttgaggac
     4921 ccggctaggc tggcggggtt gccttactgg ttagcagaat gaatcaccga tacgcgagcg
     4981 aacgtgaagc gactgctgct gcaaaacgtc tgcgacctga gcaacaacat gaatggtctt
     5041 cggtttccgt gtttcgtaaa gtctggaaac gcggaagtca gcgccctgca ccattatgtt
     5101 ccggatctgc atcgcaggat gctgctggct accctgtgga acacctacat ctgtattaac
     5161 gaagcgctgg cattgaccct gagtgatttt tctctggtcc cgccgcatcc ataccgccag
     5221 ttgtttaccc tcacaacgtt ccagtaaccg ggcatgttca tcatcagtaa cccgtatcgt
     5281 gagcatcctc tctcgtttca tcggtatcat tacccccatg aacagaaatt cccccttaca
     5341 cggaggcatc aagtgaccaa acaggaaaaa accgccctta acatggcccg ctttatcaga
     5401 agccagacat taacgcttct ggagaaactc aacgagctgg acgcggatga acaggcagac
     5461 atctgtgaat cgcttcacga ccacgctgat gagctttacc gcagctgcct cgcgcgtttc
     5521 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     5581 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     5641 cggggcgcag ccatgaccca gtcacgtagc gatagcggag tgtatactgg cttaactatg
     5701 cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat
     5761 gcgtaaggag aaaataccgc atcaggcgct cttccgcttc ctcgctcact gactcgctgc
     5821 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     5881 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     5941 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     6001 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     6061 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6121 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta
     6181 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6241 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6301 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6361 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     6421 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6481 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     6541 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6601 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6661 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6721 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6781 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6841 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     6901 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6961 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     7021 gtttgcgcaa cgttgttgcc attgctgcag gcatcgtggt gtcacgctcg tcgtttggta
     7081 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     7141 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     7201 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     7261 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     7321 gaccgagttg ctcttgcccg gcgtcaacac gggataatac cgcgccacat agcagaactt
     7381 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     7441 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     7501 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     7561 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     7621 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     7681 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     7741 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt cttcaagaat
     7801 tgatccgggc ccccatttcc cc
//
