LOCUS       sequence-417807-        6908 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mCherry tagged Geph for mammalian expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6908)
  AUTHORS   Tyagarajan SK, Ghosh H, Yevenes GE, Nikonenko I, Ebeling C, 
            Schwerdel C, Sidler C, Zeilhofer HU, Gerrits B, Muller D, Fritschy 
            JM
  TITLE     Regulation of GABAergic synapse formation and plasticity by 
            GSK3beta-dependent phosphorylation of gephyrin.
  JOURNAL   Proc Natl Acad Sci U S A. 2011 Jan 4;108(1):379-84. doi: 
            10.1073/pnas.1011824108. Epub 2010 Dec 20.
  PUBMED    21173228
REFERENCE   2  (bases 1 to 6908)
  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.1011824108"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2011-01-4- 4"; volume: "108"; issue: "1"; pages: "379-84"
COMMENT     Sequence Label: pmCherryC2-Gephyrin P1
FEATURES             Location/Qualifiers
     source          1..6908
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             597..1304
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(742..760)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1010..1029
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3696..3817
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3733..3752)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3787..3806
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3824..4279)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3961..3982)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4173..4192
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4306..4410
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4412..4769
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4412..4603
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4435..4455)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4620..4755
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4804..5598
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4804..5598
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4858..4877)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5468..5487
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5786..5805)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5830..5877
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6206..6794
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6695..6714
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcatgg
      601 tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc
      661 acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
      721 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg
      781 cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg
      841 ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga
      901 tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg
      961 agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc
     1021 agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc
     1081 tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg
     1141 tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca
     1201 tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg
     1261 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagggcgct atctcgagct
     1321 caagcttcga attcatggcg accgagggaa tgatcctcac aaaccacgac catcaaatcc
     1381 gtgtcggagt cctcacagtg agtgatagct gcttcaggaa tcttgcagaa gaccgcagtg
     1441 ggataaatct aaaagatctc gtccaggatc cttctttgtt gggtgggact atatcagcat
     1501 acaagatagt accagatgaa atagaagaaa tcaaggaaac actcatagac tggtgtgatg
     1561 agaaggaact taatttgata ttaacaactg gcggaacagg gtttgcacca cgagatgtca
     1621 ctccagaggc cacaaaagaa gtaatagaac gggaagcgcc agggatggcc ctggcaatgc
     1681 tgatgggatc acttaatgtt acacctctgg gcatgctctc tagaccagtg tgtggaataa
     1741 gagggaaaac tctgataatt aacctgcctg gtagcaagaa aggatctcag gaatgctttc
     1801 agttcatact gccagctcta cctcatgcca ttgacctttt acgtgatgcc attgtaaaag
     1861 taaaggaggt gcatgatgaa cttgaagatt taccttcccc accacctcct ctttctccac
     1921 ctcctacaac tagcccacat aagcagacag aagacaaagg agttcagtgt gaagaggagg
     1981 aagaagaaaa gaaagacagt ggcgtagctt cgacagaaga tagttcctca tcacatatta
     2041 ctgcagcagc tcttgctgca aagattccag actccatcat ttctcgtggc gttcaggtgc
     2101 tcccacgaga cacagcctcc cttagcacta ctccttcaga gtcgccccgt gcccaggcta
     2161 catctcgcct ctctacagct tcttgtccaa caccaaaagt ccagtccagg tgcagcagca
     2221 aggagaacat tctaagagcc agtcacagtg ctgtagatat caccaaggtg gctagaagac
     2281 atcgcatgtc tccttttcct ctgacgtcta tggacaaagc cttcattaca gttctggaga
     2341 tgactccggt gcttggtaca gaaatcatca attaccgaga tggaatgggg cgagtccttg
     2401 ctcaagatgt gtatgcaaaa gacaacctac ccccattccc agcatcagta aaagatggct
     2461 atgctgttcg agctgctgat ggtccaggag atcgtttcat cattggggaa tcccaagctg
     2521 gtgaacagcc aactcagaca gtaatgcccg gacaagtgat gcgggttaca acaggtgctc
     2581 caatcccctg cggtgctgat gcagtagtgc aagttgaaga tactgaactc atcagggaat
     2641 cagatgatgg tactgaagaa cttgaagtgc gcattctggt gcaagctcgg ccaggccaag
     2701 acatcagacc catcggccac gacattaaaa gaggggagtg tgttttggca aaaggaaccc
     2761 atatgggccc ttcagaaatt ggtctccttg caactgtagg tgtcacagaa gttgaagtta
     2821 ataagtttcc agtggttgcc gttatgtcaa cagggaatga gctactaaat cctgaagatg
     2881 acctcttacc aggaaagatt cgggacagca atcgatcaac acttctagca acaattcagg
     2941 aacatggtta cccaacaatc aacttgggaa ttgtaggaga caacccagat gacttactca
     3001 atgccttgaa tgagggtatc agccgtgcta atgtcatcat cacgtcagga ggtgtatcca
     3061 tgggggaaaa ggactatctc aagcaggtgc tggacatcga tcttcatgct cagatccatt
     3121 ttggcagggt ttttatgaag ccaggcttgc caacaacatt tgcaactttg gatattgatg
     3181 gtgtaagaaa aattattttt gcactaccag ggaatcctgt atcagctgtg gtcacctgca
     3241 acctctttgt tgtacccgca ctgagaaaaa tgcagggtat cttggatcct cggccaacca
     3301 tcatcaaagc taggttatca tgtgacgtaa agctggaccc tcgcccagaa taccaccggt
     3361 gtatactgac ttggcatcac caagaaccac tgccctgggc ccagagtaca ggtaatcaga
     3421 tgagcagccg tctgatgagc atgcgcagtg ccaatggatt gctgatgcta cctccaaaga
     3481 cagagcagta tgtggaactt cacaaaggcg aggtggtgga cgtcatggtc atcggacggc
     3541 tatgactcga cggtaccgcg ggcccgggat ccaccggatc tagataactg atcataatca
     3601 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     3661 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     3721 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     3781 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat
     3841 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     3901 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     3961 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     4021 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     4081 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     4141 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     4201 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     4261 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     4321 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     4381 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     4441 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4501 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4561 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     4621 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     4681 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     4741 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     4801 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     4861 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     4921 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     4981 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     5041 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     5101 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     5161 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     5221 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     5281 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     5341 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     5401 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     5461 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     5521 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     5581 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     5641 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     5701 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     5761 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     5821 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     5881 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     5941 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     6001 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     6061 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     6121 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     6181 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     6241 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     6301 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     6361 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     6421 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     6481 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     6541 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     6601 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     6661 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     6721 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     6781 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     6841 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     6901 ccatgcat
//
