LOCUS       sequence-415333-        6122 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Flag tagged mammalian expression of GC domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6122)
  AUTHORS   Tyagarajan SK, Ghosh H, Yevenes GE, Imanishi SY, Zeilhofer HU, 
            Gerrits B, Fritschy JM
  TITLE     Extracellular signal-regulated kinase and glycogen synthase kinase 
            3beta regulate gephyrin postsynaptic aggregation and GABAergic 
            synaptic function in a calpain-dependent mechanism.
  JOURNAL   J Biol Chem. 2013 Apr 5;288(14):9634-47. doi: 
            10.1074/jbc.M112.442616. Epub 2013 Feb 13.
  PUBMED    23408424
REFERENCE   2  (bases 1 to 6122)
  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: 
            "10.1074/jbc.M112.442616"; journalName: "J Biol Chem"; date: 
            "2013-04-5- 5"; volume: "288"; issue: "14"; pages: "9634-47"
COMMENT     Sequence Label: pCR3-FLAG-Gephyrin GC
FEATURES             Location/Qualifiers
     source          1..6122
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        10..389
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..593
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     544..564
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     638..657
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        638..656
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             673..744
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     promoter        complement(1832..1850)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1833..1850)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1870..1887)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1876..2100
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2232..2820)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2312..2331)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3149..3196
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3221..3240
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3428..4222)
                     /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             complement(3428..4222)
                     /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(3539..3558)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4149..4168
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4257..4614)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4271..4406
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4423..4614
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4571..4591
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(4674..5465,5466..5534))
                     /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     5297..5316
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5536..5638)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5665..6120
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5752..5771)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5962..5983
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gcgcgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
       61 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      121 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      181 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      241 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      301 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      361 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      421 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      481 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      541 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc
      601 taactagaga acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga
      661 cccaagctta tggactacaa ggacgacgac gacaaggact acaaggacga cgacgacaag
      721 gactacaagg acgacgacga caagggagca ggagcgaccg agggaatgat cctcacaaac
      781 cacgaccatc aaatccgtgt cggagtcctc acagtgagtg atagctgctt caggaatctt
      841 gcagaagacc gcagtgggat aaatctaaaa gatctcgtcc aggatccttc tttgttgggt
      901 gggactatat cagcatacaa gatagtacca gatgaaatag aagaaatcaa ggaaacactc
      961 atagactggt gtgatgagaa ggaacttaat ttgatattaa caactggcgg aacagggttt
     1021 gcaccacgag atgtcactcc agaggccaca aaagaagtaa tagaacggga agcgccaggg
     1081 atggccctgg caatgctgat gggatcactt aatgttacac ctctgggcat gctctctaga
     1141 ccagtgtgtg gaataagagg gaaaactctg ataattaacc tgcctggtag caagaaagga
     1201 tctcaggaat gctttcagtt catactgcca gctctacctc atgccattga ccttttacgt
     1261 gatgccattg taaaagtaaa ggaggtgcat gatgaacttg aagatttacc ttccccacca
     1321 cctcctcttt ctccacctcc tacaactagc ccacataagc agacagaaga caaaggagtt
     1381 cagtgtgaag aggaggaaga agaaaagaaa gacagtggcg tagcttcgac agaagatagt
     1441 tcctcatcac atattactgc agcagctctt gctgcaaaga ttccagactc catcatttct
     1501 cgtggcgttc aggtgctccc acgagacaca gcctccctta gcactactcc ttcagagtcg
     1561 ccccgtgccc aggctacatc tcgcctctct acagcttctt gtccaacacc aaaagtccag
     1621 tccaggtgca gcagcaagga gaacattcta agagccagtc acagtgctgt agatatcacc
     1681 aaggtggcta gaagacatcg catgtctcct tttcctctga cgtcttgagg taccgagctc
     1741 ggatccacta gtaacggccg ccagtgtgct ggaattctgc agatatccat cacactggcg
     1801 gccgctcgag catgcatcta gagggcccta ttctatagtg tcacctaaat gctagagctc
     1861 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     1921 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     1981 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     2041 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     2101 cttctgaggc ggaaagaacc agtggcggta atacggttat ccacagaatc aggggataac
     2161 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     2221 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     2281 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     2341 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     2401 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     2461 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     2521 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     2581 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     2641 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     2701 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     2761 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     2821 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     2881 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     2941 tgaagtttta aatcaatcta aagtatatat gagtaacctg aggctatggc agggcctgcc
     3001 gccccgacgt tggctgcgag ccctgggcct tcacccgaac ttggggggtg gggtggggaa
     3061 aaggaagaaa cgcgggcgta ttggccccaa tggggtctcg gtggggtatc gacagagtgc
     3121 cagccctggg accgaacccc gcgtttatga acaaacgacc caacaccgtg cgttttattc
     3181 tgtcttttta ttgccgtcat agcgcgggtt ccttccggta ttgtctcctt ccgtgtttca
     3241 gttagcctcc ccctagggtg ggcgaagaac tccagcatga gatccccgcg ctggaggatc
     3301 atccagccgg cgtcccggaa aacgattccg aagcccaacc tttcatagaa ggcggcggtg
     3361 gaatcgaaat ctcgtgatgg caggttgggc gtcgcttggt cggtcatttc gaaccccaga
     3421 gtcccgctca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag
     3481 cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc tcttcagcaa
     3541 tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc cggccacagt
     3601 cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag gcatcgccat
     3661 gggtcacgac gagatcctcg ccgtcgggca tgctcgcctt gagcctggcg aacagttcgg
     3721 ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga ccggcttcca
     3781 tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg
     3841 gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc tcggcaggag
     3901 caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc cagtcccttc
     3961 ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg gccagccacg
     4021 atagccgcgc tgcctcgtct tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa
     4081 aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag cagccgattg
     4141 tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga gaacctgcgt
     4201 gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga tcgatctttg
     4261 caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc agaggccgag
     4321 gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg agaatgggcg
     4381 gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac
     4441 taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac
     4501 ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg
     4561 gactttccac accctaactg acacacattc cacagctggt tctttccgcc tcaggactct
     4621 tcctttttca ataaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     4681 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     4741 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     4801 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     4861 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     4921 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     4981 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     5041 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     5101 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     5161 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     5221 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     5281 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     5341 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     5401 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     5461 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     5521 tgttgaatac tcatatcttc ctttttcaat attattgaag catttatcag ggttattgtc
     5581 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     5641 catttccccg aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     5701 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     5761 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     5821 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     5881 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     5941 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     6001 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     6061 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     6121 ac
//
