LOCUS       sequence-122342-        6120 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EGFP tagged Collybistin expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6120)
  AUTHORS   de Groot C, Floriou-Servou A, Tsai YC, Fruh S, Kohler M, Parkin G, 
            Schwerdel C, Bosshard G, Kaila K, Fritschy JM, Tyagarajan SK
  TITLE     RhoGEF9 splice isoforms influence neuronal maturation and synapse 
            formation downstream of alpha2 GABAA receptors.
  JOURNAL   PLoS Genet. 2017 Oct 25;13(10):e1007073. doi: 
            10.1371/journal.pgen.1007073. eCollection 2017 Oct.
  PUBMED    29069083
REFERENCE   2  (bases 1 to 6120)
  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"; journalName: "PLoS 
            Genet."; date: "2017-10-25"; volume: "13(10):e1007073. doi"; pages: 
            " 10.1371/journal.pgen.1007073. eCollection 2017 Oct"
COMMENT     Sequence Label: EGFPC2-CB2 SH3+
FEATURES             Location/Qualifiers
     source          1..6120
                     /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"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2908..3029
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2945..2964)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2999..3018
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3036..3491)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3173..3194)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3385..3404
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3518..3622
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3624..3981
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3624..3815
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3647..3667)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3832..3967
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4016..4810
                     /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             4016..4810
                     /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(4070..4089)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4680..4699
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4998..5017)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5042..5089
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5418..6006
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5907..5926
                     /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 gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggccggac tcagatctcg agctcaagct tcatgcagtg gattagaggc
     1381 ggatcgggaa tgctgatcac tggagattcc atcgttagtg ctgaggcagt atgggatcac
     1441 gtcaccatgg ccaaccggga gttggcattt aaggctggcg acgtcatcaa agtcttggat
     1501 gcttccaaca aggattggtg gtggggccag atcgacgatg aggagggatg gtttcctgcc
     1561 agctttgtga ggctctgggt gaaccaggag gatggggtgg aggaagggcc cagtgacgtg
     1621 cagaatgggc acctggaccc caactcagac tgcctctgtc tgggccggcc actacagaac
     1681 cgggaccaga tgcgggccaa tgtcatcaat gagatcatga gcacggaacg tcattacatc
     1741 aagcatctca aggacatttg cgaggggtac ctgaagcagt gccgaaagag aagggacatg
     1801 ttcagtgatg agcagctgaa agtcatcttt ggaaacattg aagacatcta cagatttcag
     1861 atgggcttcg tgagagactt ggagaagcag tacaacaatg atgaccccca cctcagcgag
     1921 ataggaccct gcttcttgga gcatcaagat ggcttctgga tatactctga gtattgtaac
     1981 aaccacctgg atgcctgcat ggagctctcc aaactgatga aggacagccg ctatcaacac
     2041 ttctttgagg cctgccgcct cttgcagcag atgatcgaca tcgctatcga tggttttctt
     2101 ttgactccag tacagaagat ctgcaaatac cccttacaat tggccgagct cctaaagtat
     2161 actgcccaag accacagtga ttacagatat gtcgcagctg ctttggctgt catgagaaat
     2221 gtgactcagc agatcaatga acgcaagcga cgtctggaga atattgacaa gattgctcaa
     2281 tggcaggcct ctgtcctaga ctgggaggga gatgacatct tagacaggag ctcggaactg
     2341 atctacactg gagagatggc ctggatctac cagccctatg gccgcaacca gcagagagtc
     2401 ttcttcctgt ttgaccacca gatggtcctc tgcaagaagg acctaatccg gagagacatc
     2461 ctatactaca aggggcgcat tgacatggat aaatatgagg tcattgacat tgaagatggc
     2521 agagatgatg actttaatgt cagcatgaaa aatgctttca agctgcacaa caaggaaacc
     2581 gaagaagtac atctgttctt tgccaagaaa ctggaggaga aaatacgttg gcttagagct
     2641 ttcagagaag agaggaaaat ggtacaagaa gatgaaaaaa ttggctttga gatttctgaa
     2701 aatcagaaga gacaggctgc aatgactgtg agaaaagcgt ctaaacagaa agtaacccaa
     2761 agaaaatggc actattaggg atccaccgga tctagataac tgatcataat cagccatacc
     2821 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     2881 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     2941 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     3001 ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt
     3061 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     3121 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     3181 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     3241 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     3301 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     3361 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     3421 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     3481 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3541 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     3601 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     3661 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3721 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3781 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3841 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3901 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     3961 ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat
     4021 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     4081 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     4141 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga
     4201 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     4261 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     4321 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     4381 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     4441 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     4501 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga
     4561 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     4621 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     4681 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     4741 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     4801 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     4861 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     4921 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     4981 cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     5041 acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg
     5101 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     5161 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     5221 gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag
     5281 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     5341 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     5401 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     5461 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     5521 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     5581 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     5641 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     5701 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     5761 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     5821 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     5881 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     5941 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     6001 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     6061 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
//
