LOCUS       sequence-301951-        7683 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 7683)
  AUTHORS   Kang MG, Guo Y, Huganir RL
  TITLE     AMPA receptor and GEF-H1/Lfc complex regulates dendritic spine 
            development through RhoA signaling cascade.
  JOURNAL   Proc Natl Acad Sci U S A. 2009 Mar 3. 106(9):3549-54.
  PUBMED    19208802
REFERENCE   2  (bases 1 to 7683)
  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: "Proc Natl 
            Acad Sci U S A. 2009 Mar 3. 106(9):3549-54."
COMMENT     Sequence Label: GEF-DN
FEATURES             Location/Qualifiers
     source          1..7683
                     /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    4471..4592
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4508..4527)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4562..4581
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4599..5054)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4736..4757)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4948..4967
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5081..5185
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5187..5544
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5187..5378
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5210..5230)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5395..5530
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5579..6373
                     /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             5579..6373
                     /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(5633..5652)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6243..6262
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6561..6580)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6605..6652
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6981..7569
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7470..7489
                     /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 actcagatct cgagctcaag cttcgaattc tgcagtcgac catgtctgga
     1381 tcgaatccct cactcgcgcg cggatcgacc ggagcaagga gcaggcgacc aagacccggg
     1441 aaaaggagaa gatgaaggaa gctaaagatg cccgctatac caacggccac ctcttcacca
     1501 ccatctcggt ctccggcatg actatgtgct atgcctgtaa caagagcatt acagccaagg
     1561 aagccctcat ttgtcccaca tgtaacgtga ccatccacaa ccgctgtaag gacacgctgg
     1621 ccaactgtac caaggtcaag cagaagcaac agaaagctgc actgctgagg aacaacactg
     1681 ccttgcagtc cgtctctctc cgaagtaaga caaccaccag agagcggcca acgtctgcca
     1741 tctacccttc cgatagcttc cggcagtccc tcctgggttc tcgtcgtggc ctgtcctcct
     1801 tgtctttggc caaaagtgtt tccactacca acattgctgg acatttcaat gatgagtctc
     1861 ctctggggct gcgtcagatc ctctcccagt ccacagactc cctcaacatg cggaaccgaa
     1921 ccctgtccgt ggagtccctt attgatgaag gtgtagaagt gttctacaac gagctcatga
     1981 gtgactttga gatggacgag aaggacttcg aggcggattc atggagcctt gctgtggaca
     2041 gcagcttcct gcaacagcat aaaaaggagg tgatgaagaa gcaagatgtc atctacgagc
     2101 tgatccaggc cgagctgcac catgtgagaa ccttgaagat tatgacccgc ctcttccgca
     2161 ctgggatgct ggaagagttg cagatggagc cagaagtggt ccagggactg ttcccctgtg
     2221 tggacgagct tagtgacatt cacacacgtt tccttagtca gcttttggaa cggcggcgcc
     2281 aggccctgtg tccaggcagc acccggaact ttgtcatcca tcgtttgggt gacttgctca
     2341 tcagccagtt ctcaggttcc aacgctgagc agatgcgcaa gacttactca gagttctgca
     2401 gccgccacac caaggcctta aagctctata aggagctgta cgctcgagac aaacgcttcc
     2461 aacagttcat ccggaaaatg acccgctctg ctgtgctgaa gcggcatggg gttcaggagt
     2521 gcattctcct ggtaactcag cggattacca aatacccagt gctcatcaac aggatcctgc
     2581 agaattccca tgggattgaa gaggagtacc aagacttggc agcagcccta gggctagtaa
     2641 aggagttgtt gtccaatgtg gaccaggatg tacacgagct ggagaaagag gcccgccttc
     2701 aggagattta caaccgaatg gatcctcggg ctcagacccc tgttcctggc aagggcccct
     2761 tcggccgaga tgaacttttg cggagaaaac ttatccatga tggctgcctg ctctggaaga
     2821 cagccacagg ccgctttaaa gatgtcctct tactactgat gacagatgtg ctggtgtttc
     2881 tccaggaaaa ggaccagaag tacattttca cgtccctgga caagccctca gtggtatcct
     2941 tgcagaatct catcgtaaga gacattgcca accaggcgaa agggatgttt ctgatcagtt
     3001 ctggcccacc tgaaatgtat gaagtacacg cagcatctcg agatgaccgt actacctgga
     3061 tccgtgtcat ccagcagagc gtgcgcttat gcccgtcgag ggaggacttc cctctgatcg
     3121 agacagagga taaggcgtac ctccggagga tcaagacgaa actgcagcag aaaaaccagg
     3181 cgctggtgga gctgctacag atgaatgttg agctatttgc tgagatggtc cacttccagg
     3241 cattaaaagc tggtttcatc ggaatgcccc cgcccaccct gcccaggggc cttttccgtc
     3301 ttgagtcctt tgagtccctt cgaggcgagc gtctgctcaa agatgccctc cgtgaagtgg
     3361 aaggcctgaa agacctgttg ttgggccctt gtgtggacct gcctttgaca gcccgagaac
     3421 cagccttacc cgtagaagct gacagtggta gctgtcctgg ggtcactgcc aacggagagg
     3481 ccagaacctt caatggctcc attgagctct gtagagcaga ctcagattcc agccagaagg
     3541 atcggaatgg aaatcagttg aggtcaccac aggaggaggc gttacaacca ttggtcaatc
     3601 tgtatggact cctacaaggc ctgcaggctg ttgtggtcca gcaagaaaga ttgatggaag
     3661 ccctgttccc tgaaggccct gagcggtggg aaaagctatc ccgagccaac tctcgggatg
     3721 gcgaagctgg ccgggctgcg gttgcttctg taactcccga gaagcaggcc acggagctgg
     3781 cattactgca gaggcagcac agcctgttgc aggaagagct gcggcgctgc cagcggctcg
     3841 gggaagagcg ggcaactgaa gctggcagcc tggaggccag gctgcgggag agcgagcaag
     3901 cccgggccct gctggagcgg gaggctgaag agatccgccg gcagctagca gccttgggcc
     3961 aaaacgagcc actcccggca gaagcaccct gggctcgcag gcctctggac cctcggcgcc
     4021 gcagccttcc agcaggcgac gctctgtact tgagcttcaa tcccccccag cctagtcgag
     4081 gccatgaccg cctggatttg cctgtgactg ttcgttccct ccaccgaccc tttgatgacc
     4141 gagaggcaca agaacttggt agccccgagg atcgactgca ggacagcagt gaccctgata
     4201 cttgcagtga ggaggaagtc agtagccggc tgtccccacc ccacagtcca cgagacttca
     4261 cccgaatgca ggacattccc gaagagacag aaagccgaga tggggagcct acggcctcag
     4321 agagctaagg ccgcgggccc gggatccacc ggatctagat aactgatcat aatcagccat
     4381 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     4441 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     4501 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     4561 tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg ttaatatttt
     4621 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     4681 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     4741 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     4801 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     4861 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     4921 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     4981 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     5041 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     5101 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     5161 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
     5221 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     5281 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     5341 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     5401 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     5461 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     5521 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
     5581 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     5641 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     5701 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     5761 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     5821 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     5881 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     5941 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     6001 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     6061 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     6121 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     6181 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     6241 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     6301 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     6361 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     6421 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     6481 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     6541 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     6601 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
     6661 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     6721 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     6781 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
     6841 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     6901 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     6961 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     7021 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     7081 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     7141 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     7201 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     7261 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     7321 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     7381 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     7441 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     7501 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     7561 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     7621 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg
     7681 cat
//
