LOCUS       sequence-140555-        8487 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express GFP-nArgBP2 fusion protein in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8487)
  AUTHORS   Zhang Q, Gao X, Li C, Feliciano C, Wang D, Zhou D, Mei Y, Monteiro 
            P, Anand M, Itohara S, Dong X, Fu Z, Feng G
  TITLE     Impaired Dendritic Development and Memory in Sorbs2 Knock-Out Mice.
  JOURNAL   J Neurosci. 2016 Feb 17;36(7):2247-60. doi: 
            10.1523/JNEUROSCI.2528-15.2016.
  PUBMED    26888934
REFERENCE   2  (bases 1 to 8487)
  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: 
            "10.1523/JNEUROSCI.2528-15"; journalName: "J Neurosci"; date: 
            "2016-02-17- 17"; volume: "36"; issue: "7"; pages: "2247-60"
COMMENT     Sequence Label: GFP-nArgBP2
FEATURES             Location/Qualifiers
     source          1..8487
                     /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"
     RBS             2025..2033
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             3745..3774
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             5066..5077
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     polyA_signal    5275..5396
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5312..5331)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5366..5385
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5403..5858)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5540..5561)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5752..5771
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5885..5989
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5991..6348
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5991..6182
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6014..6034)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6199..6334
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6383..7177
                     /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             6383..7177
                     /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(6437..6456)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7047..7066
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7365..7384)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7409..7456
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7785..8373
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8274..8293
                     /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 agctccatga atacagatag cggtgggtgt
     1381 gctcgcaaac gtgccgccat gtctgttacg ttaacatctg tgaagagagt tcaaagttct
     1441 ccaaacctat tggctgcagg gcgtgagtct cagtctccag attcagcaaa aggttttaga
     1501 agcgttcggc caaacctgca agacaaaaga tcaccgaccc agagccagat cactatcaat
     1561 ggcaactctg gtggcgccgt gagtccagtg agttactatc agaggccatt ctccccttct
     1621 gcatactccc tcccagcctc gctgaactcc agcattatca tgcagcacgg caggtctctt
     1681 gattctgcag agacatattc ccagcatgcc cagtccctgg atggcaccat gggaagctcc
     1741 atcccgctct atagatcctc cgaggaagag aagagggtca cggtcatcaa agccccgcat
     1801 tacccaggga tcggccctgt ggatgagtct ggaatcccca cagccattag gacgactgtc
     1861 gacaggccaa aggactggta caagacaatg ttcaagcaaa tccacatggt acacaagccg
     1921 ggtctgtaca actcgcccta cagtgctcag tcccacccgg ctgcgaagac tcagacctac
     1981 agacctcttt cgaaaagcca ctcagacaat ggcaccgatg cgtttaagga ggtgccctcc
     2041 ccagtgcctc ccccacacgt cccaccacga ccaagagatc agtcttcaac actaaagcat
     2101 gactgggacc ctccagacag aaaggtggac accaggaaat ttcgatcgga gccaaggagt
     2161 atttttgaat atgagcctgg gaagtcgtcc attctgcagc acgaaagacc cccccctctc
     2221 ccaccaactc cgacacctgt tcccagggaa cccagccgga agcctctctc agtgtcaccc
     2281 tcaacagatg gcctgcgtag cccttcccct ccgcccagga gctgtgtccc cgcaccccgt
     2341 ccgagtgccc cagatctctc tcccacccgg acgggtcgca taaatccagc tgacatagat
     2401 ttagaaaatg aaccctggta taaattcttt tcagaactgg agtttggaca cccgcctcct
     2461 aaaaaggctc tggattatgt tcaagatcat tcttccggtg tgtccaacga ggtctccatc
     2521 tatcagtcct ccatagacag aagcctggaa agacccagca gctctgcaag catggccggt
     2581 gactttagga aacggaggaa gagcgaacct gcagtgggcc cactccgggg cttgggggat
     2641 cagagttcaa gcaggaccag ccctggccgg gcggacctcc caggatcaag ttccaccttt
     2701 acgaagtctt tcattagctc ttctccttcc tctccctcga gagcacaagg tggggatgat
     2761 agcaaaatgt gtccgcccct ttgcagttac tcggggctca atggcacccc ctctggtgag
     2821 ttagagtgct gcaacgctta cagacaacat ttggacgtcc ccggggactc gcaaagggcc
     2881 atcactttca agaatggctg gcaaatggcc cggcaaaatg cagagatctg gagtagcact
     2941 gaagagaccg tttcccccaa aatcaaatca cgtagctgcg atgatctcct gaatgacgac
     3001 tgcgacagct tcccagaccc taaaaccaag tcagagagta tgggttctct gttatgtgaa
     3061 gaagactcca aagagagctg ccccatgaca tgggcgtccc cctacatcca ggaagtgtgc
     3121 gggaacagca gatctaggct caaacacagg tcagcccata atgccccagg cttcctcaaa
     3181 atgtacaaga aaatgcatcg catcaaccgc aaggacttga tgaattcaga ggtcatttgc
     3241 tcggtgaaat ccaggatcct gcagtacgag aaggagcaac agcacagggg gctgctgcac
     3301 ggatggagcc agtcttccac cgaggaggtg cccagggacg tggtgcccac tcgcatttcg
     3361 gaatttgaaa agctgattca gaagtcgaag tctatgccca atctaggaga tgaaatgtta
     3421 tctcctataa ccctagaacc cccacaaaat ggtttgtgtc ccaagaggcg attttctatc
     3481 gagtctctgc tggaggaaga aactcaggtt cgtcaccctt ctcagggtca gcgaagctgc
     3541 aagtctaaca ccctggtgcc catccacatc gaggtcacca gcgacgagca acctagaaca
     3601 cacatggagt tttccgacag tgaccaagat ggggttgtgt ccgaccacag cgactacgtt
     3661 cacgtcgaag ggtcgtcctt ttgtagtgaa agtgactttg accacttttc attcacatcc
     3721 tctgaaagct tctacggatc gagccatcac caccaccatc accatcacca tcaccgacac
     3781 ctcatcagct cctgcaaagg ccgatgcccg gcttcttaca ctcgatttac cacaatgcta
     3841 aaacatgaaa gagctaagca tgaaaacatg gaccgaccca gaaggcaaga aatggatcct
     3901 ggcctgtcta agcttgcatt tctagtcagc cctgtgcctt tccgaaggaa aaaaatcctg
     3961 actccccaaa aacagactga aaaggcaaaa tgcaaagcct cagtagttga agctctggac
     4021 tctgccctga aagacatttg cgaccaaata aaagccgaaa agcggagggg aagcttgcca
     4081 gacaacagca tcctgcacag gcttatcagt gaactgctgc cacagattcc tgaaaggaat
     4141 tcatctcttc atgctctcaa aaggagcccc atgcaccagc ctttccaccc actgcctcca
     4201 gatggtgcta gtcattgtcc cctgtaccaa aatgattgtg ggagaatgcc tcacagtgcc
     4261 tctttccctg acgtggacac aaccagcaac taccacgcac aggactatgg gagtgcactg
     4321 agcctccaag atcacgagtc ccctagaagt tactcatcta ctctgactga cttgggcaga
     4381 agtgcatcac gggaacgaag gggaactcca gaaaaagaga aattgcctgc aaaagctgtc
     4441 tatgatttca aagctcagac atctaaggag ctgtcattta agaaaggaga cactgtctac
     4501 atcctcagga aaattgacca aaactggtat gagggggaac accacggcag agtgggcatt
     4561 ttcccaatct catacgtaga gaaactaaca cccccggaaa aagcgcagcc cgcaagacca
     4621 ccacccccag tccagcccgg agagattgga gaagccatag ccaagtacaa cttcaatgca
     4681 gacacaaatg tggaactctc cctgcgaaag ggcgacagga ttattcttct taaaagagtt
     4741 gaccaaaact ggtacgaagg taaaatccca ggaaccaaca gacaaggcat cttccccgtg
     4801 tcctatgtcg aggttgtcaa gaggaacgca aaaggtgctg aggactaccc agaccctcct
     4861 ctgccccaca gctactccag tgacagaatt tacaccctaa gctccaataa gccacagcgt
     4921 cctgggttct ctcatgaaaa cattcaaggt gggggagaac cgtttcaggc tctgtataac
     4981 tatactccta ggaacgaaga tgagctggaa ctcagagaaa gtgatgtcgt tgatgtcatg
     5041 gaaaagtgtg acgacggatg gtttgtggga acttcaagaa gaaccaaatt ctttggtact
     5101 tttcccggaa actatgtcaa aaggctgtga ggtaccgcgg gcccgggatc caccggatct
     5161 agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     5221 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     5281 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     5341 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc
     5401 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     5461 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     5521 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     5581 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     5641 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     5701 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     5761 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     5821 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     5881 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     5941 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     6001 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     6061 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     6121 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     6181 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     6241 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     6301 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     6361 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     6421 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     6481 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     6541 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     6601 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     6661 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     6721 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     6781 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     6841 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     6901 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     6961 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     7021 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     7081 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     7141 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     7201 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     7261 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     7321 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     7381 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     7441 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     7501 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     7561 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     7621 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     7681 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     7741 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     7801 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     7861 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     7921 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     7981 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     8041 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     8101 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     8161 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     8221 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     8281 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     8341 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     8401 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     8461 gtggataacc gtattaccgc catgcat
//
