LOCUS       sequence-7784-ex        8212 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 8212)
  AUTHORS   Strasser GA, Rahim NA, VanderWaal KE, Gertler FB, Lanier LM
  TITLE     Arp2/3 is a negative regulator of growth cone translocation.
  JOURNAL   Neuron. 2004 Jul 8. 43(1):81-94.
  PUBMED    15233919
REFERENCE   2  (bases 1 to 8212)
  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: "Neuron. 
            2004 Jul 8. 43(1):81-94."
COMMENT     Sequence Label: pShuttle-CMV-EGFP-CA
FEATURES             Location/Qualifiers
     source          1..8212
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        409..712
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        713..916
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     867..887
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      975..984
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(981..983,984..986,987..1697)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(981..983,984..986,987..1697)
                     /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(1026..1047)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1287..1306)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1634..1655
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1979..2100
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2016..2035)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2070..2089
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5434..5451)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5605..6193)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5685..5704)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             7004..7798
                     /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             7004..7798
                     /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(7058..7077)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7668..7687
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7908..7929)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8120..8139
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ttaattaann ntcccttcca gctctctgcc ccttttggat tgaagccaat atgataatga
       61 gggggtggag tttgtgacgt ggcgcggggc gtgggaacgg ggcgggtgac gtagtagtgt
      121 ggcggaagtg tgatgttgca agtgtggcgg aacacatgta agcgacggat gtggcaaaag
      181 tgacgttttt ggtgtgcgcc ggtgtacaca ggaagtgaca attttcgcgc ggttttaggc
      241 ggatgttgta gtaaatttgg gcgtaaccga gtaagatttg gccattttcg cgggaaaact
      301 gaataagagg aagtgaaatc tgaataattt tgtgttactc atagcgcgta annnntaata
      361 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
      421 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      481 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
      541 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
      601 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
      661 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
      721 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
      781 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
      841 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
      901 ctatataagc agagctggtt tagtgaaccg tcagatccgc tagagatcga tctggtaccg
      961 tcgacgtcgc ggccgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc
     1021 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     1081 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     1141 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     1201 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     1261 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     1321 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     1381 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     1441 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     1501 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     1561 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     1621 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     1681 tggacgagct gtacaagtcc ggactcagat ctgtaggtgc attaatggaa gtgatgcaga
     1741 agaggagcaa agccattcat tcttcagacg aagatgagga tgaagatgat gatgaagatt
     1801 ttgaggatga tgatgaatgg gaagactgaa agcttctaga taagatatcc gatccaccgg
     1861 atctagataa ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     1921 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     1981 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     2041 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     2101 acgcnnnnta agggtgggaa agaatatata aggtgggggt cttatgtagt tttgtatctg
     2161 ttttgcagca gccgccgccg ccatgagcac caactcgttt gatggaagca ttgtgagctc
     2221 atatttgaca acgcgcatgc ccccatgggc cggggtgcgt cagaatgtga tgggctccag
     2281 cattgatggt cgccccgtcc tgcccgcaaa ctctactacc ttgacctacg agaccgtgtc
     2341 tggaacgccg ttggagactg cagcctccgc cgccgcttca gccgctgcag ccaccgcccg
     2401 cgggattgtg actgactttg ctttcctgag cccgcttgca agcagtgcag cttcccgttc
     2461 atccgcccgc gatgacaagt tgacggctct tttggcacaa ttggattctt tgacccggga
     2521 acttaatgtc gtttctcagc agctgttgga tctgcgccag caggtttctg ccctgaaggc
     2581 ttcctcccct cccaatgcgg tttaaaacat aaataaaaaa ccagactctg tttggatttg
     2641 gatcaagcaa gtgtcttgct gtctttattt aggggttttg cgcgcgcggt aggcccggga
     2701 ccagcggtct cggtcgttga gggtcctgtg tattttttcc aggacgtggt aaaggtgact
     2761 ctggatgttc agatacatgg gcataagccc gtctctgggg tggaggtagc accactgcag
     2821 agcttcatgc tgcggggtgg tgttgtagat gatccagtcg tagcaggagc gctgggcgtg
     2881 gtgcctaaaa atgtctttca gtagcaagct gattgccagg ggcaggccct tggtgtaagt
     2941 gtttacaaag cggttaagct gggatgggtg catacgtggg gatatgagat gcatcttgga
     3001 ctgtattttt aggttggcta tgttcccagc catatccctc cggggattca tgttgtgcag
     3061 aaccaccagc acagtgtatc cggtgcactt gggaaatttg tcatgtagct tagaaggaaa
     3121 tgcgtggaag aacttggaga cgcccttgtg acctccaaga ttttccatgc attcgtccat
     3181 aatgatggca atgggcccac gggcggcggc ctgggcgaag atatttctgg gatcactaac
     3241 gtcatagttg tgttccagga tgagatcgtc ataggccatt tttacaaagc gcgggcggag
     3301 ggtgccagac tgcggtataa tggttccatc cggcccaggg gcgtagttac cctcacagat
     3361 ttgcatttcc cacgctttga gttcagatgg ggggatcatg tctacctgcg gggcgatgaa
     3421 gaaaacggtt tccggggtag gggagatcag ctgggaagaa agcaggttcc tgagcagctg
     3481 cgacttaccg cagccggtgg gcccgtaaat cacacctatt accgggtgca actggtagtt
     3541 aagagagctg cagctgccgt catccctgag caggggggcc acttcgttaa gcatgtccct
     3601 gactcgcatg ttttccctga ccaaatccgc cagaaggcgc tcgccgccca gcgatagcag
     3661 ttcttgcaag gaagcaaagt ttttcaacgg tttgagaccg tccgccgtag gcatgctttt
     3721 gagcgtttga ccaagcagtt ccaggcggtc ccacagctcg gtcacctgct ctacggcatc
     3781 tcgatccagc atatctcctc gtttcgcggg ttggggcggc tttcgctgta cggcagtagt
     3841 cggtgctcgt ccagacgggc cagggtcatg tctttccacg ggcgcagggt cctcgtcagc
     3901 gtagtctggg tcacggtgaa ggggtgcgct ccgggctgcg cgctggccag ggtgcgcttg
     3961 aggctggtcc tgctggtgct gaagcgctgc cggtcttcgc cctgcgcgtc ggccaggtag
     4021 catttgacca tggtgtcata gtccagcccc tccgcggcgt ggcccttggc gcgcagcttg
     4081 cccttggagg aggcgccgca cgaggggcag tgcagacttt tgagggcgta gagcttgggc
     4141 gcgagaaata ccgattccgg ggagtaggca tccgcgccgc aggccccgca gacggtctcg
     4201 cattccacga gccaggtgag ctctggccgt tcggggtcaa aaaccaggtt tcccccatgc
     4261 tttttgatgc gtttcttacc tctggtttcc atgagccggt gtccacgctc ggtgacgaaa
     4321 aggctgtccg tgtccccgta tacagactnn ngtttaaacg aattcnnnat ataaaatgca
     4381 aggtgctgct caaaaaatca ggcaaagcct cgcgcaaaaa agaaagcaca tcgtagtcat
     4441 gctcatgcag ataaaggcag gtaagctccg gaaccaccac agaaaaagac accatttttc
     4501 tctcaaacat gtctgcgggt ttctgcataa acacaaaata aaataacaaa aaaacattta
     4561 aacattagaa gcctgtctta caacaggaaa aacaaccctt ataagcataa gacggactac
     4621 ggccatgccg gcgtgaccgt aaaaaaactg gtcaccgtga ttaaaaagca ccaccgacag
     4681 ctcctcggtc atgtccggag tcataatgta agactcggta aacacatcag gttgattcat
     4741 cggtcagtgc taaaaagcga ccgaaatagc ccgggggaat acatacccgc aggcgtagag
     4801 acaacattac agcccccata ggaggtataa caaaattaat aggagagaaa aacacataaa
     4861 cacctgaaaa accctcctgc ctaggcaaaa tagcaccctc ccgctccaga acaacataca
     4921 gcgcttcaca gcggcagcct aacagtcagc cttaccagta aaaaagaaaa cctattaaaa
     4981 aaacaccact cgacacggca ccagctcaat cagtcacagt gtaaaaaagg gccaagtgca
     5041 gagcgagtat atataggact aaaaaatgac gtaacggtta aagtccacaa aaaacaccca
     5101 gaaaaccgca cgcgaaccta cgcccagaaa cgaaagccaa aaaacccaca acttcctcaa
     5161 atcgtcactt ccgttttccc acgttacgta acttcccatt ttaagaaaac tacaattccc
     5221 aacacataca agttactccg ccctaaaacc tacgtcaccc gccccgttcc cacgccccgc
     5281 gccacgtcac aaactccacc ccctcattat catattggct tcaatccaaa ataaggtata
     5341 ttattgatga tnnnttaatt aaggatccnn ncggtgtgaa ataccgcaca gatgcgtaag
     5401 gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     5461 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     5521 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5581 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5641 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5701 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5761 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     5821 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5881 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5941 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6001 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     6061 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6121 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     6181 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6241 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     6301 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     6361 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     6421 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     6481 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     6541 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     6601 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     6661 caacgttgtt gnnnnaaaaa ggatcttcac ctagatcctt ttcacgtaga aagccagtcc
     6721 gcagaaacgg tgctgacccc ggatgaatgt cagctactgg gctatctgga caagggaaaa
     6781 cgcaagcgca aagagaaagc aggtagcttg cagtgggctt acatggcgat agctagactg
     6841 ggcggtttta tggacagcaa gcgaaccgga attgccagct ggggcgccct ctggtaaggt
     6901 tgggaagccc tgcaaagtaa actggatggc tttctcgccg ccaaggatct gatggcgcag
     6961 gggatcaagc tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     7021 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     7081 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     7141 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg
     7201 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     7261 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     7321 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     7381 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     7441 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     7501 gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt
     7561 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     7621 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     7681 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     7741 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgaat
     7801 tttgttaaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaca
     7861 tcccttataa atcaaaagaa tagaccgcga tagggttgag tgttgttcca gtttggaaca
     7921 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     7981 gcgatggccc actacgtgaa ccatcaccca aatcaagttt tttgcggtcg aggtgccgta
     8041 aagctctaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     8101 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     8161 gtgtagcggt cacgctgcgc gtaaccacca cacccgcgcg cttaatgcgc cg
//
