LOCUS       sequence-7785-ex        8154 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 8154)
  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 8154)
  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-C
FEATURES             Location/Qualifiers
     source          1..8154
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        398..701
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        702..905
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     856..876
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      964..973
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(970..972,973..975,976..1686)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(970..972,973..975,976..1686)
                     /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(1015..1036)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1276..1295)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1623..1644
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1910..2031
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1947..1966)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2001..2020
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5376..5393)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5547..6135)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5627..5646)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             6946..7740
                     /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             6946..7740
                     /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(7000..7019)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7610..7629
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7850..7871)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8062..8081
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tcccttccag ctctctgccc cttttggatt gaagccaata tgataatgag ggggtggagt
       61 ttgtgacgtg gcgcggggcg tgggaacggg gcgggtgacg tagtagtgtg gcggaagtgt
      121 gatgttgcaa gtgtggcgga acacatgtaa gcgacggatg tggcaaaagt gacgtttttg
      181 gtgtgcgccg gtgtacacag gaagtgacaa ttttcgcgcg gttttaggcg gatgttgtag
      241 taaatttggg cgtaaccgag taagatttgg ccattttcgc gggaaaactg aataagagga
      301 agtgaaatct gaataatttt gtgttactca tagcgcgtaa nnnntaatag taatcaatta
      361 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      421 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      481 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      541 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      601 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      661 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      721 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      781 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      841 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      901 gagctggttt agtgaaccgt cagatccgct agagatcgat ctggtaccgt cgacgtcgcg
      961 gccgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     1021 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     1081 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     1141 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     1201 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     1261 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     1321 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     1381 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     1441 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     1501 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     1561 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     1621 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     1681 tacaagtccg gactcagatc tgtaggtgca ttaatggaag tgatgcagaa gaggagcaaa
     1741 gccattcatt cttcagactg aagcttctag ataagatatc cgatccaccg gatctagata
     1801 actgatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     1861 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     1921 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     1981 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aacgcggatc
     2041 tgggcgtggt taagggtggg aaagaatata taaggtgggg gtcttatgta gttttgtatc
     2101 tgttttgcag cagccgccgc cgccatgagc accaactcgt ttgatggaag cattgtgagc
     2161 tcatatttga caacgcgcat gcccccatgg gccggggtgc gtcagaatgt gatgggctcc
     2221 agcattgatg gtcgccccgt cctgcccgca aactctacta ccttgaccta cgagaccgtg
     2281 tctggaacgc cgttggagac tgcagcctcc gccgccgctt cagccgctgc agccaccgcc
     2341 cgcgggattg tgactgactt tgctttcctg agcccgcttg caagcagtgc agcttcccgt
     2401 tcatccgccc gcgatgacaa gttgacggct cttttggcac aattggattc tttgacccgg
     2461 gaacttaatg tcgtttctca gcagctgttg gatctgcgcc agcaggtttc tgccctgaag
     2521 gcttcctccc ctcccaatgc ggtttaaaac ataaataaaa aaccagactc tgtttggatt
     2581 tggatcaagc aagtgtcttg ctgtctttat ttaggggttt tgcgcgcgcg gtaggcccgg
     2641 gaccagcggt ctcggtcgtt gagggtcctg tgtatttttt ccaggacgtg gtaaaggtga
     2701 ctctggatgt tcagatacat gggcataagc ccgtctctgg ggtggaggta gcaccactgc
     2761 agagcttcat gctgcggggt ggtgttgtag atgatccagt cgtagcagga gcgctgggcg
     2821 tggtgcctaa aaatgtcttt cagtagcaag ctgattgcca ggggcaggcc cttggtgtaa
     2881 gtgtttacaa agcggttaag ctgggatggg tgcatacgtg gggatatgag atgcatcttg
     2941 gactgtattt ttaggttggc tatgttccca gccatatccc tccggggatt catgttgtgc
     3001 agaaccacca gcacagtgta tccggtgcac ttgggaaatt tgtcatgtag cttagaagga
     3061 aatgcgtgga agaacttgga gacgcccttg tgacctccaa gattttccat gcattcgtcc
     3121 ataatgatgg caatgggccc acgggcggcg gcctgggcga agatatttct gggatcacta
     3181 acgtcatagt tgtgttccag gatgagatcg tcataggcca tttttacaaa gcgcgggcgg
     3241 agggtgccag actgcggtat aatggttcca tccggcccag gggcgtagtt accctcacag
     3301 atttgcattt cccacgcttt gagttcagat ggggggatca tgtctacctg cggggcgatg
     3361 aagaaaacgg tttccggggt aggggagatc agctgggaag aaagcaggtt cctgagcagc
     3421 tgcgacttac cgcagccggt gggcccgtaa atcacaccta ttaccgggtg caactggtag
     3481 ttaagagagc tgcagctgcc gtcatccctg agcagggggg ccacttcgtt aagcatgtcc
     3541 ctgactcgca tgttttccct gaccaaatcc gccagaaggc gctcgccgcc cagcgatagc
     3601 agttcttgca aggaagcaaa gtttttcaac ggtttgagac cgtccgccgt aggcatgctt
     3661 ttgagcgttt gaccaagcag ttccaggcgg tcccacagct cggtcacctg ctctacggca
     3721 tctcgatcca gcatatctcc tcgtttcgcg ggttggggcg gctttcgctg tacggcagta
     3781 gtcggtgctc gtccagacgg gccagggtca tgtctttcca cgggcgcagg gtcctcgtca
     3841 gcgtagtctg ggtcacggtg aaggggtgcg ctccgggctg cgcgctggcc agggtgcgct
     3901 tgaggctggt cctgctggtg ctgaagcgct gccggtcttc gccctgcgcg tcggccaggt
     3961 agcatttgac catggtgtca tagtccagcc cctccgcggc gtggcccttg gcgcgcagct
     4021 tgcccttgga ggaggcgccg cacgaggggc agtgcagact tttgagggcg tagagcttgg
     4081 gcgcgagaaa taccgattcc ggggagtagg catccgcgcc gcaggccccg cagacggtct
     4141 cgcattccac gagccaggtg agctctggcc gttcggggtc aaaaaccagg tttcccccat
     4201 gctttttgat gcgtttctta cctctggttt ccatgagccg gtgtccacgc tcggtgacga
     4261 aaaggctgtc cgtgtccccg tatacagact nnngtttaaa cgaattcnnn atataaaatg
     4321 caaggtgctg ctcaaaaaat caggcaaagc ctcgcgcaaa aaagaaagca catcgtagtc
     4381 atgctcatgc agataaaggc aggtaagctc cggaaccacc acagaaaaag acaccatttt
     4441 tctctcaaac atgtctgcgg gtttctgcat aaacacaaaa taaaataaca aaaaaacatt
     4501 taaacattag aagcctgtct tacaacagga aaaacaaccc ttataagcat aagacggact
     4561 acggccatgc cggcgtgacc gtaaaaaaac tggtcaccgt gattaaaaag caccaccgac
     4621 agctcctcgg tcatgtccgg agtcataatg taagactcgg taaacacatc aggttgattc
     4681 atcggtcagt gctaaaaagc gaccgaaata gcccggggga atacataccc gcaggcgtag
     4741 agacaacatt acagccccca taggaggtat aacaaaatta ataggagaga aaaacacata
     4801 aacacctgaa aaaccctcct gcctaggcaa aatagcaccc tcccgctcca gaacaacata
     4861 cagcgcttca cagcggcagc ctaacagtca gccttaccag taaaaaagaa aacctattaa
     4921 aaaaacacca ctcgacacgg caccagctca atcagtcaca gtgtaaaaaa gggccaagtg
     4981 cagagcgagt atatatagga ctaaaaaatg acgtaacggt taaagtccac aaaaaacacc
     5041 cagaaaaccg cacgcgaacc tacgcccaga aacgaaagcc aaaaaaccca caacttcctc
     5101 aaatcgtcac ttccgttttc ccacgttacg taacttccca ttttaagaaa actacaattc
     5161 ccaacacata caagttactc cgccctaaaa cctacgtcac ccgccccgtt cccacgcccc
     5221 gcgccacgtc acaaactcca ccccctcatt atcatattgg cttcaatcca aaataaggta
     5281 tattattgat gatnnnttaa ttaaggatcc nnncggtgtg aaataccgca cagatgcgta
     5341 aggagaaaat accgcatcag gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     5401 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     5461 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     5521 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5581 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5641 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5701 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     5761 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     5821 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     5881 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     5941 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     6001 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     6061 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     6121 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     6181 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     6241 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     6301 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     6361 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     6421 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     6481 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     6541 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6601 cgcaacgttg ttgnnnnaaa aaggatcttc acctagatcc ttttcacgta gaaagccagt
     6661 ccgcagaaac ggtgctgacc ccggatgaat gtcagctact gggctatctg gacaagggaa
     6721 aacgcaagcg caaagagaaa gcaggtagct tgcagtgggc ttacatggcg atagctagac
     6781 tgggcggttt tatggacagc aagcgaaccg gaattgccag ctggggcgcc ctctggtaag
     6841 gttgggaagc cctgcaaagt aaactggatg gctttctcgc cgccaaggat ctgatggcgc
     6901 aggggatcaa gctctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     6961 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     7021 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     7081 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     7141 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     7201 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     7261 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     7321 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     7381 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     7441 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     7501 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     7561 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     7621 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     7681 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     7741 attttgttaa aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     7801 catcccttat aaatcaaaag aatagaccgc gatagggttg agtgttgttc cagtttggaa
     7861 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     7921 gggcgatggc ccactacgtg aaccatcacc caaatcaagt tttttgcggt cgaggtgccg
     7981 taaagctcta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     8041 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     8101 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcg cgcttaatgc gccg
//
