LOCUS       sequence-129577-        5433 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 5433)
  AUTHORS   Mao T, Kusefoglu D, Hooks BM, Huber D, Petreanu L, Svoboda K
  TITLE     Long-Range Neuronal Circuits Underlying the Interaction between 
            Sensory and Motor Cortex.
  JOURNAL   Neuron. 2011 Oct 6;72(1):111-23.
  PUBMED    21982373
REFERENCE   2  (bases 1 to 5433)
  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."; 
            date: "2011-10-6"; volume: "72(1)"; pages: "111-23"
COMMENT     Sequence Label: AAV-CAG-GFP
FEATURES             Location/Qualifiers
     source          1..5433
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   433..562
                     /label=AAV2 ITR
     rep_origin      634..1089
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(721..740)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     931..952
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1115..1219
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1220..1288,1289..2080)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1438..1457)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2251..2839
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2740..2759
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   3076..3205
                     /label=AAV2 ITR
     polyA_signal    3367..3488
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3404..3423)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3458..3477
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    3531..4119
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3637..3648
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     4046..4066
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(4160..4873,4874..4876,4877..4879))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(4163..4873,4874..4876,4877..4879))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4205..4226)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4554..4573
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4813..4834
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4876..4885
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(4931..4950)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(5012..5034)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
ORIGIN
        1 cccccccccc ccccccgccc ccatcgctgc acaaaataat taaaaaataa ataaatacaa
       61 aattgggggt ggggaggggg gggagatggg gagagtgaag cagaacgtgg ggctcacctc
      121 gaccatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa agtcccataa
      181 ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt caataggggg
      241 cgtacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc gtaaatactc
      301 cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat acgtcattat
      361 tgacgtcaat ggtactcttc ctagagcatg gctacgtaga taagtagcat ggcgggttaa
      421 tcattaacta caaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
      481 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
      541 cagtgagcga gcgagcgcgc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
      601 cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg
      661 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
      721 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
      781 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
      841 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
      901 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
      961 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     1021 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     1081 ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     1141 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     1201 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     1261 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     1321 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     1381 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     1441 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     1501 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     1561 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     1621 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     1681 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     1741 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     1801 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     1861 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     1921 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     1981 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     2041 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     2101 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     2161 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     2221 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     2281 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     2341 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     2401 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     2461 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     2521 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     2581 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     2641 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     2701 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     2761 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     2821 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     2881 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     2941 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     3001 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     3061 gattcattaa tgcagctgcg cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg
     3121 cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg
     3181 gccaactcca tcactagggg ttccttgtag ttaatgatta acccgccatg ctacttatct
     3241 acgtagccat gctctaggaa gatccttatc gattttacca catttgtaga ggttttactt
     3301 gctttaaaaa acctcccaca tctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     3361 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     3421 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     3481 gtatcttatc atgtctgctc gaagcggccg cccgggtcga cggtatcgat gcggggaggc
     3541 ggcccaaagg gagatccgac tcgtctgagg gcgaaggcga agacgcggaa gaggccgcag
     3601 agccggcagc aggccgcggg aaggaaggtc cgctggattg agggccgaag ggacgtagca
     3661 gaaggacgtc ccgcgcagaa tccaggtggc aacacaggcg agcagccaag gaaaggacga
     3721 tgatttcccc gacaacacca cggaattgtc agtgcccaac agccgagccc ctgtccagca
     3781 gcgggcaagg caggcggcga tgagttccgc cgtggcaata gggaggggga aagcgaaagt
     3841 cccggaaagg agctgacagg tggtggcaat gccccaacca gtgggggttg cgtcagcaaa
     3901 cacagtgcac accacgccac gttgcctgac aacgggccac aactcctcat aaagagacag
     3961 caaccaggat ttatacaagg aggagaaaat gaaagccata cgggaagcaa tagcatgata
     4021 caaaggcatt aaagcagcgt atccacatag cgtaaaagga gcaacatagt taagaatacc
     4081 agtcaatctt tcacaaattt tgtaatccag aggttgatta tcgataagct tgatatcgaa
     4141 ttctagagtc gcggccgctt tacttgtaca gctcgtccat gccgagagtg atcccggcgg
     4201 cggtcacgaa ctccagcagg accatgtgat cgcgcttctc gttggggtct ttgctcaggg
     4261 cggactgggt gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg
     4321 tgttctgctg gtagtggtcg gcgagctgca cgctgccgtc ctcgatgttg tggcggatct
     4381 tgaagttcac cttgatgccg ttcttctgct tgtcggccat gatatagacg ttgtggctgt
     4441 tgtagttgta ctccagcttg tgccccagga tgttgccgtc ctccttgaag tcgatgccct
     4501 tcagctcgat gcggttcacc agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt
     4561 agttgccgtc gtccttgaag aagatggtgc gctcctggac gtagccttcg ggcatggcgg
     4621 acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg gctgaagcac tgcacgccgt
     4681 aggtcagggt ggtcacgagg gtgggccagg gcacgggcag cttgccggtg gtgcagatga
     4741 acttcagggt cagcttgccg taggtggcat cgccctcgcc ctcgccggac acgctgaact
     4801 tgtggccgtt tacgtcgccg tccagctcga ccaggatggg caccaccccg gtgaacagct
     4861 cctcgccctt gctcaccatg gtggcgaccg gtacccgggg atccaattct ttgccaaaat
     4921 gatgagacag cacaataacc agcacgttgc ccaggagctg taggaaaaag aagaaggcat
     4981 gaacatggtt agcagaggct ctagagccgc cggtcacacg ccagaagccg aaccccgccc
     5041 tgccccgtcc cccccgaagg cagccgtccc cccgcggaca gccccgagcc gctccccccg
     5101 cacaaagggc cctcccggag cccctcaagg ctttcacgca gccacagaaa agaaacaagc
     5161 cgtcattaaa ccaagcgcta attacagccc ggaggagaag ggccgtcccg cccgctcacc
     5221 tgtgggagta acgcggtcag tcagagccgg ggcgggcggc gcgaggcggc ggcggagcgg
     5281 ggcacggggc gaaggcagcg cgcagcgact cccgcccgcc gcgcgcttcg ctttttatag
     5341 ggccgccgcc gccgccgcct cgccataaaa ggaaactttc ggagcgcgcc gctctgattg
     5401 gctgccgccg cacctctccg cctcgccccg ccc
//
