LOCUS       sequence-79323-e        7034 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Forebrain principal neuron expression of eArchT3,0 with physically 
            uncoupled EGFP fluorophore.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7034)
  TITLE     Ting AAV neural silencing tools
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7034)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pAAV-CaMKIIa-eArchT3.0-P2A-EGFP-WPRE-hGHpA
FEATURES             Location/Qualifiers
     source          1..7034
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        164..1452
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     regulatory      1483..1492
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2338..2394
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             join(2395..2397,2398..2400,2401..3114)
                     /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             join(2395..2397,2398..2400,2401..3111)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2440..2461)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2701..2720)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3048..3069
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3161..3749
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3214..3234)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3632..3643)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3781..4257
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3980..3999)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4297..4437
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4297..4426
                     /label=AAV2 ITR
     rep_origin      4512..4967
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4599..4618)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4809..4830
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4984..5003)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5103..5125
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5163..5181)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5249..5353
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5354..5422,5423..6214)
                     /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(5572..5591)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6385..6973
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6874..6893
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtttaat taacattatg gccttaggtc
      181 acttcatctc catggggttc ttcttctgat tttctagaaa atgagatggg ggtgcagaga
      241 gcttcctcag tgacctgccc agggtcacat cagaaatgtc agagctagaa cttgaactca
      301 gattactaat cttaaattcc atgccttggg ggcatgcaag tacgatatac agaaggagtg
      361 aactcattag ggcagatgac caatgagttt aggaaagaag agtccagggc agggtacatc
      421 tacaccaccc gcccagccct gggtgagtcc agccacgttc acctcattat agttgcctct
      481 ctccagtcct accttgacgg gaagcacaag cagaaactgg gacaggagcc ccaggagacc
      541 aaatcttcat ggtccctctg ggaggatggg tggggagagc tgtggcagag gcctcaggag
      601 gggccctgct gctcagtggt gacagatagg ggtgagaaag cagacagagt cattccgtca
      661 gcattctggg tctgtttggt acttcttctc acgctaaggt ggcggtgtga tatgcacaat
      721 ggctaaaaag cagggagagc tggaaagaaa caaggacaga gacagaggcc aagtcaacca
      781 gaccaattcc cagaggaagc aaagaaacca ttacagagac tacaaggggg aagggaagga
      841 gagatgaatt agcttcccct gtaaacctta gaacccagct gttgccaggg caacggggca
      901 atacctgtct cttcagagga gatgaagttg ccagggtaac tacatcctgt ctttctcaag
      961 gaccatccca gaatgtggca cccactagcc gttaccatag caactgcctc tttgccccac
     1021 ttaatcccat cccgtctgtt aaaagggccc tatagttgga ggtgggggag gtaggaagag
     1081 cgatgatcac ttgtggacta agtttgttcg catccccttc tccaaccccc tcagtacatc
     1141 accctggggg aacagggtcc acttgctcct gggcccacac agtcctgcag tattgtgtat
     1201 ataaggccag ggcaaagagg agcaggtttt aaagtgaaag gcaggcaggt gttggggagg
     1261 cagttaccgg ggcaacggga acagggcgtt tcggaggtgg ttgccatggg gacctggatg
     1321 ctgacgaagg ctcgcgaggc tgtgagcagc cacagtgccc tgctcagaag ccccaagctc
     1381 gtcagtcaag ccggttctcc gtttgcactc aggagcacgg gcaggcgagt ggcccctagt
     1441 tctgggggca gcgggggatc cttcgaaacc ggtgctagcg gcgccaccat ggacccaatt
     1501 gcactgcagg cggggtatga cctcctcggt gacgggaggc ccgagacact ctggctgggc
     1561 atcgggacgc ttctcatgct catcggaacg ttttacttca tcgtcaaagg atggggtgta
     1621 acagacaagg aagcgcgcga gtactactcg atcactattc ttgtgccggg catcgcctca
     1681 gcggcgtatt tgagcatgtt ctttggcatc gggctgaccg aagtgacggt agcgggagag
     1741 gtcttggata tctactatgc gcgctacgca gactggctgt ttacaactcc gctccttttg
     1801 cttgatcttg ctctgttggc aaaagtagat agggtgtcga ttgggacgct ggtgggagta
     1861 gatgccctta tgattgtcac gggattgatt ggggcattgt cgcacacgcc tctcgcccga
     1921 tattcgtggt ggttgttttc caccatctgc atgatcgtgg tcctctactt cttggcgacg
     1981 agccttcggg ctgcggcgaa ggagagagga cccgaggtcg cgtccacatt caacaccttg
     2041 actgccctgg tgctggtgct ctggaccgcg tatcccatct tgtggattat cggaactgaa
     2101 ggtgccgggg tcgtcggtct gggaatcgaa acactcctct tcatggtgtt ggacgtaaca
     2161 gccaaggtcg gatttggttt catcttgctg cggtcaagag caattcttgg ggatacggaa
     2221 gccccggagc ctgcggccgc caagagcagg atcaccagcg agggcgagta catccccctg
     2281 gaccagatcg acatcaacgt gttctgctac gagaacgagg tggttaacgg aagcggagcc
     2341 actaacttct ccctgttgaa acaagcaggg gatgtcgaag agaatcccgg gccaatggtg
     2401 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     2461 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     2521 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     2581 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     2641 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     2701 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     2761 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     2821 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     2881 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     2941 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     3001 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     3061 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaagtcgac
     3121 ggcgcgcccc tgcagggaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa
     3181 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     3241 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     3301 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     3361 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     3421 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     3481 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     3541 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctatgttg ccacctggat
     3601 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     3661 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     3721 tcggatctcc ctttgggccg cctccccgca tcgataccga gcgctgctcg agagatctac
     3781 gggtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca
     3841 gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc
     3901 ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca
     3961 acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg
     4021 ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg
     4081 ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg
     4141 ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct
     4201 tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg
     4261 attttgtagg taaccacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg
     4321 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
     4381 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc
     4441 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa
     4501 gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     4561 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     4621 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     4681 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
     4741 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4801 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cgggctattc
     4861 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     4921 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct
     4981 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     5041 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     5101 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     5161 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     5221 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     5281 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     5341 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     5401 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     5461 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     5521 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     5581 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     5641 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     5701 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     5761 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     5821 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     5881 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     5941 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     6001 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     6061 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     6121 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     6181 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     6241 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     6301 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     6361 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     6421 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     6481 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     6541 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     6601 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     6661 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     6721 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     6781 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     6841 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     6901 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     6961 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     7021 ttttgctcac atgt
//
