LOCUS       sequence-107238-        7233 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TVA66T-EGFP-B19G under the synapsin promotor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7233)
  TITLE     Genetic Neuroengineering Group Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7233)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pAAV-synP-FLEX-TVA66T-EGFP-B19G
FEATURES             Location/Qualifiers
     source          1..7233
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        164..611
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(620..653)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(704..737)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     CDS             complement(2319..2375)
                     /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             complement(join(2385..3095,3096..3098))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             complement(join(2385..3095,3096..3098))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2427..2448)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2776..2795
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3035..3056
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3533..3542
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    3551..3584
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    3635..3668
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     misc_feature    3674..4262
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3727..3747)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4145..4156)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4270..4494
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4496..4636
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4496..4625
                     /label=AAV2 ITR
     rep_origin      4711..5166
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4798..4817)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5008..5029
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5183..5202)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5302..5324
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5362..5380)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5448..5552
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5553..5621,5622..6413)
                     /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(5771..5790)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6584..7172
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7073..7092
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gtcgacgcta gactgcagag ggccctgcgt atgagtgcaa gtgggtttta
      181 ggaccaggat gaggcggggt gggggtgcct acctgacgac cgaccccgac ccactggaca
      241 agcacccaac ccccattccc caaattgcgc atcccctatc agagaggggg aggggaaaca
      301 ggatgcggcg aggcgcgtgc gcactgccag cttcagcacc gcggacagtg ccttcgcccc
      361 cgcctggcgg cgcgcgccac cgccgcctca gcactgaagg cgcgctgacg tcactcgccg
      421 gtcccccgca aactcccctt cccggccacc ttggtcgcgt ccgcgccgcc gccggcccag
      481 ccggaccgca ccacgcgagg cgcgagatag gggggcacgg gcgcgaccat ctgcgctgcg
      541 gcgccggcga ctcagcgctg cctcagtctg cggtgggcag cggaggagtc gtgtcgtgcc
      601 tgagagcgca gtcgaattca taacttcgta tagcatacat tatacgaagt tattctttgc
      661 accattctaa agaataacag tgataatttc tgggttaagg caaataactt cgtataggat
      721 actttatacg aagttatgct agctcacagt ctggtctcac ccccactctt gtgtgattcc
      781 catgaagata tgatcttccc gctttgggga gtgactgaca cctccctccc tgtccctctg
      841 agattgtgtt gcgtaggttc tgatcgattg actcttctac aacatgtcat caggaaaatt
      901 atcaacatca aggcagtcag ggcccctgca ctcagtaata catacttccc ccagttcggg
      961 agacccaagt caactcctga gacctgattg tgcacatcgg gaaggtgaac ttcaacaaaa
     1021 tcctcagcct cgtcaccgtc cttgaaaacg gtagacgggt ctgccagggg gtgcacaagg
     1081 gggataaccg aggattccaa caactccata tgttgctgga ggagggatga ttgcatctct
     1141 gggattaaga cattgccgtc aggtcctaat attataccat tgaaaaacac cccgttcaca
     1201 tgaggatgac acctcccccc aactcttaaa cacccttttg aagggaggat ctcattccaa
     1261 gttctgactg acttgtagtg agcatcggct tccatcaagg tcttgttgaa tatggtatat
     1321 gcttttccaa acccagggac aagttttctt aaatgactga gacgtctgaa actcactgac
     1381 ttggttgtca tgatggactc tagtgcatcc agacactcct ctctcttcct gaccaactcc
     1441 tctacaacaa ggtgctcaat ttcgtctgag cgaaagtcgt gcaggttcac caacttatcg
     1501 ggagggcacc atttggtttc atttgatgtt tgcatcgaga cccatgttcc atccataagt
     1561 ctaagtccta gaactccaca taacttgagt ttgcatgctc cttttaaaga cttatatagg
     1621 cctctttcat ctacaaagcc gcaagtctca ctccctttgg atgctctctt ccctctacta
     1681 ttggtaaaaa tgtcacaaga catccctagt ctcggattct cgggcatcca aatggtgtaa
     1741 tcgtggttag tggagcagta ggtagaagac accgctactc ctgagcactt cccgctaggg
     1801 aagaccctcg agtgaaggga tctgtcatat gggtccaaat ctgccacact tggagatatg
     1861 ataacgagag actccttggt ggtttttaca gttcgaagcc agcggtagtc agggtacgga
     1921 ttgtgtagag actcttcata tctggggtca ccggccatct tccagttgta cgcggctcta
     1981 catgcatctg gtgttgggcg gaaatgcttt cttttgaacg tggttgtgac ataaccaacg
     2041 aagttagtgt aggtttcagc ctccgtcaca acgcctgtgc aagtgaaccc gttcactttt
     2101 atggctaaga tgtatccaac tttaagttcc atgtaggaga accctgacag gttggtgcat
     2161 ccttcgtcct ccactaccaa attgtttggg cagctgaggt gatgtatgtc aatcggactc
     2221 cagggaccaa gcttgtctgg tatcgtgtaa atagggaatt tcccaaaaca caatggaaaa
     2281 accagaaggg gtacaaacag gagagcctga ggaaccatag gtccagggtt ctcctccacg
     2341 tctccagcct gcttcagcag gctgaagtta gtagctccgc ttcccttgta cagctcgtcc
     2401 atgccgagag tgatcccggc ggcggtcacg aactccagca ggaccatgtg atcgcgcttc
     2461 tcgttggggt ctttgctcag ggcggactgg gtgctcaggt agtggttgtc gggcagcagc
     2521 acggggccgt cgccgatggg ggtgttctgc tggtagtggt cggcgagctg cacgctgccg
     2581 tcctcgatgt tgtggcggat cttgaagttc accttgatgc cgttcttctg cttgtcggcc
     2641 atgatataga cgttgtggct gttgtagttg tactccagct tgtgccccag gatgttgccg
     2701 tcctccttga agtcgatgcc cttcagctcg atgcggttca ccagggtgtc gccctcgaac
     2761 ttcacctcgg cgcgggtctt gtagttgccg tcgtccttga agaagatggt gcgctcctgg
     2821 acgtagcctt cgggcatggc ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag
     2881 cggctgaagc actgcacgcc gtaggtcagg gtggtcacga gggtgggcca gggcacgggc
     2941 agcttgccgg tggtgcagat gaacttcagg gtcagcttgc cgtaggtggc atcgccctcg
     3001 ccctcgccgg acacgctgaa cttgtggccg tttacgtcgc cgtccagctc gaccaggatg
     3061 ggcaccaccc cggtgaacag ctcctcgccc ttgctcaccc cacccccctt ggatgcgctt
     3121 tcaagactga agatgtcaga cctgcttttt gctttggact tcccccatgc agcgatacca
     3181 cccacagcta ccaggcagca caggagcact gcagtgatca gcatccacat gcggccgtga
     3241 ttcctggctg gcagagcacg tccaggagca ggggcagtgg gagcctctgt gccgttgtcc
     3301 gtgggcaccg cggggatcgc gctggtcccg cagccccagg tgtcccgccc gtcgtcgcag
     3361 tcggggtgtc cgtcgcacag ccagtcctgc gggtaacact ccccgtgggc accgggcggc
     3421 tccgagcagc ggaactgacc gggggggcaa cgggacaaag aaccgttacc ggacccgtta
     3481 ccggtcacgt taccgggcag cagcagcagc agcagcgcgg gcagcagccg cgccatggtg
     3541 gcggcgcgcc ataacttcgt ataatgtatg ctatacgaag ttattaagag gtttcatatt
     3601 gctaatagca gctacaatcc agctaccatt ctgcataact tcgtataaag tatcctatac
     3661 gaagttattc cggaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     3721 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     3781 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     3841 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     3901 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     3961 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     4021 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt
     4081 tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     4141 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     4201 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     4261 gcctggatcc tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     4321 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     4381 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     4441 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcaggaa
     4501 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg
     4561 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg
     4621 cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     4681 tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg
     4741 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     4801 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     4861 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     4921 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     4981 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     5041 accctatctc gggctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     5101 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     5161 caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     5221 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     5281 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     5341 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     5401 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     5461 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     5521 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     5581 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     5641 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     5701 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     5761 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     5821 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     5881 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     5941 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     6001 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     6061 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     6121 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     6181 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     6241 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     6301 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     6361 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     6421 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     6481 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     6541 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     6601 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     6661 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     6721 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     6781 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     6841 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     6901 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     6961 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     7021 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     7081 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     7141 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     7201 tcctggcctt ttgctggcct tttgctcaca tgt
//
