LOCUS       sequence-142411-        6854 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An AAV packaging vector that expresses eGFP and Kv1.2 under the 
            control of a neuronal promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6854)
  AUTHORS   Kourrich et al.
  TITLE     AAV Potassium channel
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6854)
  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            "Unpublished"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: pOTTC533 - pAAV SYN1 eGFP-2A-mKv1.2
FEATURES             Location/Qualifiers
     source          1..6854
                     /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        186..633
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      653..662
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(659..661,662..664,665..1375)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(659..661,662..664,665..1375)
                     /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(704..725)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(965..984)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1312..1333
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1397..1450
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     misc_feature    2981..3569
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3034..3054)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3452..3463)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3601..4077
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3800..3819)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4117..4257
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4117..4246
                     /label=AAV2 ITR
     rep_origin      4332..4787
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4419..4438)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4629..4650
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4804..4823)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4923..4945
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4983..5001)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5069..5173
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5174..5242,5243..6034)
                     /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(5392..5411)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6205..6793
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6694..6713
                     /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 cgcgtgtgtc tagactgcag agggccctgc
      181 gtatgagtgc aagtgggttt taggaccagg atgaggcggg gtgggggtgc ctacctgacg
      241 accgaccccg acccactgga caagcaccca acccccattc cccaaattgc gcatccccta
      301 tcagagaggg ggaggggaaa caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca
      361 ccgcggacag tgccttcgcc cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa
      421 ggcgcgctga cgtcactcgc cggtcccccg caaactcccc ttcccggcca ccttggtcgc
      481 gtccgcgccg ccgccggccc agccggaccg caccacgcga ggcgcgagat aggggggcac
      541 gggcgcgacc atctgcgctg cggcgccggc gactcagcgc tgcctcagtc tgcggtgggc
      601 agcggaggag tcgtgtcgtg cctgagagcg cagtcgagaa ggtaccggat ccgccaccat
      661 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
      721 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
      781 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
      841 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
      901 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
      961 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     1021 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     1081 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     1141 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     1201 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     1261 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     1321 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagatgca
     1381 ttctagaggc tccggagagg gcaggggaag tcttctaaca tgcggggacg tggaggaaaa
     1441 tcccgggccc atcgatatga cagtggctac cggagaccca gtggacgagg ctgctgccct
     1501 ccctgggcac ccacaagaca cctatgaccc agaggcagac catgaatgct gtgagagagt
     1561 ggtcatcaac atctcaggcc tgcggttcga aactcagcta aagaccttag cccagttccc
     1621 agagaccctc ttaggggacc ccaagaaacg gatgaggtac ttcgatcccc tccgaaatga
     1681 gtactttttt gatcgcaacc gccctagctt tgatgccatt ttgtactact atcagtctgg
     1741 gggcaggttg aggcgacctg tgaacgtgcc cttagatatc ttctcggaag aaatccggtt
     1801 ttatgagcta ggagaagaag caatggagat gtttcgggag gatgaaggct acatcaagga
     1861 agaagagcgt cctctgcctg aaaatgagtt tcagagacag gtgtggcttc tctttgaata
     1921 ccctgagagc tcagggcctg ccaggattat agccattgta tctgtgatgg tcattctgat
     1981 ctccatcgtc agcttctgtc tggaaacctt gcccatcttc cgggatgaga atgaggacat
     2041 gcatggtggc ggggtgacct tccacaccta ttccaacagc accatcgggt accagcagtc
     2101 cacctccttc accgaccctt tcttcattgt agagactctc tgcatcatct ggttctcctt
     2161 tgagtttctg gttagattct ttgcctgtcc cagcaaagct ggcttcttca ccaacatcat
     2221 gaacatcatt gacattgtgg ctatcatccc ttactttatc accctgggga cagagttagc
     2281 tgagaagcca gaggatgccc agcaaggcca gcaggccatg tcactggcca ttctccgtgt
     2341 catccggttg gtaagagtct ttaggatttt caagttgtcc agacactcca aaggtctaca
     2401 gattctaggt cagaccctca aagctagcat gagggaattg ggcctcctga tattcttcct
     2461 cttcattggg gtcatcctct tctctagtgc tgtctatttt gcagaagctg atgagagaga
     2521 ttcccagttc cccagcatcc cggatgcttt ctggtgggca gtcgtctcca tgacaactgt
     2581 aggctatgga gacatggttc caactaccat tggggggaag atagtgggtt ctctgtgtgc
     2641 aattgcaggt gtgttaacca ttgccttacc agtccctgtc atagtgtcta atttcaacta
     2701 cttctaccac cgggagacag agggagagga gcaggcccag tacttgcaag tgacaagctg
     2761 tccaaagatc ccgtcctccc ctgacctaaa gaaaagtaga agtgcctcta ccataagtaa
     2821 gtctgattac atggagatac aggagggagt taacaacagc aatgaggact ttagagagga
     2881 gaacttaaaa acagccaact gtaccttggc taacacaaac tatgtgaata ttaccaaaat
     2941 gttaactgat gtctgagaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa
     3001 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     3061 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     3121 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     3181 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     3241 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     3301 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     3361 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctatgttg ccacctggat
     3421 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     3481 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     3541 tcggatctcc ctttgggccg cctccccgca tcgataccga gcgctgctcg agagatctac
     3601 gggtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca
     3661 gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc
     3721 ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca
     3781 acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg
     3841 ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg
     3901 ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg
     3961 ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct
     4021 tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg
     4081 attttgtagg taaccacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg
     4141 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
     4201 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc
     4261 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa
     4321 gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     4381 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     4441 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     4501 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
     4561 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4621 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cgggctattc
     4681 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     4741 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct
     4801 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     4861 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     4921 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     4981 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     5041 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     5101 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     5161 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     5221 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     5281 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     5341 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     5401 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     5461 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     5521 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     5581 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     5641 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     5701 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     5761 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     5821 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     5881 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     5941 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     6001 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     6061 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     6121 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     6181 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     6241 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     6301 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     6361 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     6421 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     6481 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     6541 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     6601 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     6661 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     6721 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     6781 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     6841 ttttgctcac atgt
//
