LOCUS       sequence-348392-        5770 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses GFP-Cre under the control of a human synapsin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5770)
  TITLE     Widespread inhibitory projections from the cerebellar interposed 
            nucleus
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5770)
  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            "Unpublished"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 hSyn GFP-Cre
FEATURES             Location/Qualifiers
     source          1..5770
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             224..271
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             join(275..277,278..988)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(275..277,278..988)
                     /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(578..597)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     925..946
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1010..2038
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="SNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLL
                     SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP
                     RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL
                     GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER
                     WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR
                     YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED
                     GD"
     polyA_signal    2097..2573
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2296..2315)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2613..2753
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2613..2742
                     /label=AAV2 ITR
     rep_origin      2828..3283
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2915..2934)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3125..3146
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3300..3319)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3419..3441
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3479..3497)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3565..3669
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3670..3738,3739..4530)
                     /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(3888..3907)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4701..5289
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5190..5209
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   5351..5480
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        join(5517..5770,1..194)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
ORIGIN
        1 ccggtccccc gcaaactccc cttcccggcc accttggtcg cgtccgcgcc gccgccggcc
       61 cagccggacc gcaccacgcg aggcgcgaga taggggggca cgggcgcgac catctgcgct
      121 gcggcgccgg cgactcagcg ctgcctcagt ctgcggtggg cagcggagga gtcgtgtcgt
      181 gcctgagagc gcagtctaga ggatcgaatt cacaaccatg gtgaagcgac cagcagcaac
      241 aaagaaggca ggacaagcaa agaagaagaa gctcgtgagc aagggcgagg agctgttcac
      301 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
      361 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
      421 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
      481 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
      541 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
      601 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
      661 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
      721 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
      781 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
      841 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa
      901 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
      961 cactctcggc atggatgagc tgtacaaggg caagaagaag aggaaggtgt ccaatttact
     1021 gaccgtacac caaaatttgc ctgcattacc ggtcgatgca acgagtgatg aggttcgcaa
     1081 gaacctgatg gacatgttca gggatcgcca ggcgttttct gagcatacct ggaaaatgct
     1141 tctgtccgtt tgccggtcgt gggcggcatg gtgcaagttg aataaccgga aatggtttcc
     1201 cgcagaacct gaagatgttc gcgattatct tctatatctt caggcgcgcg gtctggcagt
     1261 aaaaactatc cagcaacatt tgggccagct aaacatgctt catcgtcggt ccgggctgcc
     1321 acgaccaagt gacagcaatg ctgtttcact ggttatgcgg cggatccgaa aagaaaacgt
     1381 tgatgccggt gaacgtgcaa aacaggctct agcgttcgaa cgcactgatt tcgaccaggt
     1441 tcgttcactc atggaaaata gcgatcgctg ccaggatata cgtaatctgg catttctggg
     1501 gattgcttat aacaccctgt tacgtatagc cgaaattgcc aggatcaggg ttaaagatat
     1561 ctcacgtact gacggtggga gaatgttaat ccatattggc agaacgaaaa cgctggttag
     1621 caccgcaggt gtagagaagg cacttagcct gggggtaact aaactggtcg agcgatggat
     1681 ttccgtctct ggtgtagctg atgatccgaa taactacctg ttttgccggg tcagaaaaaa
     1741 tggtgttgcc gcgccatctg ccaccagcca gctatcaact cgcgccctgg aagggatttt
     1801 tgaagcaact catcgattga tttacggcgc taaggatgac tctggtcaga gatacctggc
     1861 ctggtctgga cacagtgccc gtgtcggagc cgcgcgagat atggcccgcg ctggagtttc
     1921 aataccggag atcatgcaag ctggtggctg gaccaatgta aatattgtca tgaactatat
     1981 ccgtaacctg gatagtgaaa caggggcaat ggtgcgcctg ctggaagatg gcgattaggc
     2041 tagctagagt cgacctgcag aagcttgcct cgagcagcgc tgctcgagag atctacgggt
     2101 ggcatccctg tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc
     2161 ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct
     2221 ataatattat ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct
     2281 gtagggcctg cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca
     2341 ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg
     2401 gattccaggc atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt
     2461 tcaccatatt ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc
     2521 ctcccaaatt gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt
     2581 tgtaggtaac cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac
     2641 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc
     2701 gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct
     2761 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa
     2821 ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     2881 gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt
     2941 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     3001 cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt
     3061 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3121 aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt
     3181 gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa
     3241 aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt
     3301 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     3361 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     3421 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     3481 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     3541 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     3601 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     3661 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     3721 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     3781 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     3841 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     3901 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     3961 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     4021 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     4081 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     4141 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     4201 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     4261 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     4321 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     4381 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     4441 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     4501 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     4561 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     4621 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     4681 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     4741 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     4801 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     4861 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     4921 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     4981 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5041 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     5101 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     5161 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     5221 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     5281 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     5341 gctcacatgt cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg
     5401 ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa
     5461 ctccatcact aggggttcct gcggccgcac gctaactgca gagggccctg cgtatgagtg
     5521 caagtgggtt ttaggaccag gatgaggcgg ggtgggggtg cctacctgac gaccgacccc
     5581 gacccactgg acaagcaccc aacccccatt ccccaaattg cgcatcccct atcagagagg
     5641 gggaggggaa acaggatgcg gcgaggcgcg tgcgcactgc cagcttcagc accgcggaca
     5701 gtgccttcgc ccccgcctgg cggcgcgcgc caccgccgcc tcagcactga aggcgcgctg
     5761 acgtcactcg
//
