LOCUS       sequence-474414-        5040 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV construct used as a cell filler to label neurons with 
            mTurquoise2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5040)
  TITLE     AAV-constructs used for labeling and synapse inactivation
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5040)
  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            "Unpublished"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-mTurquoise2
FEATURES             Location/Qualifiers
     source          1..5040
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..482
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      528..537
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(534..536,537..539,540..1253)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(579..600)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(840..859)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1187..1208
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1273..1299
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    1325..1913
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1378..1398)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1796..1807)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    1938..2059
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1949..1968)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2003..2022
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2159..2288
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2350..2938)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2430..2449)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3109..3900,3901..3969))
                     /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     3732..3751
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3970..4074)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4142..4160
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4198..4220)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4320..4339
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(4356..4811)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4493..4514)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4705..4724
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   4886..5026
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4897..5026
                     /label=AAV2 ITR
ORIGIN
        1 gcgtgtgtct agactgcaga gggccctgcg tatgagtgca agtgggtttt aggaccagga
       61 tgaggcgggg tgggggtgcc tacctgacga ccgaccccga cccactggac aagcacccaa
      121 cccccattcc ccaaattgcg catcccctat cagagagggg gaggggaaac aggatgcggc
      181 gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt gccttcgccc ccgcctggcg
      241 gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac gtcactcgcc ggtcccccgc
      301 aaactcccct tcccggccac cttggtcgcg tccgcgccgc cgccggccca gccggaccgc
      361 accacgcgag gcgcgagata ggggggcacg ggcgcgacca tctgcgctgc ggcgccggcg
      421 actcagcgct gcctcagtct gcggtgggca gcggaggagt cgtgtcgtgc ctgagagcgc
      481 agggtttagt gaaccgtcag atccgctagc gctaccggac tcagatggcc accatggtga
      541 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
      601 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
      661 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
      721 ccaccctgtc ctggggcgtg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg
      781 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
      841 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
      901 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
      961 agtacaacta ctttagcgac aacgtctata tcaccgccga caagcagaag aacggcatca
     1021 aggccaactt caagatccgc cacaacatcg aggacggcgg cgtgcagctc gccgaccact
     1081 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     1141 gcacccagtc caagctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     1201 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag taaggatggg
     1261 gacagcggcg gataccctta cgacgtgcct gactacgcct gactgcagaa cggccgttat
     1321 cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     1381 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     1441 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     1501 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     1561 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     1621 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     1681 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct
     1741 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     1801 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     1861 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgatt
     1921 gcggccgtca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     1981 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     2041 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     2101 ggagatgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta gcggccgcag
     2161 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
     2221 gggcgaccaa aggtcgcccg acgcccgggc ggcctcagtg agcgagcgag cgcgcagctg
     2281 cctgcaggac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     2341 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     2401 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     2461 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     2521 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     2581 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     2641 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     2701 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     2761 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     2821 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     2881 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     2941 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     3001 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     3061 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     3121 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     3181 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     3241 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     3301 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     3361 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     3421 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     3481 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     3541 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     3601 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     3661 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     3721 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     3781 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     3841 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     3901 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     3961 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     4021 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     4081 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     4141 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     4201 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     4261 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     4321 ggcatcagag cagattgtac tgagagtgca ccataaaatt gtaaacgtta atattttgtt
     4381 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaataga ccgaaatcgg
     4441 caaaatccct tataaatcaa aagaatagcc cgagatagag ttgagtgttg ttccagtttg
     4501 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     4561 tcagggcgat ggcccactac gtgaaccatc acccaaatca agttttttgg ggtcgaggtg
     4621 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     4681 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctaaggcgct
     4741 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     4801 acagggcgcg tactatggtt gctttgacgt atgcggtgtg aaataccgca cagatgcgta
     4861 aggagaaaat accgcatcag gcgcccctgc aggcagctgc gcgctcgctc gctcactgag
     4921 gccgcccggg caaagcccgg gcgtcgggcg acctttggtc gcccggcctc agtgagcgag
     4981 cgagcgcgca gagagggagt ggccaactcc atcactaggg gttcctgcgg ccgcggtacc
//
